AU2019264591B2 - RNA interference mediated inhibition of hepatitis B virus (HBV) gene expression using short interfering nucleic acid (siNA) - Google Patents
RNA interference mediated inhibition of hepatitis B virus (HBV) gene expression using short interfering nucleic acid (siNA) Download PDFInfo
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Abstract
RNA INTERFERENCE MEDIATED INHIBITION OF HEPATITIS B VIRUS (HBV)
GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
Abstract
The present invention relates to compounds, compositions, and methods for the study,
diagnosis, and treatment of traits, diseases and conditions that respond to the modulation of HBV
gene expression and/or activity, and/or modulate a HBV gene expression pathway. Specifically,
the invention relates to double-stranded nucleic acid molecules including small nucleic acid
molecules, such as short interfering nucleic acid (siNA), short interfering RNA (siRNA), double
stranded RNA (dsRNA), micro-RNA (miRNA), and short hairpin RNA (shRNA) molecules that
are capable of mediating or that mediate RNA interference (RNAi) against HBV gene
expression.
AH26(23789354 1):RTK
Description
RNA INTERFERENCE MEDIATED INHIBITION OF HEPATITIS B VIRUS (HBV) GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA)
Cross Reference
[0001] The present application is a divisional application of Australian Patent Application No. 2017216560, the content of which is incorporated herein by reference in its entirety.
[0001a] Australian Patent Application No. 2017216560 is a divisional application of Australian Patent Application No. 2015210336, the content of which is also incorporated herein by reference in its entirety.
[0001b] Australian Patent Application No. 2015210336 is a divisional application of Australian Patent Application No. 20112922261, the content of which is also incorporated herein by reference in its entirety.
[0001c] Australian Patent Application No. 2011292261 is the Australian National Phase Entry for International Patent Application No. PCT/US2011/047512, which claims priority to US 61/374,555, filed on 17 August 2010, the content of which is also incorporated herein by reference in its entirety.
[0001d] The sequence listing submitted via EFS, in compliance with 3 7 CFR §1.52(e)(5), is incorporated herein by reference. The sequence listing text file submitted via EFS contains the file "SequenceListingIFDIUSPSP", created on August 17, 2010, which is 455,495 bytes in size.
[0002] Hepatitis B is an infectious disease of the liver caused by the Hepatitis B virus (HBV). The illness can be acute causing liver inflammation, vomiting, jaundice and in some rare instances of severe fulminant disease, death. The majority of infections result in chronic illness that can be either asymptomatic or resulting in chronic liver inflammation leading to cirrhosis of the liver and an increased incidence in the development of hepatocellular carcinoma (HCC).
[0003] HBV infection is a global problem with approximately > 350 million people world wide chronically infected and 600,000 die each year from HBV-related liver disease or HCC. The disease has caused epidemics in Asia and Africa and is endemic in China. Transmission of HBV is via infectious blood or body fluids. There are currently vaccines for the prevention of HBV
AH26(23789354 1):RTK infection. However, the vaccine is prophylatic and cannot be used to treat already infected patients.
[0004] There are several approved chemotherapeutic treatments for chronic hepatitis B (lamivudine, adefovir, entacavir and telbivudine) that prevent replication of HBV by blocking the action of the HBV polymerase. None of the treatments result in complete clearance of the virus and these treatment result in drug-resistant HBV variants developing after prolonged treatment. Hepatitis B can also be treated with pegylated Interferon-alpha2a but this treatment is associated with severe side effects and is not effective in all patients.
[0005] HBV is a member of the Hepadanvirus family and is divided into 4 major serotyoes (adr, adw, ayr, ayw) based on antigenic epitopes. The virus is also classed into eight genoypes
(A-H based on genomic sequence. Genotype A is common in the Americas, Africa, India and Western Europe. Genotype B and C are found in Asia and the US. Genotype D is most common in Southern Europe and India. Genotype E is found in Western and Southern Africa. Genotype F and H are commonly found in Central and Southern America. Genotype G is commonly found in Europe and the U.S.
[0006] HBV genome consists of a circular strand of DNA that is partially double stranded. The genome is ~3.3Kb in length. It encodes 4 known genes (C, X, P and S). HBV is one of the few DNA viruses that utilize reverse transcriptase in the replication process. HBV replication involves multiples stages including entry, uncoating and transport of the virus genome to the nucleus. Initially, replication of the HBV genome involves the generation of an RNA intermediate that is then reverse transcribed to produce the DNA viral genome.
[0007] Since current therapies are limited due to their ineffectiveness, serious side effects or due to the generation of drug resistant variants, there is a clinical need for the development of new therapies to treat HBV infection.
[0008] Alteration of viral gene expression, specifically HBV gene expression, through RNA interference (hereinafter "RNAi") is one approach for meeting this need. RNAi is induced by short single-stranded RNA ("ssRNA") or double-stranded RNA ("dsRNA") molecules. The short dsRNA molecules, called "short interfering nucleic acids ("siNA")" or "short interfering RNA" or "siRNA" or "RNAi inhibitors" silence the expression of messenger RNAs ("mRNAs") that share sequence homology to the siNA. This can occur via cleavage of the mRNA mediated by an endonuclease complex containing a siNA, commonly referred to as an RNA-induced silencing complex (RISC). Cleavage of the target RNA typically takes place in the middle of the region complementary to the guide sequence of the siNA duplex (Elbashir et al., 2001, Genes Dev., 15:188). In addition, RNA interference can also involve small RNA (e.g., micro-RNA or miRNA) mediated gene silencing, presumably through cellular mechanisms that either inhibit translation or that regulate chromatin structure and thereby prevent transcription of target gene sequences (see for example Allshire, 2002, Science, 297:1818-1819; Volpe et al., 2002, Science, 297:1833-1837; Jenuwein, 2002, Science, 297:2215-2218; and Hall et al., 2002, Science, 297:2232-2237).
[0009] Several studies have attempted to use RNAi for the treatment of HBV and this approach has been comprehensively reviewed in RNAIfor treating Hepatitis B Viral Infection, Chen, Y. et al., PharmaceuticalResearch, 2008 Vol. 25, No.1, pgs 72-86. Yet, as noted in the above reference transfection of a single siRNA often fails to provide adequate gene silencing. Id. Thus, despite significant advances in the field of RNAi, there remains a need for agents that can effectively inhibit HBV gene expression and that can treat disease associated with HBV expression such as liver disease and cancer.
[0010] The invention provides a solution to the problem of treating diseases that respond to the modulation of the HBV gene expression using novel short interfering nucleic acid (siNA) molecules to modulate HBV expression.
[0011] The present invention provides compounds, compositions, and methods useful for modulating the expression of HBV genes and for treating hepatitis B and such conditions that result from hepatitis B infection by RNA interference (RNAi) using small nucleic acid molecules.
[0012] In particular, the instant invention features small nucleic acid molecules, i.e., short interfering nucleic acid (siNA) molecules including, but not limited to, short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), short hairpin RNA (shRNA) and circular RNA molecules and methods used to modulate the expression of HBV genes and/or other genes involved in pathways of HBV gene expression and/or activity. The inventors have found that selection of targets sites from the conserved regions of all known full HBV genomes resulted in sequences with high potency and efficacy.
[0013] In one aspect, the invention provides double-stranded short interfering nucleic acid (siNA) molecules that inhibit the expression of a HBV gene in a cell or mammal, wherein the double-stranded siNAs comprise a sense and an antisense stand. The antisense strand comprises a sequence that is complementary to at least a part of an RNA associated with the expression of the HBV gene. The sense strand comprises a sequence that is complementary to the antisense strand. In various embodiments, at least one strand comprises at least a 15 nucleotide sequence selected from the group of sequences consisting of SEQ ID NOS:1-502. In certain embodiments, the antisense strand comprises at least 15 nucleotides having sequence complementarity to a target sequence set forth in Table la. In other and/or in the same embodiments, the antisense strand comprises at least a 15 nucleotide sequence of one of the antisense sequences set forth in Table 1b. In some embodiments, the sense strand comprises at least a 15 nucleotide sequence of a sense strand sequence as set forth in Table 1b.
[0014] In certain embodiments of this aspect of the invention, double-stranded short interfering nucleic acid (siNA) molecules are provided wherein the antisense stand comprises a modified sequence as set forth in Table le that has sequence complementarity to a target sequence of the invention. In some embodiments, the sense strand also comprises a modified sequence as set forth in Table 1c.
[0015] In certain embodiments, the present invention provides a double-stranded short interfering nucleic acid (siNA) molecule that modulates the expression of HBV, wherein the siNA comprises a sense strand and an antisense strand; each strand is independently 15 to 30 nucleotides in length; and the antisense strand comprises at least 15 nucleotides having sequence complementary to any of:
5'- UCGUGGUGGACUUCUCUCA -3' (SEQ ID NO: 1); 5'- GUGGUGGACUUCUCUCAAU -3' (SEQ ID NO: 2); 5'- GCCGAUCCAUACUGCGGAA -3' (SEQ ID NO: 3); or 5'- CCGAUCCAUACUGCGGAAC -3' (SEQ ID NO: 4).
[0016] In some embodiments of the invention, the antisense strand of a siNA molecule comprises at least a 15 nucleotide sequence of:
5'-UGAGAGAAGUCCACCACGA-3'(SEQ ID NO: 452); 5'-AUUGAGAGAAGUCCACCAC-3'(SEQ ID NO: 453); 5'-UUCCGCAGUAUGGAUCGGC-3'(SEQ ID NO: 454); or 5'-GUUCCGCAGUAUGGAUCGG-3'(SEQ ID NO: 455).
[0017] In some embodiments, the sense strand of a siNA molecule of the invention comprises at least a 15 nucleotide sequence of:
5'- UCGUGGUGGACUUCUCUCA -3' (SEQ ID NO: 1); 5'- GUGGUGGACUUCUCUCAAU -3' (SEQ ID NO: 2); 5'- GCCGAUCCAUACUGCGGAA -3' (SEQ ID NO: 3); or 5'- CCGAUCCAUACUGCGGAAC -3' (SEQ ID NO: 4).
[0018] In some embodiments, a siNA molecule of the invention comprises any of:
5'-UCGUGGUGGACUUCUCUCA-3' (SEQ ID NO: 1) and 5' UGAGAGAAGUCCACCACGA -3'(SEQ ID NO: 452); or 5'-GUGGUGGACUUCUCUCAAU-3' (SEQ ID NO: 2); and 5' AUUGAGAGAAGUCCACCAC -3'(SEQ ID NO: 453); or 5'-GCCGAUCCAUACUGCGGAA-3' (SEQ ID NO: 3) and 5' UUCCGCAGUAUGGAUCGGC -3'(SEQ ID NO: 454); or 5'-CCGAUCCAUACUGCGGAAC-3' (SEQ ID NO: 4); and 5' GUUCCGCAGUAUGGAUCGG -3'(SEQ ID NO: 455).
[0019] In some embodiments, the invention features a double-stranded short interfering nucleic acid (siNA) molecule comprising any of duplex R-008380648-OOOE, R-008380783 OOOR, R-008380665-OOON, R-008380645-000D, R-008380408-OOOR, R-008380633-OOOU, R 008380767-OOOR, R-008380730-OOOC, R-008380777-OOOH, R-008380740-OOOV, R-008380558 000M, R-008380406-000Y, R-008380764-000P, R-008380772-000P, R-008380389-000D, R 008380362-000H, R-008380363-000S, R-008380340-000F, R-008380689-000H, R-008380756 000P, R-008380736-000E, R-008380757-000Y, R-008380753-000N, R-008380737-000N, R 008380734-000M, or R-008380791-000R
[0020] In some embodiments, the invention features a composition comprising:
(a) a double-stranded short interfering nucleic acid (siNA) of the invention;
(b) a cationic lipid compond having any of compound numbers 1-46 or any combination thereof;
(c) cholesterol;
(d) DSPC; and
(e) PEG-DMG.
[0021] In some embodiments, a composition of the invention comprises any Cationic Lipid having any of compound numbers 1-46 in the following molar ratios:
Cationic Lipid / Cholesterol / PEG-DMG 56.6/38/5.4; Cationic Lipid / Cholesterol / PEG-DMG 60/38/2; Cationic Lipid/ Cholesterol /PEG-DMG 67.3/29/3.7; Cationic Lipid / Cholesterol /PEG-DMG 49.3/47/3.7; Cationic Lipid / Cholesterol /PEG-DMG 50.3/44.3/5.4; Cationic Lipid / Cholesterol /PEG-C-DMA / DSPC 40/48/2/10; Cationic Lipid / Cholesterol /PEG-DMG / DSPC 40/48/2/10; and Cationic Lipid / Cholesterol /PEG-DMG / DSPC 58/30/2/10.
[0022] In some embodiments, a composition of the invention comprises (13Z,16Z)-N,N dimethyl-3-nonyldocosa-13,16-dien-1-amine, cholesterol, DSPC, and PEG-DMG, having a molar ratio of about 50:30:10:2 respectively.
[0023] In some embodiments, a composition of the invention further comprises a cryo protectant. In some embodiments, the cryoprotectant is Sucrose, Trehalose, Raffinose, Stachyose, Verbascose, Mannitol, Glucose, Lactose, Maltose, Maltotriose-heptaose, Dextran, Hydroxyethyl Starch, Insulin, Sorbitol, Glycerol, Arginine, Histidine, Lysine, Proline, Dimethylsulfoxide or any combination thereof. In some embodiments, the cryoprotectant is Sucrose. In some embodiments, the cryoprotectant is Trehalose. In some embodiments, the cryoprotectant is a combination of Sucrose and Trehalose.
[0024] In some embodiments of the invention, all of the nucleotides of siNAs of the invention are unmodified. In other embodiments, one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) of the nucleotide positions independently in either one or both strands of an siNA molecule are modified.
Modifications include nucleic acid sugar modifications, base modifications, backbone (internucleotide linkage) modifications, non-nucleotide modifications, and/or any combination thereof. In certain instances, purine and pyrimidine nucleotides are differentially modified. For example, purine and pyrimidine nucleotides can be differentially modified at the 2'-sugar position (i.e., at least one purine has a different modification from at least one pyrimidine in the same or different strand at the 2'-sugar position). In certain instances the purines are unmodified in one or both strands, while the pyrimidines in one or both strands are modified. In certain other instances, the pyrimidines are unmodified in one or both strands, while the purines in one or both strands are modified. In some instances, at least one modified nucleotide is a 2'-deoxy-2'-fluoro nucleotide, a 2'-deoxy nucleotide, or a 2'-O-alkyl nucleotide. In some instances, at least 5 or more of the pyrimidine nucleotides in one or both stands are either all 2'-deoxy-2'-fluoro or all 2'-O-methyl pyrimidine nucleotides. In some instances, at least 5 or more of the purine nucleotides in one or both stands are either all 2'-deoxy-2'-fluoro or all 2'--methyl purine nucleotides. In certain instances, wherein the siNA molecules comprise one or more modifications as described herein, the nucleotides at positions 1, 2, and 3 at the 5' end of the guide (antisense) strand are unmodified.
[0025] In certain embodiments, the siNA molecules of the invention have 3' overhangs of one, two, three, or four nucleotide(s) on one or both of the strands. In other embodiments, the siNA molecules lack overhangs (i.e., have blunt ends). Preferably, the siNA molecule has 3' overhangs of two nucleotides on both the sense and antisense strands. The overhangs can be modified or unmodified. Examples of modified nucleotides in the overhangs include, but are not limited to, 2'-O-alkyl nucleotides, 2'-deoxy-2'-fluoro nucleotides, locked nucleic acid (LNA) nucleotides, or 2'-deoxy nucleotides. The overhang nucleotides in the antisense strand can comprise nucleotides that are complementary to nucleotides in the HBV target sequence. Likewise, the overhangs in the sense stand can comprise nucleotides that are in the HBV target sequence. In certain instances, the siNA molecules of the invention have two 3' overhang nucleotides on the antisense stand that are 2'-O-alkyl (e.g., 2'-O-methyl) nucleotides and two 3' overhang nucleotides on the sense stand that are 2'-deoxy nucleotides. In other instances, the siNA molecules of the invention have two 3' overhang nucleotides that are 2'-O-alkyl (e.g., 2'-0 methyl) nucleotides on both the antisense stand and on the sense stand. In certain embodiments, the 2'-O-alkyl nucleotides are 2'-O-methyl uridine nucleotides. In certain instances, the overhangs also comprise one or more phosphorothioate linkages between nucleotides of the overhang.
[0026] In some embodiments, the siNA molecules of the invention have caps (also referred to herein as "terminal caps." The cap can be present at the 5'-terminus (5'-cap) or at the 3' terminus (3'-cap) or can be present on both termini, such as at the 5' and 3'termini of the sense strand of the siNA.
[0027] In some embodiments, the siNA molecules of the invention are phosphorylated at the ' end of the antisense strand. The phosphate group can be a phosphate, a diphosphate or a triphosphate.
[0028] The siNA molecules of the invention when double stranded can be symmetric or asymmetric. Each strand of these double stranded siNAs independently can range in nucleotide length between 3 and 30 nucleotides. Generally, each strand of the siNA molecules of the invention is about 15 to 30 (i.e., about 19, 20, 21, 22, 23 or 24) nucleotides in length.
[0029] The siNA molecules of the invention, which are double stranded or have a duplex structure, independently comprise about 3 to about 30 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) base pairs. Generally, the duplex structure of siNAs of the invention is between 15 and 30, more generally between 18 and , yet more generally between 19 and 24, and most generally between 19 and 21 base pairs in length.
[0030] In certain embodiments, double-stranded short interfering nucleic acid (siNA) molecules are provided, wherein the molecule has a sense strand and an antisense strand and comprises formula (A):
B Nx 3 (N)x2 B -3' B (N)xi Nx 4 [N]x5 -5' (A) wherein, the upper strand is the sense strand and the lower strand is the antisense strand of the double-stranded nucleic acid molecule; wherein the antisense strand comprises at least a 15 nucleotide sequence of SEQ ID NO: 452, SEQ ID NO: 453, SEQ ID NO: 454, or SEQ ID NO: 455, and the sense strand comprises a sequence having complementarity to the antisense strand; each N is independently a nucleotide which is unmodified or chemically modified or a non-nucleotide; each B is a terminal cap that is present or absent;
(N) represents overhanging nucleotides, each of which is independently unmodified or chemically modified;
[N] represents nucleotides that are ribonucleotides;
X1 and X2 are independently integers from 0 to 4;
X3 is an integer from 15 to 30;
X4 is an integer from 9 to 30; and
X5 is an integer from 0 to 6, provided that the sum of X4 and X5 is 15-30.
[0031] In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) of formula (A); wherein (a) one or more pyrimidine nucleotides in Nx 4 positions are independently 2'-deoxy 2'-fluoro nucleotides, 2'-O-alkyl nucleotides, 2'-deoxy nucleotides, ribonucleotides, or any combination thereof;
(b) one or more purine nucleotides in Nx4 positions are independently 2'-deoxy-2' fluoro nucleotides, 2'-O-alkyl nucleotides, 2'-deoxy nucleotides, ribonucleotides, or any combination thereof;
(c) one or more pyrimidine nucleotides in Nx 3 positions are independently 2'-deoxy 2'-fluoro nucleotides, 2'-O-alkyl nucleotides, 2'-deoxy nucleotides, ribonucleotides, or any combination thereof; and
(d) one or more purine nucleotides in Nx3 positions are independently 2'-deoxy-2' fluoro nucleotides, 2'-O-alkyl nucleotides, 2'-deoxy nucleotides, ribonucleotides,
[0032] The present invention further provides compositions comprising the double-stranded nucleic acid molecules described herein with optionally a pharmaceutically acceptable carrier or diluent.
[0033] The administration of the composition can be carried out by known methods, wherein the nucleic acid is introduced into a desired target cell in vitro or in vivo.
[0034] Commonly used techniques for introduction of the nucleic acid molecules of the invention into cells, tissues, and organisms include the use of various carrier systems, reagents and vectors. Non-limiting examples of such carrier systems suitable for use in the present invention include conjugates, nucleic-acid-lipid particles, lipid nanoparticles (LNP), liposomes, lipoplexes, micelles, virosomes, virus like particles (VLP), nucleic acid complexes, and mixtures thereof.
[0035] The compositions of the invention can be in the form of an aerosol, dispersion, solution (e.g., an injectable solution), a cream, ointment, tablet, powder, suspension or the like. These compositions may be administered in any suitable way, e.g. orally, sublingually, buccally, parenterally, nasally, or topically. In some embodiments, the compositions are aerosolized and delivered via inhalation.
[0036] The molecules and compositions of the present invention have utility over a broad range of therapeutic applications. Accordingly another aspect of this invention relates to the use of the compounds and compositions of the invention in treating a subject. The invention thus provides a method for treating a subject, such as a human, suffering from a condition which is mediated by the action of HBV, wherein the method comprises administering to the subject an effective amount of a double-stranded short interfering nucleic acid (siNA) molecule of the invention. Thus, the siNA molecules of the invention treat the hepatitis B infection and/or conditions resulting therefrom. In some embodiments, the condition is liver disease. In other embodiments, the condition is cancer.
[0037] These and other aspects of the invention will be apparent upon reference to the following detailed description and attached figures. Moreover, it is contemplated that any method or composition described herein can be implemented with respect to any other method or composition described herein and that different embodiments may be combined.
[00381 Additionally, patents, patent applications, and other documents are cited throughout the specification to describe and more specifically set forth various aspects of this invention. Each of these references cited herein is hereby incorporated by reference in its entirety, including the drawings.
[0039] Figure 1 shows a non-limiting proposed mechanistic representation of target RNA degradation involved in RNAi. Double-stranded RNA (dsRNA), which is generated by RNA dependent RNA polymerase (RdRP) from foreign single-stranded RNA, for example viral, transposon, or other exogenous RNA, activates the DICER enzyme that in turn generates siNA duplexes. Alternately, synthetic or expressed siNA can be introduced directly into a cell by appropriate means. An active siNA complex forms that recognizes a target RNA, resulting in degradation of the target RNA by the RISC endonuclease complex or in the synthesis of additional RNA by RNA-dependent RNA polymerase (RdRP), which can activate DICER and result in additional siNA molecules, thereby amplifying the RNAi response.
[0040] Figure 2 shows non-limiting examples of chemically modified siNA constructs of the present invention using a generalized structure of a representative siNA duplex. The specific modifications shown in the figure can be utilized alone or in combination with other modifications of the figure, in addition to other modifications and features described herein with reference to any siNA molecule of the invention. In the figure, N stands for any nucleotide or optionally a non-nucleotide as described here. The upper strand, having B-Nx 3-(N)x 2-B -3' is the sense (or passenger) strand of the siNA, whereas the lower strand, having B(N)xi-Nx 4 -[N]x 5 -5' is the antisense (or guide) strand of the siNA. Nucleotides (or optional non-nucleotides) of internal portions of the sense strand are designated Nx3 and nucleotides (or optional non nucleotides) of internal portions of the antisense strand are designated Nx 4 . Nucleotides (or optional non-nucleotides) of the internal portions are generally base paired between the two strands, but can optionally lack base pairing (e.g. have mismatches or gaps) in some embodiments. Nucleotides (or optional non-nucleotides) of overhang regions are designated by parenthesis (N). Nucleotides of the 5'-terminal portion of the antisense strand are designated
[N]. Terminal caps are optionally present at the 5' and/or 3' end of the sense strand and further optionally present at the 3'-end of the antisense strand. Generally, each strand can independently range from about 15 to about 30 nucleotides in length, but can vary depending on the presence of any overhang nucleotides. In certain embodiments, X1 and X2 are independently integers from to 4; X3 is an integer from 15 to 30; X4 is an integer from 9 to 30; X5 is an integer from 0 to 6, provided that the sum of X4 and X5 is 15-30. Various modifications are shown for the nucleotides of the sense and antisense strands of the siNA constructs. The (N) overhang nucleotide positions can be chemically modified as described herein (e.g., 2'-O-methyl, 2' deoxy-2'-fluoro, 2'-deoxy, LNA, universal bases etc.) and can be either derived from a corresponding target nucleic acid sequence or not. The constructs shown in the figure can also comprise phosphorothioate linkages as described herein. For example, phosphorothioate linkages can exist between any N, (N), and/or [N] positions. Such phosphorothioate incorporation can be utilized between purine "R" and pyrimidine "Y" positions, or for stabilization of pyrimidine linkages in general. Furthermore, although not depicted on the Figure, the constructs shown in the figure can optionally include a ribonucleotide at the 9th
position from the 5'-end of the sense strand or the 11" position based on the 5'-end of the guide strand by counting 11 nucleotide positions in from the 5'-terminus of the guide strand. Similarly, the antisense strand can include a ribonucleotide at the 14" position from the 5'-end, or alternately can be selected or designed so that a 2'-O-alkyl nucleotide (e.g., a 2'-O-methyl purine) is not present at this position. Furthermore, although not shown in the Figure, the 5'-terminal position of the antisense strand can comprise a terminal phosphate group as described herein. The antisense strand generally comprises sequence complementary to any target nucleic acid sequence of the invention, such as those set forth in Table la herein.
[0041] Figure 3 shows non-limiting examples of certain combinations of modifications applied to the representative siNA duplex described in Figure 2. The table shown below the representative structure provides specific combinations of (N)x1 , (N)x 2 , Nx 3, Nx 4 , and/or [N]x 5 nucleotide (and optional non-nucleotide) positions. For example, combinations of 5 or more (e.g., 5, 6, 7, 8, 9, or 10 or more) Nx3 and 5 or more (e.g., 5, 6, 7, 8, 9, or 10 or more) Nx 4 pyrimidine "Y" and purine "R" nucleotides are specified, each of which can independently have specific (N)x 1, and/or (N)X2, substitutions as shown in the figure, in addition to optional phosphorothioate substitutions. The 5'-terminal antisense strand [N] nucleotides are generally ribonucleotides, but can also be modified or unmodified depending on if they are purine "R" or pyrimidine "Y" nucleotides
[0042] Figure 4A-C shows non-limiting examples of different siNA constructs of the invention. The criteria of the representative structures shown in Figures 2 and 3 can be applied to any of the structures shown in Figure 4A-C.
[0043] The examples shown in Figure 4A (constructs 1, 2, and 3) have 19 representative base pairs; however, different embodiments of the invention include any number of base pairs described herein. Bracketed regions represent nucleotide overhangs, for example, comprising about 1, 2, 3, or 4 nucleotides in length, preferably about 2 nucleotides. Constructs 1 and 2 can be used independently for RNAi activity. Construct 2 can comprise a polynucleotide or non nucleotide linker, which can optionally be designed as a biodegradable linker. In one embodiment, the loop structure shown in construct 2 can comprise a biodegradable linker that results in the formation of construct 1 in vivo and/or in vitro. In another example, construct 3 can be used to generate construct 2 under the same principle wherein a linker is used to generate the active siNA construct 2 in vivo and/or in vitro, which can optionally utilize another biodegradable linker to generate the active siNA construct 1 in vivo and/or in vitro. As such, the stability and/or activity of the siNA constructs can be modulated based on the design of the siNA construct for use in vivo or in vitro and/or in vitro.
[0044] The examples shown in Figure 4B represent different variations of double-stranded nucleic acid molecule of the invention, such as microRNA, that can include overhangs, bulges, loops, and stem-loops resulting from partial complementarity. Such motifs having bulges, loops, and stem-loops are generally characteristics of miRNA. The bulges, loops, and stem-loops can result from any degree of partial complementarity, such as mismatches or bulges of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more nucleotides in one or both strands of the double-stranded nucleic acid molecule of the invention.
[0045] The example shown in Figure 4C represents a model double-stranded nucleic acid molecule of the invention comprising a 19 base pair duplex of two 21 nucleotide sequences having dinucleotide 3'-overhangs. The top strand (1) represents the sense strand (passenger strand), the middle strand (2) represents the antisense (guide strand), and the lower strand (3) represents a target polynucleotide sequence. The dinucleotide overhangs (NN) can comprise a sequence derived from the target polynucleotide. For example, the 3'-(NN) sequence in the guide strand can be complementary to the 5'-[NN] sequence of the target polynucleotide. In addition, the 5'-(NN) sequence of the passenger strand can comprise the same sequence as the '-[NN] sequence of the target polynucleotide sequence. In other embodiments, the overhangs (NN) are not derived from the target polynucleotide sequence, for example where the 3'-(NN) sequence in the guide strand are not complementary to the 5'-[NN] sequence of the target polynucleotide and the 5'-(NN) sequence of the passenger strand can comprise different sequence from the 5'-[NN] sequence of the target polynucleotide sequence. In additional embodiments, any (NN) nucleotides are chemically modified, e.g., as 2'-0-methyl, 2'-deoxy-2' fluoro, and/or other modifications herein. Furthermore, the passenger strand can comprise a ribonucleotide position N of the passenger strand. For the representative 19 base pair 21 mer duplex shown, position N can be 9 nucleotides in from the 3' end of the passenger strand. However, in duplexes of differing length, the position N is determined based on the 5'-end of the guide strand by counting 11 nucleotide positions in from the 5'-terminus of the guide strand and picking the corresponding base paired nucleotide in the passenger strand. Cleavage by Ago2 takes place between positions 10 and 11 as indicated by the arrow. In additional embodiments, there are two ribonucleotides, NN, at positions 10 and 11 based on the 5'-end of the guide strand by counting 10 and 11 nucleotide positions in from the 5'-terminus of the guide strand and picking the corresponding base paired nucleotides in the passenger strand.
[0046] Figure 5 shows non-limiting examples of different stabilization chemistries (1-10) that can be used, for example, to stabilize the 5' and/or 3'-ends of siNA sequences of the invention, including (1) [3-3']-inverted deoxyribose; (2) deoxyribonucleotide; (3) [5'-3']-3' deoxyribonucleotide; (4) [5'-3']-ribonucleotide; (5) [5'-3']-3'-0-methyl ribonucleotide; (6) 3' glyceryl; (7) [3'-5']-3'-deoxyribonucleotide; (8) [3'-3']-deoxyribonucleotide; (9) [5'-2'] deoxyribonucleotide; and (10) [5-3']-dideoxyribonucleotide (when X = 0). In addition to modified and unmodified backbone chemistries indicated in the figure, these chemistries can be combined with different sugar and base nucleotide modifications as described herein.
[0047] Figure 6 shows a non-limiting example of a strategy used to identify chemically modified siNA constructs of the invention that are nuclease resistant while preserving the ability to mediate RNAi activity. Chemical modifications are introduced into the siNA construct based on educated design parameters (e.g. introducing 2'-modifications, base modifications, backbone modifications, terminal cap modifications etc). The modified construct is tested in an appropriate system (e.g., human serum for nuclease resistance, shown, or an animal model for PK/delivery parameters). In parallel, the siNA construct is tested for RNAi activity, for example in a cell culture system such as a luciferase reporter assay and/or against endogenous mRNA). Lead siNA constructs are then identified which possess a particular characteristic while maintaining RNAi activity, and can be further modified and assayed once again. This same approach can be used to identify siNA-conjugate molecules with improved pharmacokinetic profiles, delivery, and RNAi activity.
[0048] Figure 7 shows non-limiting examples of phosphorylated siNA molecules of the invention, including linear and duplex constructs and asymmetric derivatives thereof.
[0049] Figure 8 shows non-limiting examples of chemically modified terminal phosphate groups of the invention.
[0050] Figure 9 shows a non-limiting example of a cholesterol linked phosphoramidite that can be used to synthesize cholesterol conjugated siNA molecules of the invention. An example is shown with the cholesterol moiety linked to the 5'-end of the sense strand of an siNA molecule.
[0051] Figure 10 depicts an embodiment of 5' and 3' inverted abasic cap linked to a nucleic acid strand.
A. Terms and Definitions
[0052] The following terminology and definitions apply as used in the present application.
[0053] The term "abasic" as used herein refers to its meaning as is generally accepted in the art. The term generally refers to sugar moieties lacking a nucleobase or having a hydrogen atom
(H) or other non-nucleobase chemical groups in place of a nucleobase at the ' position of the sugar moiety, see for example Adamic et al., U.S. Pat. No. 5,998,203. In one embodiment, an abasic moiety of the invention is a ribose, deoxyribose, or dideoxyribose sugar.
[0054] The term "acyclic nucleotide" as used herein refers to its meaning as is generally accepted in the art. The term generally refers to any nucleotide having an acyclic ribose sugar, for example where any of the ribose carbon/carbon or carbon/oxygen bonds are independently or in combination absent from the nucleotide.
[0055] The term "alkyl" as used herein refers to its meaning as is generally accepted in the art. The term generally refers to a saturated or unsaturated hydrocarbons, including straight chain, branched-chain, alkenyl, alkynyl groups and cyclic groups, but excludes aromatic groups. Notwithstanding the foregoing, alkyl also refers to non-aromatic heterocyclic groups. Preferably, the alkyl group has 1 to 12 carbons. More preferably, it is a lower alkyl of from 1 to 7 carbons, more preferably 1 to 4 carbons. The alkyl group can be substituted or unsubstituted. When substituted, the substituted group(s) is preferably, hydroxyl, halogen, cyano, C1-C4 alkoxy, =0, =S, NO 2, SH, NH 2 , or NRR 2 , where R1 and R2 independently are H or Cl-C4 alkyl.
[0056] The phrase "agents that interfere with cell cycle checkpoints" refers to compounds that inhibit protein kinases that transduce cell cycle checkpoint signals, thereby sensitizing the cancer cell to DNA damaging agents.
[0057] The phrase "agents that interfere with receptor tyrosine kinases (RTKs)" refers to compounds that inhibit RTKs and therefore inhibit mechanisms involved in oncogenesis and tumor progression.
[0058] The phrase "androgen receptor modulators" refers to compounds that interfere or inhibit the binding of androgens to the receptor, regardless of mechanism.
[0059] The phrase "angiogenesis inhibitors" refers to compounds that inhibit the formation of new blood vessels, regardless of mechanism.
[0060] The term "aryl" as used herein refers to its meaning as is generally accepted in the art. The term generally refers to an aromatic group that has at least one ring having a conjugated pi electron system and includes carbocyclic aryl, heterocyclic aryl and biaryl groups, all of which can be optionally substituted. The preferred substituent(s) of aryl groups are halogen, trihalomethyl, hydroxyl, SH, OH, cyano, Cl-C4 alkoxy, Cl-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, NH 2 , and NR 1R2 groups, where R 1 and R2 independently are H or Cl-C4 alkyl.
[0061] The term "alkylaryl" as used herein refers to its meaning as is generally accepted in the art. The term generally refers to an alkyl group (as described above) covalently joined to an aryl group (as described above). Carbocyclic aryl groups are groups wherein the ring atoms on the aromatic ring are all carbon atoms. The carbon atoms are optionally substituted. Heterocyclic aryl groups are groups having from 1 to 3 heteroatoms as ring atoms in the aromatic ring and the remainder of the ring atoms are carbon atoms. Suitable heteroatoms include oxygen, sulfur, and nitrogen, and examples of heterocyclic aryl groups having such heteroatoms include furanyl, thienyl, pyridyl, pyrrolyl, N-lower alkyl pyrrolo, pyrimidyl, pyrazinyl, imidazolyl and the like, all optionally substituted. Preferably, the alkyl group is a C1-C4 alkyl group.
[0062] The term "amide" as used herein refers to its meaning as is generally accepted in the art. The term generally refers to an -C(O)-NH-R, where R is either alkyl, aryl, alkylaryl or hydrogen.
[0063] The phrase "antisense region" as used herein refers to its meaning as is generally accepted in the art. With reference to exemplary nucleic acid molecules of the invention, the term refers to a nucleotide sequence of an siNA molecule having complementarity to a target nucleic acid sequence. In addition, the antisense region of an siNA molecule can optionally comprise a nucleic acid sequence having complementarity to a sense region of the siNA molecule. In one embodiment, the antisense region of the siNA molecule is referred to as the antisense strand or guide strand.
[0064] The phrase "asymmetric hairpin" refers to a linear siNA molecule comprising an antisense region, a loop portion that can comprise nucleotides or non-nucleotides, and a sense region that comprises fewer nucleotides than the antisense region to the extent that the sense region has enough complementary nucleotides to base pair with the antisense region and form a duplex with loop. For example, an asymmetric hairpin siNA molecule of the invention can comprise an antisense region having length sufficient to mediate RNAi in a cell or in vitro system (e.g. about 15 to about 30, or about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides) and a loop region comprising about 4 to about 12 (e.g., about 4, 5, 6, 7, 8, 9, 10, 11, or 12) nucleotides, and a sense region having about 3 to about 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) nucleotides that are complementary to the antisense region. The asymmetric hairpin siNA molecule can also comprise a 5'-terminal phosphate group that can be chemically modified. The loop portion of the asymmetric hairpin siNA molecule can comprise nucleotides, non-nucleotides, linker molecules, or conjugate molecules as described herein.
[0065] The term "biodegradable" as used herein refers to its meaning as is generally accepted in the art. The term generally refers to degradation in a biological system, for example, enzymatic degradation or chemical degradation.
[0066] The term "biodegradable linker" as used herein refers to its meaning as is generally accepted in the art. With reference to exemplary nucleic acid molecules of the invention, the term refers to a linker molecule that is designed to connect one molecule to another molecule, and which is susceptible to degradation in a biological system. The linker can be a nucleic acid or non-nucleic acid based linker. For example, a biodegradable linker can be used to attach a ligand or biologically active molecule to an siNA molecule of the invention. Alternately, a biodegradable linker can be used to connect the sense and antisense strands of an siNA molecule of the invention. The biodegradable linker is designed such that its stability can be modulated for a particular purpose, such as delivery to a particular tissue or cell type. The stability of a nucleic acid-based biodegradable linker molecule can be modulated by using various chemistries, for example combinations of ribonucleotides, deoxyribonucleotides, and chemically modified nucleotides, such as 2'-0-methyl, 2'-fluoro, 2'-amino, 2'-0-amino, 2'-C-allyl, 2'-0-allyl, and other 2'-modified or base modified nucleotides. The biodegradable nucleic acid linker molecule can be a dimer, trimer, tetramer or longer nucleic acid molecule, for example, an oligonucleotide of about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleotides in length, or can comprise a single nucleotide with a phosphorus-based linkage, for example, a phosphoramidate or phosphodiester linkage. The biodegradable nucleic acid linker molecule can also comprise nucleic acid backbone, nucleic acid sugar, or nucleic acid base modifications.
[0067] The phrase "biologically active molecule" as used herein refers to its meaning as is generally accepted in the art. With reference to exemplary nucleic acid molecules of the invention, the term refers to compounds or molecules that are capable of eliciting or modifying a biological response in a system and/or are capable of modulating the pharmacokinetics and/or pharmacodynamics of other biologically active molecules. Examples of biologically active molecules, include siNA molecules alone or in combination with other molecules including, but not limited to therapeutically active molecules such as antibodies, cholesterol, hormones, antivirals, peptides, proteins, chemotherapeutics, small molecules, vitamins, co-factors, nucleosides, nucleotides, oligonucleotides, enzymatic nucleic acids, antisense nucleic acids, triplex forming oligonucleotides, polyamines, polyamides, polyethylene glycol, other polyethers, 2-5A chimeras, siNA, dsRNA, allozymes, aptamers, decoys and analogs thereof.
[0068] The phrase "biological system" as used herein refers to its meaning as is generally accepted in the art. The term generally refers to material, in a purified or unpurified form, from biological sources including, but not limited to, human or animal, wherein the system comprises the components required for RNAi activity. Thus, the phrase includes, for example, a cell, tissue, subject, or organism, or extract thereof. The term also includes reconstituted material from a biological source.
[0069] The phrase "blunt end" as used herein refers to its meaning as is generally accepted in the art. With reference to exemplary nucleic acid molecules of the invention, the term refers to termini of a double-stranded siNA molecule having no overhanging nucleotides. For example, the two strands of a double-stranded siNA molecule having blunt ends align with each other with matched base-pairs without overhanging nucleotides at the termini. A siNA duplex molecule of the invention can comprise blunt ends at one or both termini of the duplex, such as termini located at the 5'-end of the antisense strand, the 5'-end of the sense strand, or both termini of the duplex.
[0070] The term "cap" also referred to herein as "terminal cap," as used herein refers to its meaning as is generally accepted in the art. With reference to exemplary nucleic acid molecules of the invention, the term refers to a moiety, which can be a chemically modified nucleotide or non-nucleotide that can be incorporated at one or more termini of one or more nucleic acid molecules of the invention. These terminal modifications protect the nucleic acid molecule from exonuclease degradation, and can help in delivery and/or localization within a cell. The cap can be present at the 5'-terminus (5'-cap) or at the 3'-terminal (3'-cap) or can be present on both termini of any nucleic acid molecule of the invention. A cap can be present at the 5'-end, 3-end and/or 5' and 3'-ends of the sense strand of a nucleic acid molecule of the invention. Additionally, a cap can optionally be present at the 3'-end of the antisense strand of a nucleic acid molecule of the invention. In non-limiting examples, the 5'-cap includes, but is not limited to, LNA; glyceryl; inverted deoxy abasic residue (moiety); 4',5'-methylene nucleotide; 1-(beta D-erythrofuranosyl) nucleotide, 4'-thio nucleotide; carbocyclic nucleotide; 1,5-anhydrohexitol nucleotide; L-nucleotides; alpha-nucleotides; modified base nucleotide; phosphorodithioate linkage; threo-pentofuranosyl nucleotide; acyclic 3',4'-seco nucleotide; acyclic 3,4 dihydroxybutyl nucleotide; acyclic 3,5-dihydroxypentyl nucleotide; 3'-3'-inverted nucleotide moiety; 3'-3'-inverted abasic moiety; 3'-2'-inverted nucleotide moiety; 3'-2'-inverted abasic moiety; 1,4-butanediol phosphate; 3'-phosphoramidate; hexylphosphate; aminohexyl phosphate; 3'-phosphate; 3'-phosphorothioate; phosphorodithioate; or bridging or non-bridging methylphosphonate moiety. Non-limiting examples of the 3'-cap include, but are not limited to, LNA; glyceryl; inverted deoxy abasic residue (moiety); 4', 5'-methylene nucleotide; 1-(beta-D erythrofuranosyl) nucleotide; 4'-thio nucleotide; carbocyclic nucleotide; 5'-amino-alkyl phosphate; 1,3-diamino-2-propyl phosphate; 3-aminopropyl phosphate; 6-aminohexyl phosphate; 1,2-aminododecyl phosphate; hydroxypropyl phosphate; 1,5-anhydrohexitol nucleotide; L nucleotide; alpha-nucleotide; modified base nucleotide; phosphorodithioate; threo pentofuranosyl nucleotide; acyclic 3',4'-seco nucleotide; 3,4-dihydroxybutyl nucleotide; 3,5 dihydroxypentyl nucleotide, 5'-5'-inverted nucleotide moiety; 5'-5'-inverted abasic moiety; 5' phosphoramidate; 5'-phosphorothioate; 1,4-butanediol phosphate; 5'-amino; bridging and/or non bridging 5'-phosphoramidate; phosphorothioate and/or phosphorodithioate; bridging or non bridging methylphosphonate; and 5'-mercapto moieties (for more details see Beaucage and Iyer, 1993, Tetrahedron 49, 1925; incorporated by reference herein). Figure 5 shows some non limiting examples of various caps.
[0071] The term "cell" as used herein refers to its meaning as is generally accepted in the art. With reference to exemplary nucleic acid molecules of the invention, the term is used in its usual biological sense, and does not refer to an entire multicellular organism, e.g., specifically does not refer to a human being. The cell can be present in an organism, e.g., birds, plants and mammals, such as humans, cows, sheep, apes, monkeys, swine, dogs, and cats. The cell can be prokaryotic (e.g., bacterial cell) or eukaryotic (e.g., mammalian or plant cell). The cell can be of somatic or germ line origin, totipotent or pluripotent, dividing or non-dividing. The cell can also be derived from or can comprise a gamete or embryo, a stem cell, or a fully differentiated cell.
[0072] The phrase "chemical modification" as used herein refers to its meaning as is generally accepted in the art. With reference to exemplary nucleic acid molecules of the invention, the term refers to any modification of the chemical structure of the nucleotides that differs from nucleotides of native siRNA or RNA in general. The term "chemical modification" encompasses the addition, substitution, or modification of native siRNA or RNA at the sugar, base, or internucleotide linkage, as described herein or as is otherwise known in the art. In certain embodiments, the term "chemical modification" can refer to certain forms of RNA that are naturally occurring in certain biological systems, for example 2'-O-methyl modifications or inosine modifications.
[0073] The term "complementarity" or "complementary" as used herein refers to its meaning as is generally accepted in the art. With reference to exemplary nucleic acid molecules of the invention, the terms generally refer to the formation or existence of hydrogen bond(s) between one nucleic acid sequence and another nucleic acid sequence by either traditional Watson-Crick or other non-traditional types of bonding as described herein. In reference to the nucleic molecules of the present invention, the binding free energy for a nucleic acid molecule with its complementary sequence is sufficient to allow the relevant function of the nucleic acid to proceed, e.g., RNAi activity. Determination of binding free energies for nucleic acid molecules is well known in the art (see, e.g., Turner et al., 1987, CSH Symp. Quant. Biol. LII pp.123-133; Frier et al., 1986, Proc. Nat. Acad. Sci. USA 83:9373-9377; Turner et al., 1987, J. Am. Chem. Soc. 109:3783-3785). Perfect complementary means that all the contiguous residues of a nucleic acid sequence will hydrogen bond with the same number of contiguous residues in a second nucleic acid sequence. Partial complementarity can include various mismatches or non-based paired nucleotides (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more mismatches, non-nucleotide linkers, or non-based paired nucleotides) within the nucleic acid molecule, which can result in bulges, loops, or overhangs that result between the sense strand or sense region and the antisense strand or antisense region of the nucleic acid molecule or between the antisense strand or antisense region of the nucleic acid molecule and a corresponding target nucleic acid molecule. Such partial complementarity can be represented by a % complementarity that is determined by the number of non-base paired nucleotides, i.e., about 50%, 60%, 70%, 80%, 90% etc. depending on the total number of nucleotides involved. Such partial complementarity is permitted to the extent that the nucleic acid molecule (e.g., siNA) maintains its function, for example the ability to mediate sequence specific RNAi.
[0074] The terms "composition" or "formulation" as used herein refer to their generally accepted meaning in the art. These terms generally refer to a composition or formulation, such as in a pharmaceutically acceptable carrier or diluent, in a form suitable for administration, e.g., systemic or local administration, into a cell or subject, including, for example, a human. Suitable forms, in part, depend upon the use or the route of entry, for example oral, transdermal, inhalation, or by injection. Such forms should not prevent the composition or formulation from reaching a target cell (i.e., a cell to which the negatively charged nucleic acid is desirable for delivery). For example, compositions injected into the blood stream should be soluble. Other factors are known in the art, and include considerations such as toxicity and forms that prevent the composition or formulation from exerting its effect. As used herein, pharmaceutical formulations include formulations for human and veterinary use. Non-limiting examples of agents suitable for formulation with the nucleic acid molecules of the instant invention include: Lipid Nanoparticles (see for example Semple et al., 2010, Nat Biotechnol., Feb;28(2):172-6.); P glycoprotein inhibitors (such as Pluronic P85); biodegradable polymers, such as poly (DL lactide-coglycolide) microspheres for sustained release delivery (Emerich, DF et al, 1999, Cell Transplant, 8, 47-58); and loaded nanoparticles, such as those made of polybutylcyanoacrylate. Other non-limiting examples of delivery strategies for the nucleic acid molecules of the instant invention include material described in Boado et al., 1998, J. Pharm. Sci., 87, 1308-1315; Tyler et al., 1999, FEBS Lett., 421, 280-284; Pardridge et al., 1995, PNAS USA., 92, 5592-5596; Boado, 1995, Adv. Drug Delivery Rev., 15, 73-107; Aldrian-Herrada et al., 1998, Nucleic Acids Res., 26, 4910-4916; and Tyler et al., 1999, PNAS USA., 96, 7053-7058. A "pharmaceutically acceptable composition" or "pharmaceutically acceptable formulation" can refer to a composition or formulation that allows for the effective distribution of the nucleic acid molecules of the instant invention to the physical location most suitable for their desired activity.
[0075] The phrase "cytotoxic/cytostatic agents" refer to compounds that cause cell death or inhibit cell proliferation primarily by interfering directly with the cell's functioning or inhibit or interfere with cell mytosis, including alkylating agents, tumor necrosis factors, intercalators, hypoxia activatable compounds, microtubule inhibitors/microtubule-stabilizing agents, inhibitors of mitotic kinesins, inhibitors of histone deacetylase, inhibitors of kinases involved in mitotic progression, antimetabolites; biological response modifiers; hormonal/anti-hormonal therapeutic agents, hematopoietic growth factors, monoclonal antibody targeted therapeutic agents, topoisomerase inhibitors, proteasome inhibitors and ubiquitin ligase inhibitors.
[0076] The phrase "estrogen receptor modulators" refers to compounds that interfere with or inhibit the binding of estrogen to the receptor, regardless of mechanism.
[0077] The term "gene" or "target gene" as used herein refers to their meaning as is generally accepted in the art. The terms generally refer a nucleic acid (e.g., DNA or RNA) sequence that comprises partial length or entire length coding sequences necessary for the production of a polypeptide. The target gene can also include the UTR or non-coding region of the nucleic acid sequence. A gene or target gene can also encode a functional RNA (fRNA) or non-coding RNA (ncRNA), such as small temporal RNA (stRNA), micro RNA (miRNA), small nuclear RNA (snRNA), short interfering RNA (siRNA), small nucleolar RNA (snRNA), ribosomal RNA (rRNA), transfer RNA (tRNA) and precursor RNAs thereof. Such non-coding RNAs can serve as target nucleic acid molecules for siNA mediated RNA interference in modulating the activity of fRNA or ncRNA involved in functional or regulatory cellular processes. Aberrant fRNA or ncRNA activity leading to disease can therefore be modulated by siNA molecules of the invention. siNA molecules targeting fRNA and ncRNA can also be used to manipulate or alter the genotype or phenotype of a subject, organism or cell, by intervening in cellular processes such as genetic imprinting, transcription, translation, or nucleic acid processing (e.g., transamination, methylation etc.). The target gene can be a gene derived from a cell, an endogenous gene, a transgene, or exogenous genes such as genes of a pathogen, for example a virus, which is present in the cell after infection thereof. The cell containing the target gene can be derived from or contained in any organism, for example a plant, animal, protozoan, virus, bacterium, or fungus. Non-limiting examples of plants include monocots, dicots, or gymnosperms. Non-limiting examples of animals include vertebrates or invertebrates. Non limiting examples of fungi include molds or yeasts. For a review, see for example Snyder and Gerstein, 2003, Science, 300, 258-260.
[0078] The phrase "HMG-CoA reductase inhibitors" refers to inhibitors of 3-hydroxy-3 methylglutaryl-CoA reductase. The term HMG-CoA reductase inhibitor as used herein includes all pharmaceutically acceptable lactone and open-acid forms (i.e., where the lactone ring is opened to form the free acid) as well as salt and ester forms of compounds that have HMG-CoA reductase inhibitory activity, and therefore the use of such salts, esters, open-acid and lactone forms is included within the scope of this invention.
[0079] The term "HBV" refers to Hepatitis B Virus, which is gene that encodes HBV proteins, HBV peptides, HBV polypeptides, HBV regulatory polynucleotides (e.g., HBV miRNAs and siNAs), mutant HBV genes, and splice variants of a HBV genes, as well as other genes involved in HBV pathways of gene expression and/or activity. Thus, each of the embodiments described herein with reference to the term "HBV" are applicable to all of the protein, peptide, polypeptide, and/or polynucleotide molecules covered by the term "HBV", as that term is defined herein. Comprehensively, such gene targets are also referred to herein generally as "target" sequences (including the target sequences listed in Table la).
[0080] The phrase "highly conserved sequence region" refers to a nucleotide sequence of one or more regions in a target gene that does not vary significantly from one generation to the other or from one biological system to the other.
[0081] The phrase "homologous sequence" as used herein refers to its meaning as is generally accepted in the art. The term generally refers a nucleotide sequence that is shared by one or more polynucleotide sequences, such as genes, gene transcripts and/or non-coding polynucleotides. For example, a homologous sequence can be a nucleotide sequence that is shared by two or more genes encoding related but different proteins, such as different members of a gene family, different protein epitopes, different protein isoforms or completely divergent genes. A homologous sequence can be a nucleotide sequence that is shared by two or more non coding polynucleotides, such as noncoding DNA or RNA, regulatory sequences, introns, and sites of transcriptional control or regulation. Homologous sequences can also include sequence regions shared by more than one polynucleotide sequence. Homology does not need to be perfect identity (100%), as partially homologous sequences are also contemplated by and within the scope of the instant invention (e.g., at least 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80% etc.). Percent homology is the number of matching nucleotides between two sequences divided by the total length being compared, multiplied by 100.
[0082] The phrase "improved RNAi activity" refers to an increase in RNAi activity measured in vitro and/or in vivo, where the RNAi activity is a reflection of both the ability of the siNA to mediate RNAi and the stability of the siNAs of the invention. In this invention, the product of these activities can be increased in vitro and/or in vivo compared to an all RNA siNA or an siNA containing a plurality of ribonucleotides. In some cases, the activity or stability of the siNA molecule can be decreased (i.e., less than ten-fold), but the overall activity of the siNA molecule is enhanced in vitro and/or in vivo.
[0083] The terms "inhibit," "down-regulate," or "reduce" as used herein refers to its meaning as is generally accepted in the art. With reference to exemplary nucleic acid molecules of the invention, he term generally refers the reduction in the expression of the gene, or level of RNA molecules or equivalent RNA molecules encoding one or more proteins or protein subunits, or activity of one or more proteins or protein subunits, below that observed in the absence of the nucleic acid molecules (e.g., siNA) of the invention. Down-regulation can also be associated with post-transcriptional silencing, such as, RNAi mediated cleavage or by alteration in DNA methylation patterns or DNA chromatin structure. Inhibition, down-regulation or reduction with an siNA molecule can be in reference to an inactive molecule, an attenuated molecule, an siNA molecule with a scrambled sequence, or an siNA molecule with mismatches or alternatively, it can be in reference to the system in the absence of the nucleic acid.
[0084] The phrase "inhibitors of cell proliferation and survival signaling pathway" refers to pharmaceutical agents that inhibit cell surface receptors and signal transduction cascades downstream of those surface receptors.
[0085] The term "integrin blockers" refers to compounds which selectively antagonize, inhibit or counteract binding of a physiological ligand to the a@3 integrin, to compounds which
selectively antagonize, inhibit or counteract binding of a physiological ligand to the am35 integrin, to compounds which antagonize, inhibit or counteract binding of a physiological ligand to both the am@3 integrin and the amPs integrin, and to compounds which antagonize, inhibit or counteract the activity of the particular integrin(s) expressed on capillary endothelial cells. The term also refers to antagonists of the aU4.6 am.8 ip.1 a 2 0.1 a5p.1 ac6pi and c6P4 integrins. The term
also refers to antagonists of any combination Of amP3, ams, a@.6 am@.8 aip.1 a2 0.1 5P.1 acpi and
a6P4 integrins.
[0086] The terms "intermittent" or "intermittently" as used herein refers to its meaning as is generally accepted in the art. The term generally refers to periodic stopping and starting at either regular or irregular intervals.
[0087] The terms "internucleoside linkage" or "internucleoside linker" or "intemucleotide linkage" or "intemucleotide linker" are used herein interchangeably and refer to any linker or linkage between two nucleoside units, as is known in the art, including, for example, but not limitation, phosphate, analogs of phosphate, phosphonate, guanidium, hydroxylamine, hydroxylhydrazinyl, amide, carbamate, alkyl, and substituted alkyl linkages. The internucleoside linkages constitute the backbone of a nucleic acid molecule.
[0088] The terms "mammalian" or "mammal" as used herein refers to its meaning as is generally accepted in the art. The term generally refers to any warm blooded vertebrate species, such as a human, mouse, rat, dog, cat, hamster, guinea pig, rabbit, livestock, and the like.
[0089] The phrase "metered dose inhaler" or MDI refers to a unit comprising a can, a secured cap covering the can and a formulation metering valve situated in the cap. MDI systems includes a suitable channeling device. Suitable channeling devices comprise for example, a valve actuator and a cylindrical or cone-like passage through which medicament can be delivered from the filled canister via the metering valve to the nose or mouth of a patient such as a mouthpiece actuator.
[0090] The term "microRNA" or "miRNA" as used herein refers to its meaning as is generally accepted in the art. The term generally refers a small double-stranded RNA that regulates the expression of target messenger RNAs either bymRNA cleavage, translational repression/inhibition or heterochromatic silencing (see for example Ambros, 2004, Nature, 431, 350-355; Bartel, 2004, Cell, 116, 281-297; Cullen, 2004, Virus Research., 102, 3-9; He et al., 2004, Nat. Rev. Genet., 5, 522-531; Ying et al., 2004, Gene, 342, 25-28; and Sethupathy et al., 2006, RNA, 12:192-197).
[0091] The term "modulate" as used herein refers to its meaning as is generally accepted in the art. With reference to exemplary nucleic acid molecules of the invention, the term refers to when the expression of a gene, or level of one or more RNA molecules (coding or non-coding), or activity of one or more RNA molecules or proteins or protein subunits, is up-regulated or down-regulated, such that expression, level, or activity is greater than or less than that observed in the absence of the molecule that effects modulation. For example, the term "modulate" in some embodiments can refer to inhibition and in other embodiments can refer to potentiation or up-regulation, e.g., of gene expression.
[0092] The phrase "modified nucleotide" as used herein refers to its meaning as is generally accepted in the art. The term generally refers a nucleotide, which contains a modification in the chemical structure of the base, sugar and/or phosphate of the unmodified (or natural) nucleotide as is generally known in the art. Non-limiting examples of modified nucleotides are described herein and in U.S. Application No. 12/064,014.
[0093] The phrase "NSAIDs that are selective COX-2 inhibitors" for purposes herein, refers to NSAIDs, which possess a specificity for inhibiting COX-2 over COX-1 of at least 100 fold as measured by the ratio of IC 5 0 for COX-2 over IC 5 0 for COX-1 evaluated by cell or microsomal
assays.
[0094] The phrase "non-base paired" refers to nucleotides that are not base paired between the sense strand or sense region and the antisense strand or antisense region of an double- stranded siNA molecule; and can include for example, but not limitation, mismatches, overhangs, single stranded loops, etc.
[0095] The term "non-nucleotide" refers to any group or compound which can be incorporated into a nucleic acid chain in the place of one or more nucleotide units, such as for example but not limitation abasic moieties or alkyl chains. The group or compound is "abasic" in that it does not contain a commonly recognized nucleotide base, such as adenosine, guanine, cytosine, uracil or thymine and therefore lacks a nucleobase at the '-position.
[0096] The term "nucleotide" is used as is generally recognized in the art. Nucleotides generally comprise a nucleobase, a sugar, and an internucleoside linkage, e.g., a phosphate. The base can be a natural bases (standard), modified bases, or a base analog, as are well known in the art. Such bases are generally located at the ' position of a nucleotide sugar moiety. Additionally, the nucleotides can be unmodified or modified at the sugar, internucleoside linkage, and/or base moiety, (also referred to interchangeably as nucleotide analogs, modified nucleotides, non-natural nucleotides, non-standard nucleotides and others; see, for example, U.S. Application No. 12/064,014.
[0097] The term "overhang" as used herein refers to its meaning as is generally accepted in the art. With reference to exemplary double stranded nucleic acid molecules, the term generally refers to the terminal portion of a nucleotide sequence that is not base paired between the two strands of a double-stranded nucleic acid molecule (see for example, Figure 4). Overhangs, when present, are typically at the 3'-end of one or both strands in a siNA duplex.
[0098] The term "parenteral" as used herein refers to its meaning as is generally accepted in the art. The term generally refers methods or techniques of administering a molecule, drug, agent, or compound in a manner other than through the digestive tract, and includes epicutaneous, subcutaneous, intravascular (e.g., intravenous), intramuscular, or intrathecal injection or infusion techniques and the like.
[0099] The phrase "pathway target" refers to any target involved in pathways of gene expression or activity. For example, any given target can have related pathway targets that can include upstream, downstream, or modifier genes in a biologic pathway. These pathway target genes can provide additive or synergistic effects in the treatment of diseases, conditions, and traits herein.
[0100] The term "phosphorothioate" refers to an internucleotide phosphate linkage comprising one or more sulfur atoms in place of an oxygen atom. Hence, the term phosphorothioate refers to both phosphorothioate and phosphorodithioate internucleotide linkages.
[0101] "Prenyl-protein transferase inhibitor" refers to a compound that inhibits any one or any combination of the prenyl-protein transferase enzymes, including farnesyl-protein transferase (FPTase), geranylgeranyl-protein transferase type I (GGPTase-I), and geranylgeranyl-protein transferase type-II (GGPTase-II, also called Rab GGPTase).
[0102] The phrase "retinoid receptor modulators" refers to compounds that interfere or inhibit the binding of retinoids to the receptor, regardless of mechanism.
[0103] The term "ribonucleotide" as used herein refers to its meaning as is generally accepted in the art. The term generally refers to a nucleotide with a hydroxyl group at the 2' position of a p-D-ribofuranose moiety.
[0104] The term "RNA" as used herein refers to its generally accepted meaning in the art. Generally, the term RNA refers to a molecule comprising at least one ribofuranoside moiety. The term can include double-stranded RNA, single-stranded RNA, isolated RNA such as partially purified RNA, essentially pure RNA, synthetic RNA, recombinantly produced RNA, as well as altered RNA that differs from naturally occurring RNA by the addition, deletion, substitution and/or alteration of one or more nucleotides. Such alterations can include addition of non-nucleotide material, such as to the end(s) of the siNA or internally, for example at one or more nucleotides of the RNA. Nucleotides in the RNA molecules of the instant invention can also comprise non-standard nucleotides, such as non-naturally occurring nucleotides or chemically synthesized nucleotides or deoxynucleotides. These altered RNAs can be referred to as analogs or analogs of naturally-occurring RNA.
[0105] The phrase "RNA interference" or term "RNAi" refer to the biological process of inhibiting or down regulating gene expression in a cell, as is generally known in the art, and which is mediated by short interfering nucleic acid molecules, see for example Zamore and Haley, 2005, Science, 309, 1519-1524; Vaughn and Martienssen, 2005, Science, 309, 1525-1526; Zamore et al., 2000, Cell, 101, 25-33; Bass, 2001, Nature, 411, 428-429; Elbashir et al., 2001, Nature, 411, 494-498; and Kreutzer et al., International PCT Publication No. WO 00/44895; Zernicka-Goetz et al., International PCT Publication No. WO 01/36646; Fire, International PCT Publication No. WO 99/32619; Plaetinck et al., International PCT Publication No. WO /01846; Mello and Fire, International PCT Publication No. WO 01/29058; Deschamps Depaillette, International PCT Publication No. WO 99/07409; and Li et al., International PCT Publication No. WO 00/44914; Allshire, 2002, Science, 297, 1818-1819; Volpe et al., 2002, Science, 297, 1833-1837; Jenuwein, 2002, Science, 297, 2215-2218; and Hall et al., 2002, Science, 297, 2232-2237; Hutvagner and Zamore, 2002, Science, 297, 2056-60; McManus et al., 2002, RNA, 8, 842-850; Reinhart et al., 2002, Gene & Dev., 16, 1616-1626; and Reinhart
& Bartel, 2002, Science, 297, 1831). Additionally, the term RNAi is meant to be equivalent to other terms used to describe sequence specific RNA interference, such as post transcriptional gene silencing, translational inhibition, transcriptional inhibition, or epigenetics. For example, siNA molecules of the invention can be used to epigenetically silence genes at either the post transcriptional level or the pre-transcriptional level. In a non-limiting example, epigenetic modulation of gene expression by siNA molecules of the invention can result from siNA mediated modification of chromatin structure or methylation patterns to alter gene expression (see, for example, Verdel et al., 2004, Science, 303, 672-676; Pal-Bhadra et al., 2004, Science, 303, 669-672; Allshire, 2002, Science, 297, 1818-1819; Volpe et al., 2002, Science, 297, 1833 1837; Jenuwein, 2002, Science, 297, 2215-2218; and Hall et al., 2002, Science, 297, 2232-2237). In another non-limiting example, modulation of gene expression by siNA molecules of the invention can result from siNA mediated cleavage of RNA (either coding or non-coding RNA) via RISC, or via translational inhibition, as is known in the art or modulation can result from transcriptional inhibition (see for example Janowski et al., 2005, Nature Chemical Biology, 1, 216-222).
[0106] The phrase "RNAi inhibitor" refers to any molecule that can down regulate, reduce or inhibit RNA interference function or activity in a cell or organism. An RNAi inhibitor can down regulate, reduce or inhibit RNAi (e.g., RNAi mediated cleavage of a target polynucleotide, translational inhibition, or transcriptional silencing) by interaction with or interfering with the function of any component of the RNAi pathway, including protein components such as RISC, or nucleic acid components such as miRNAs or siRNAs. A RNAi inhibitor can be an siNA molecule, an antisense molecule, an aptamer, or a small molecule that interacts with or interferes with the function of RISC, a miRNA, or an siRNA or any other component of the RNAi pathway in a cell or organism. By inhibiting RNAi (e.g., RNAi mediated cleavage of a target polynucleotide, translational inhibition, or transcriptional silencing), a RNAi inhibitor of the invention can be used to modulate (e.g., up-regulate or down regulate) the expression of a target gene.
[0107] The phrase "sense region" as used herein refers to its meaning as is generally accepted in the art. With reference to exemplary nucleic acid molecules of the invention, the term refers to a nucleotide sequence of an siNA molecule having complementarity to an antisense region of the siNA molecule. In addition, the sense region of an siNA molecule can comprise a nucleic acid sequence having homology or sequence identity with a target nucleic acid sequence. In one embodiment, the sense region of the siNA molecule is also referred to as the sense strand or passenger strand.
[0108] The phrases "short interfering nucleic acid", "siNA", "short interfering RNA", "siRNA", "short interfering nucleic acid molecule", "short interfering oligonucleotide molecule", or "chemically modified short interfering nucleic acid molecule" refer to any nucleic acid molecule capable of inhibiting or down regulating gene expression or viral replication by mediating RNA interference ("RNAi") or gene silencing in a sequence-specific manner. These terms can refer to both individual nucleic acid molecules, a plurality of such nucleic acid molecules, or pools of such nucleic acid molecules. The siNA can be a double-stranded nucleic acid molecule comprising self-complementary sense and antisense strands, wherein the antisense strand comprises a nucleotide sequence that is complementary to a nucleotide sequence in a target nucleic acid molecule or a portion thereof and the sense strand comprises a nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof. The siNA can be a polynucleotide with a duplex, asymmetric duplex, hairpin or asymmetric hairpin secondary structure, having self-complementary sense and antisense regions, wherein the antisense region comprises a nucleotide sequence that is complementary to a nucleotide sequence in a separate target nucleic acid molecule or a portion thereof and the sense region comprises a nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof. The siNA can be a circular single-stranded polynucleotide having two or more loop structures and a stem comprising self-complementary sense and antisense regions, wherein the antisense region comprises nucleotide sequence that is complementary to a nucleotide sequence in a target nucleic acid molecule or a portion thereof and the sense region comprises a nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof, and wherein the circular polynucleotide can be processed either in vivo or in vitro to generate an active siNA molecule capable of mediating RNAi. The siNA can also comprise a single-stranded polynucleotide having a nucleotide sequence complementary to nucleotide sequence in a target nucleic acid molecule or a portion thereof (for example, where such siNA molecule does not require the presence within the siNA molecule of a nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof), wherein the single-stranded polynucleotide can further comprise a terminal phosphate group, such as a 5'-phosphate (see for example, Martinez et al., 2002, Cell, 110, 563-574 and Schwarz et al., 2002, Molecular Cell, 10, 537-568), or 5',3' diphosphate.
[0109] The term "subject" as used herein refers to its meaning as is generally accepted in the art. The term generally refers an organism to which the nucleic acid molecules of the invention can be administered. A subject can be a mammal or mammalian cells, including a human or human cells. The term also refers to an organism, which is a donor or recipient of explanted cells or the cells themselves.
[0110] The phrase "systemic administration" as used herein refers to its meaning as is generally accepted in the art. The term generally refers in vivo systemic absorption or accumulation of drugs in the blood stream followed by distribution throughout the entire body.
[0111] The term "target" as it refers to HBV refers to any HBV target protein, peptide, or polypeptide, such as encoded by Genbank Accession Nos. shown in Table 7. The term also refers to nucleic acid sequences or target polynucleotide sequence encoding any target protein, peptide, or polypeptide, such as proteins, peptides, or polypeptides encoded by sequences having Genbank Accession Nos. shown in Table 7. The target of interest can include target polynucleotide sequences, such as target DNA or target RNA. The term "target" is also meant to include other sequences, such as differing isoforms, mutant target genes, splice variants of target polynucleotides, target polymorphisms, and non-coding (e.g., ncRNA, miRNA, stRNA, sRNA) or other regulatory polynucleotide sequences as described herein.
[0112] The phrase "target site" as used herein refers to its meaning as is generally accepted in the art. The term generally refers to a sequence within a target nucleic acid molecule, (e.g., RNA) that is "targeted", e.g., for cleavage mediated by an siNA construct, which contains sequences within its antisense region that are complementary to the target sequence.
[0113] The phrase "therapeutically effective amount" as used herein refers to its meaning as is generally accepted in the art. The term generally refers to the amount of the compound or composition that will elicit the biological or medical response of a cell, tissue, system, animal or human that is be sought by the researcher, veterinarian, medical doctor or other clinician. For example, if a given clinical treatment is considered effective when there is at least a 25% reduction in a measurable parameter associated with a disease or disorder, a therapeutically effective amount of a drug for the treatment of that disease or disorder is that amount necessary to effect at least a 25% reduction in that parameter.
[0114] The phrase "universal base" as used herein refers to its meaning as is generally accepted in the art. The term universal base generally refers to nucleotide base analogs that form base pairs with each of the natural DNA/RNA bases with little or no discrimination between them. Non-limiting examples of universal bases include C-phenyl, C-naphthyl and other aromatic derivatives, inosine, azole carboxamides, and nitroazole derivatives such as 3 nitropyrrole, 4-nitroindole, 5-nitroindole, and 6-nitroindole as known in the art (see for example, Loakes, 2001, Nucleic Acids Research, 29, 2437-2447).
[0115] The term "up-regulate" as used herein refers to its meaning as is generally accepted in the art. With reference to exemplary nucleic acid molecules of the invention, the term refers to an increase in the expression of a gene, or level of RNA molecules or equivalent RNA molecules encoding one or more proteins or protein subunits, or activity of one or more RNAs, proteins or protein subunits, above that observed in the absence of the nucleic acid molecules (e.g., siNA) of the invention. In certain instances, up-regulation or promotion of gene expression with an siNA molecule is above that level observed in the presence of an inactive or attenuated molecule. In other instances, up-regulation or promotion of gene expression with siNA molecules is above that level observed in the presence of, for example, an siNA molecule with scrambled sequence or with mismatches. In still other instances, up-regulation or promotion of gene expression with a nucleic acid molecule of the instant invention is greater in the presence of the nucleic acid molecule than in its absence. In some instances, up-regulation or promotion of gene expression is associated with inhibition of RNA mediated gene silencing, such as RNAi mediated cleavage or silencing of a coding or non-coding RNA target that down regulates, inhibits, or silences the expression of the gene of interest to be up-regulated. The down regulation of gene expression can, for example, be induced by a coding RNA or its encoded protein, such as through negative feedback or antagonistic effects. The down regulation of gene expression can, for example, be induced by a non-coding RNA having regulatory control over a gene of interest, for example by silencing expression of the gene via translational inhibition, chromatin structure, methylation, RISC mediated RNA cleavage, or translational inhibition. As such, inhibition or down regulation of targets that down regulate, suppress, or silence a gene of interest can be used to up regulate expression of the gene of interest toward therapeutic use.
[0116] The term "vector" as used herein refers to its meaning as is generally accepted in the art. The term vector generally refers to any nucleic acid- and/or viral-based expression system or technique used to deliver one or more nucleic acid molecules.
B. siNA Molecules of the Invention
[0117] The present invention provides compositions and methods comprising siNAs targeted to HBV that can be used to treat diseases, e.g., liver disease or malignancies and/or cancers associated with HBV expression or other conditions that result from hepatitis B infection. In particular aspects and embodiments of the invention, the nucleic acid molecules of the invention comprise at least a 15 nucleotide sequence of the sequences shown in Table la and Table 1b. The siNAs can be provided in several forms. For example, the siNA can be isolated as one or more siNA compounds, or it may be in the form of a transcriptional cassette in a DNA plasmid. The siNA may also be chemically synthesized and can include modifications as shown, for example, but not limitation, in Table le and Table 8. Thus, in various embodiments, at least one strand or region of the nucleic acids of the invention comprises at least a 15 nucleotide sequence selected from the group of sequences consisting of SEQ ID NOS:1-502. The siNAs can be administered alone or co-administered with other siNA molecules or with conventional agents that treat a HBV related disease or condition.
[0118] The siNA molecules of the invention can be used to mediate gene silencing, specifically HBV, via interaction with RNA transcripts or alternately by interaction with particular gene sequences, wherein such interaction results in modulation of gene silencing either at the transcriptional level or post-transcriptional level such as, for example, but not limited to, RNAi or through cellular processes that modulate the chromatin structure or methylation patterns of the target and prevent transcription of the target gene, with the nucleotide sequence of the target thereby mediating silencing. More specifically, the target is any of HBV RNA, DNA, or mRNA,
[0119] In one aspect, the invention provides short interfering nucleic acid (siNA) molecules for inhibiting the expression of the HBV gene in a cell or mammal. The siNA can be single stranded or double-stranded. When double-stranded, the siNA comprising a sense and an antisense stand. The antisense strand is complementary to at least a part of an mRNA formed in the expression of the HBV gene. The sense strand comprises a region that is complementary to the antisense strand. In specific embodiments, the antisense strand comprises at least a 15 nucleotide sequence of an antisense sequence listed in Table 1b. Generally, the double-stranded siNA comprises at least a 15 nucleotide sequence of the sense strand in Table lb and at least a nucleotide sequence of the antisense strand in Table 1b. One or more of the nucleotides of the siNAs of the invention are optionally modified. In further embodiments having modifications, some siNAs of the invention comprises at least one nucleotide sequence selected from the groups of sequences provide in Table 1c. In other embodiments, the siNA comprises at least two sequences selected from the group of sequences provided in Table 1c, wherein one of the at least two sequences is complementary to another of the at least two sequences and one of the at least two sequences is complementary to a sequence of a mRNA generated in the expression of the HBV gene. Examples of certain modified siNAs of the invention are in Table 1c.
[0120] The double stranded RNA molecules of the invention can comprise two distinct and separate strands that can be symmetric or asymmetric and are complementary, i.e., two single stranded RNA molecules, or can comprise one single-stranded molecule in which two complementary portions, e.g., a sense region and an antisense region, are base-paired, and are covalently linked by one or more single-stranded "hairpin" areas (i.e. loops) resulting in, for example, a single-stranded short-hairpin polynucleotide or a circular single-stranded polynucleotide.
[0121] The linker can be polynucleotide linker or a non-nucleotide linker. In some embodiments, the linker is a non-nucleotide linker. In some embodiments, a hairpin or circular siNA molecule of the invention contains one or more loop motifs, wherein at least one of the loop portions of the siNA molecule is biodegradable. For example, a single-stranded hairpin siNA molecule of the invention is designed such that degradation of the loop portion of the siNA molecule in vivo can generate a double-stranded siNA molecule with 3'-terminal overhangs, such as 3'-terminal nucleotide overhangs comprising 1, 2, 3 or 4 nucleotides. Or alternatively, a circular siNA molecule of the invention is designed such that degradation of the loop portions of the siNA molecule in vivo can generate a double-stranded siNA molecule with 3'-terminal overhangs, such as 3'-terminal nucleotide overhangs comprising about 2 nucleotides.
[0122] In symmetric siNA molecules of the invention, each strand, the sense (passenger) strand and antisense (guide) strand, are independently about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides in length. Generally, each strand of the symmetric siNA molecules of the invention are about 19-24 (e.g., about 19, 20, 21, 22, 23 or 24) nucleotides in length.
[0123] In asymmetric siNA molecules, the antisense region or strand of the molecule is about to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides in length, wherein the sense region is about 3 to about 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) nucleotides in length. Generally, each strand of the asymmetric siNA molecules of the invention is about 19-24 (e.g., about 19, 20, 21, 22, 23 or 24) nucleotides in length.
[0124] In yet other embodiments, siNA molecules of the invention comprise single stranded hairpin siNA molecules, wherein the siNA molecules are about 25 to about 70 (e.g., about 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 40, 45, 50, 55, 60, 65, or 70) nucleotides in length.
[0125] In still other embodiments, siNA molecules of the invention comprise single-stranded circular siNA molecules, wherein the siNA molecules are about 38 to about 70 (e.g., about 38, , 45, 50, 55, 60, 65, or 70) nucleotides in length.
[0126] In still other embodiments, siNA molecules of the invention comprise single-stranded non-circular siNA molecules, wherein the siNA molecules are independently about 15 to about (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides in length.
[0127] In various symmetric embodiments, the siNA duplexes of the invention independently comprise about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, , 26, 27, 28, 29, or 30) base pairs. Generally, the duplex structure of siNAs of the invention is between 15 and 30, more generally between 18 and 25, yet more generally between 19 and 24, and most generally between 19 and 21 base pairs in length.
[0128] In yet other embodiments, where the duplex siNA molecules of the invention are asymmetric, the siNA molecules comprise about 3 to 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) base pairs. Generally, the duplex structure of siNAs of the invention is between 15 and 25, more generally between 18 and 25, yet more generally between 19 and 24, and most generally between 19 and 21 base pairs in length.
[0129] In still other embodiments, where the siNA molecules of the invention are hairpin or circular structures, the siNA molecules comprise about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) base pairs.
[0130] The sense strand and antisense strand, or the sense region and antisense region, of the siNA molecules of the invention can be complementary. Also, the antisense strand or antisense region can be complementary to a nucleotide sequence or a portion thereof of the HBV target RNA. The sense strand or sense region of the siNA can comprise a nucleotide sequence of a HBV gene or a portion thereof. In certain embodiments, the sense region or sense strand of an siNA molecule of the invention is complementary to that portion of the antisense region or antisense strand of the siNA molecule that is complementary to a HBV target polynucleotide sequence, such as for example, but not limited to, those sequences represented by GENBANK Accession Nos. shown in Table 7.
[0131] In some embodiments, siNA molecules of the invention have perfect complementarity between the sense strand or sense region and the antisense strand or antisense region of the siNA molecule. In other or the same embodiments, the antisense strand of the siNA molecules of the invention are perfectly complementary to a corresponding target nucleic acid molecule.
[0132] In yet other embodiments, siNA molecules of the invention have partial complementarity (i.e., less than 100% complementarity) between the sense strand or sense region and the antisense strand or antisense region of the siNA molecule or between the antisense strand or antisense region of the siNA molecule and a corresponding target nucleic acid molecule. Thus, in some embodiments, the double-stranded nucleic acid molecules of the invention, have between about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides in one strand that are complementary to the nucleotides of the other strand. In other embodiments, the molecules have between about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides in the sense region that are complementary to the nucleotides of the antisense region. of the double-stranded nucleic acid molecule. In certain embodiments, the double-stranded nucleic acid molecules of the invention have between about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides in the antisense strand that are complementary to a nucleotide sequence of its corresponding target nucleic acid molecule.
[0133] In other embodiments, the siNA molecule can contain one or more nucleotide deletions, substitutions, mismatches and/or additions; provided, however, that the siNA molecule maintains its activity, for example, to mediate RNAi. In a non-limiting example, the deletion, substitution, mismatch and/or addition can result in a loop or bulge, or alternately a wobble or other alternative (non Watson-Crick) base pair. Thus, in some embodiments, for example, the double-stranded nucleic acid molecules of the invention, have 1or more (e.g., 1, 2, 3, 4, 5, or 6) nucleotides, in one strand or region that are mismatches or non-base-paired with the other strand or region.. In other embodiments, the double-stranded nucleic acid molecules of the invention, have 1 or more (e.g., 1, 2, 3, 4, 5, or 6) nucleotides in each strand or region that are mismatches or non-base-paired with the other strand or region. In a preferred embodiment, the siNA of the invention contains no more than 3 mismatches. If the antisense strand of the siNA contains mismatches to a target sequence, it is preferable that the area of mismatch not be located in the center of the region of complementarity.
[0134] In other embodiments, the siNA molecule can contain one or more nucleotide deletions, substitutions, mismatches and/or additions to a sequence in Table lb provided, however, that the siNA molecule maintains its activity, for example, to mediate RNAi. In a non limiting example, the deletion, substitution, mismatch and/or addition can result in a loop or bulge, or alternately a wobble or other alternative (non Watson-Crick) base pair.
[0135] The invention also comprises double-stranded nucleic acid (siNA) molecules as otherwise described hereinabove in which the first strand and second strand are complementary to each other and wherein at least one strand is hybridisable to the polynucleotide sequence of a sequence in Table lb under conditions of high stringency, and wherein any of the nucleotides is unmodified or chemically modified.
[0136] Hybridization techniques are well known to the skilled artisan (see for instance, Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989)). Preferred stringent hybridization conditions include overnight incubation at 42°C in a solution comprising: 50% formamide, xSSC (150mM NaCl, 15mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5x Denhardt's solution, 10% dextran sulfate, and 20 microgram/ml denatured, sheared salmon sperm DNA; followed by washing the filters in 0.1x SSC at about 65°C.
[0137] In one specific embodiment, the first strand has about 15, 16, 17, 18, 19, 20 or 21 nucleotides that are complementary to the nucleotides of the other strand and at least one strand is hybridisable to a polynucleotide sequence in Table 1b. In a more preferred embodiment, the first strand has about 15, 16, 17, 18, 19, 20 or 21 nucleotides that are complementary to the nucleotides of the other strand and at least one strand is hybridisable to SEQ ID NO: 1, SEQ ID
NO: 1210, SEQ ID NO: 2, SEQ ID NO: 1211, SEQ ID NO: 3, SEQ ID NO: 1212, SEQ ID NO: 4, or SEQ ID NO:1213; under conditions of high stringency, and wherein any of the nucleotides is unmodified or chemically modified.
[0138] In certain embodiments, the siNA molecules of the invention comprise overhangs of about 1 to about 4 (e.g., about 1, 2, 3 or 4) nucleotides. The nucleotides in the overhangs can be the same or different nucleotides. In some embodiments, the overhangs occur at the 3'-end at one or both strands of the double-stranded nucleic acid molecule. For example, a double stranded nucleic acid molecule of the invention can comprise a nucleotide or non-nucleotide overhang at the 3'-end of the antisense strand/region, the 3'-end of the sense strand/region, or both the antisense strand/region and the sense strand/region of the double-stranded nucleic acid molecule.
[0139] In some embodiments, the nucleotides comprising the overhang portion of an siNA molecule of the invention comprise sequences based on the HBV target polynucleotide sequence in which nucleotides comprising the overhang portion of the antisense strand/region of an siNA molecule of the invention can be complementary to nucleotides in the HBV target polynucleotide sequence and/or nucleotides comprising the overhang portion of the sense strand/region of an siNA molecule of the invention can comprise the nucleotides in the HBV target polynucleotide sequence. Thus, in some embodiments, the overhang comprises a two nucleotide overhang that is complementary to a portion of the HBV target polynucleotide sequence. In other embodiments, however, the overhang comprises a two nucleotide overhang that is not complementary to a portion of the HBV target polynucleotide sequence. In certain embodiments, the overhang comprises a 3'-UU overhang that is not complementary to a portion of the HBV target polynucleotide sequence. In other embodiments, the overhang comprises a UU overhang at the 3' end of the antisense strand and a TT overhang at the 3' end of the sense strand. In other embodiments, the overhang comprises nucleotides as described in the examples, Tables, and Figures herein.
[0140] In any of the embodiments of the siNA molecules described herein having 3'-terminal nucleotide overhangs, the overhangs are optionally chemically modified at one or more nucleic acid sugar, base, or backbone positions. Representative, but not limiting examples of modified nucleotides in the overhang portion of a double-stranded nucleic acid (siNA) molecule of the invention include: 2'-O-alkyl (e.g., 2'-O-methyl), 2'-deoxy, 2'-deoxy-2'-fluoro, 2'-deoxy-2' fluoroarabino (FANA), 4'-thio, 2'-O-trifluoromethyl, 2'-O-ethyl-trifluoromethoxy, 2'-0 difluoromethoxy-ethoxy, universal base, acyclic, or 5-C-methyl nucleotides. In more preferred embodiments, the overhang nucleotides are each independently, a 2'-O-alkyl nucleotide, a 2'-0 methyl nucleotide, a 2'-dexoy-2-fluoro nucleotide, or a 2'-deoxy ribonucleotide. In some instances the overhang nucleotides are linked by a one or more phosphorothioate linkages.
[0141] In yet other embodiments, siNA molecules of the invention comprise duplex nucleic acid molecules with blunt ends (i.e., without nucleotide overhangs), where both ends are blunt, or alternatively, where one of the ends is blunt. In some embodiments, the siNA molecules of the invention can comprises one blunt end, for example wherein the 5'-end of the antisense strand and the 3'-end of the sense strand do not have any overhanging nucleotides. In another example, the siNA molecule comprises one blunt end, for example wherein the 3'-end of the antisense strand and the 5'-end of the sense strand do not have any overhanging nucleotides. In other embodiments, siNA molecules of the invention comprise two blunt ends, for example wherein the 3'-end of the antisense strand and the 5'-end of the sense strand as well as the 5'-end of the antisense strand and 3'-end of the sense strand do not have any overhanging nucleotides.
[0142] In any of the embodiments or aspects of the siNA molecules of the invention, the sense strand and/or the antisense strand can further have a cap, such as described herein or as known in the art, at the 3'-end, the 5'-end, or both of the 3' and 5'-ends of the sense strand and/or antisense strand. Or as in the case of a hairpin siNA molecule, the cap can be at either one or both of the terminal nucleotides of the polynucleotide. In some embodiments, the cap is at one of both of the ends of the sense strand of a double-stranded siNA molecule. In other embodiments, the cap is at the 3'-end of antisense (guide) strand. In preferred embodiments, the caps are at the 3'-end of the sense strand and the 5'-end of the sense strand.
[0143] Representative, but non-limiting examples of such terminal caps include an inverted abasic nucleotide, an inverted deoxy abasic nucleotide, an inverted nucleotide moiety, a group shown in Figure 5, a glyceryl modification, an alkyl or cycloalkyl group, a heterocycle, or any other cap as is generally known in the art.
[0144] Any of the embodiments of the siNA molecules of the invention can have a 5' phosphate termini. In some embodiments, the siNA molecules lack terminal phosphates.
[0145] Any siNA molecule or construct of the invention can comprise one or more chemical modifications. Modifications can be used to improve in vitro or in vivo characteristics such as stability, activity, toxicity, immune response (e.g., prevent stimulation of an interferon response, an inflammatory or pro-inflammatory cytokine response, or a Toll-like Receptor (TF) response), and/or bioavailability.
[0146] Applicants describe herein chemically modified siNA molecules with improved RNAi activity and/or stability compared to corresponding unmodified siNA molecules. Various chemically modified siNA motifs disclosed herein provide the capacity to maintain RNAi activity that is substantially similar to unmodified or minimally modified active siRNA (see for example Elbashir et al., 2001, EMBO J., 20:6877-6888) while at the same time providing nuclease resistance and pharmacokinetic properties suitable for use in therapeutic applications.
[0147] In various embodiments, the siNA molecules of the invention comprise modifications wherein any (e.g., one or more or all) nucleotides present in the sense and/or antisense strand are modified nucleotides (e.g., wherein one nucleotide is modified, some nucleotides (i.e., plurality or more than one) are modified, or all nucleotides are modified nucleotides. In some embodiments, the siNA molecules of the invention are partially modified (e.g., about 1, 2, 3, 4, ,6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20,21,22,23,24,25,26,27,28,29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 55, or 59 nucleotides are modified) with chemical modifications. In some embodiments, an siNA molecule of the invention comprises at least about 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, or 60 nucleotides that are modified nucleotides. In other embodiments, the siNA molecules of the invention are completely modified (e.g., 100% modified) with chemical modifications, i.e., the siNA molecule does not contain any ribonucleotides. In some of embodiments, one or more of the nucleotides in the sense strand of the siNA molecules of the invention are modified. In the same or other embodiments, one or more of the nucleotides in the antisense strand of the siNA molecules of the invention are modified.
[0148] The chemical modification within a single siNA molecule can be the same or different. In some embodiments, at least one strand has at least one chemical modification. In other embodiments, each strand has at least one chemical modifications, which can be the same or different, such as, sugar, base, or backbone (i.e., internucleotide linkage) modifications. In other embodiments, siNA molecules of the invention contain at least 2, 3, 4, 5, or more different chemical modifications.
[0149] Non-limiting examples of chemical modifications that are suitable for use in the present invention, are disclosed in U.S. Patent Applications Nos. 10/444,853; 10/981,966; 12/064,014 and in references cited therein and include sugar, base, and phosphate, non nucleotide modifications, and/or any combination thereof.
[0150] In certain specific embodiments of the invention, at least one modified nucleotide is a 2'-deoxy-2-fluoro nucleotide, a 2'-deoxy nucleotide, a 2'-O-alkyl (e.g., 2'-O-methyl) nucleotide, or a locked nucleic acid (LNA) nucleotide as is generally recognized in the art.
[0151] In yet other embodiment of the invention, at least one nucleotide has a ribo-like, Northern or A form helix configuration (see e.g., Saenger, Principles of Nucleic Acid Structure, Springer-Verlag ed., 1984). Non-limiting examples of nucleotides having a Northern configuration include locked nucleic acid (LNA) nucleotides (e.g., 2'-O, 4'-C-methylene-(D ribofuranosyl) nucleotides); 2'-methoxyethoxy (MOE) nucleotides; 2'-methyl-thio-ethyl nucleotides, 2'-deoxy-2'-fluoro nucleotides; 2'-deoxy-2'-chloro nucleotides; 2'-azido nucleotides; 2'-O-trifluoromethyl nucleotides; 2'-O-ethyl-trifluoromethoxy nucleotides; 2'-0 difluoromethoxy-ethoxy nucleotides; 4'-thio nucleotides and 2'-O-methyl nucleotides.
[0152] In various embodiments, a majority (e.g., greater than 50%) of the pyrimidine nucleotides present in the double-stranded siNA molecule comprises a sugar modification. In some of the same and/or other embodiments, a majority (e.g., greater than 50%) of the purine nucleotides present in the double-stranded siNA molecule comprises a sugar modification.
[0153] In some embodiments, the pyrimidine nucleotides in the antisense strand are 2'-0 methyl or 2'-deoxy-2'-fluoro pyrimidine nucleotides and the purine nucleotides present in the antisense strand are 2'-O-methyl nucleotides or 2'-deoxy nucleotides. In other embodiments, the pyrimidine nucleotides in the sense strand are 2'-deoxy-2'-fluoro pyrimidine nucleotides and the purine nucleotides present in the sense strand are 2'-0-methyl or 2'-deoxy purine nucleotides.
[0154] In certain embodiments of the invention, all the pyrimidine nucleotides in the complementary region on the sense strand are 2'-deoxy-2'-fluoro pyrimidine nucleotides. In certain embodiments, all of the pyrimidine nucleotides in the complementary region of the antisense strand are 2'-deoxy-2'-fluoro pyrimidine nucleotides. In certain embodiments, all the purine nucleotides in the complementary region on the sense strand are 2'-deoxy purine nucleotides. In certain embodiments, all of the purines in the complementary region on the antisense strand are 2'-0-methyl purine nucleotides. In certain embodiments, all of the pyrimidine nucleotides in the complementary regions on the sense strand are 2'-deoxy-2'-fluoro pyrimidine nucleotides; all of the pyrimidine nucleotides in the complementary region of the antisense strand are 2'-deoxy-2'-fluoro pyrimidine nucleotides; all the purine nucleotides in the complementary region on the sense strand are 2'-deoxy purine nucleotides and all of the purines in the complementary region on the antisense strand are 2'-0-methyl purine nucleotides.
[0155] In some embodiments, at least 5 or more of the pyrimidine nucleotides in one or both stands are 2'-deoxy-2'-fluoro pyrimidine nucleotides. In some embodiments, at least 5 or more of the pyrimidine nucleotides in one or both stands are 2'-0-methyl pyrimidine nucleotides. In some embodiments, at least 5 or more of the purine nucleotides in one or both stands are 2' deoxy-2'-fluoro purine nucleotides In some embodiments, at least 5 or more of the purine nucleotides in one or both stands are 2'-0-methyl purine nucleotides.
[0156] In certain embodiments, the purines and pyrimidines are differentially modified at the 2'-sugar position (i.e., at least one purine has a different modification from at least one pyrimidine in the same or different strand at the 2'-sugar position). For example, in some instances, at least 5 or more of the pyrimidine nucleotides in one or both stands are 2'-deoxy-2' fluoro pyrimidine nucleotides and at least 5 or more purine nucleotides in one or both strands are 2'-O-methyl purine nucleotides. In other instances at least 5 or more of the pyrimidine nucleotides in one or both stands are 2'-0-methyl pyrimidine nucleotides and at least 5 or more purine nucleotides in one or both strands are 2'-deoxy-2'-fluoro purine nucleotides.
[0157] Further non-limiting examples of sense and antisense strands of such siNA molecules having various modifications and modifications patterns are shown in Figures 2 and 3.
[0158] Any of the above described modifications, or combinations thereof, including those in the references cited, can be applied to any of the siNA molecules of the invention.
[0159] The modified siNA molecules of the invention can comprise modifications at various locations within the siNA molecule. In some embodiments, the double-stranded siNA molecule of the invention comprises modified nucleotides at internal base paired positions within the siNA duplex. In other embodiments, a double-stranded siNA molecule of the invention comprises modified nucleotides at non-base paired or overhang regions of the siNA molecule. In yet other embodiments, a double-stranded siNA molecule of the invention comprises modified nucleotides at terminal positions of the siNA molecule. For example, such terminal regions include the 3' position and/or 5'-position of the sense and/or antisense strand or region of the siNA molecule. Additionally, any of the modified siNA molecules of the invention can have a modification in one or both oligonucleotide strands of the siNA duplex, for example in the sense strand, the antisense strand, or both strands. Moreover, with regard to chemical modifications of the siNA molecules of the invention, each strand of the double-stranded siNA molecules of the invention can have one or more chemical modifications, such that each strand comprises a different pattern of chemical modifications.
[0160] In certain embodiments each strand of a double-stranded siNA molecule of the invention comprises a different pattern of chemical modifications, such as any Stab modification chemistries described herein (see Table 8) or any combination thereof, i.e., different combinations of defined Stabilzation chemistry (Stab) sense and antisense strands. Further, non limiting examples of modification schemes that could give rise to different patterns of modifications are shown in Table 8. The stabilization chemistries referred to in Table 8 as Stab, can be combined in any combination of sense/antisense chemistries, such as Stab 7/8, Stab 7/11, Stab 8/8, Stab 18/8, Stab 18/11, Stab 12/13, Stab 7/13, Stab 18/13, Stab 7/19, Stab 8/19, Stab 18/19, Stab 7/20, Stab 8/20, Stab 18/20, Stab 7/32, Stab 8/32, or Stab 18/32 or any other combination of Stabilization chemistries.
[0161] In any of the siNAs of the invention, one or more (for example 1, 2, 3, 4 or 5) nucleotides at the 5'-end of the guide strand or guide region (also known as antisense strand or antisense region) of the siNA molecule are ribonucleotides.
[0162] In certain embodiments, the present invention provides a double-stranded short interfering nucleic acid (siNA) molecule that modulates the expression of HBV, wherein the siNA comprises a sense strand and an antisense strand; each strand is independently 15 to 30 nucleotides in length; and the antisense strand comprises at least 15 nucleotides having sequence complementary to any of:
5'- UCGUGGUGGACUUCUCUCA -3' (SEQ ID NO: 1); 5'- GUGGUGGACUUCUCUCAAU -3' (SEQ ID NO: 2); 5'- GCCGAUCCAUACUGCGGAA -3' (SEQ ID NO: 3); or 5'- CCGAUCCAUACUGCGGAAC -3' (SEQ ID NO: 4).
[0163] In some embodiments, the antisense strand of a siNA molecule of the invention comprises at least a 15 nucleotide sequence of:
5'-UGAGAGAAGUCCACCACGA-3'(SEQ ID NO: 452); 5'-AUUGAGAGAAGUCCACCAC-3'(SEQ ID NO: 453); 5'-UUCCGCAGUAUGGAUCGGC-3'(SEQ ID NO: 454); or
5'-GUUCCGCAGUAUGGAUCGG-3'(SEQ ID NO: 455).
[0164] In some embodiments, the sense strand of a siNA molecule of the invention comprises at least a 15 nucleotide sequence of:
5'- UCGUGGUGGACUUCUCUCA -3' (SEQ ID NO: 1); 5'- GUGGUGGACUUCUCUCAAU -3' (SEQ ID NO: 2); 5'- GCCGAUCCAUACUGCGGAA -3' (SEQ ID NO: 3); or 5'- CCGAUCCAUACUGCGGAAC -3' (SEQ ID NO: 4).
[0165] In some embodiments, a siNA molecule of the invention comprises any of:
5'-UCGUGGUGGACUUCUCUCA-3' (SEQ ID NO: 1) and 5' UGAGAGAAGUCCACCACGA -3'(SEQ ID NO: 452); or 5'-GUGGUGGACUUCUCUCAAU-3' (SEQ ID NO: 2); and 5' AUUGAGAGAAGUCCACCAC -3'(SEQ ID NO: 453); or 5'-GCCGAUCCAUACUGCGGAA-3' (SEQ ID NO: 3) and 5' UUCCGCAGUAUGGAUCGGC -3'(SEQ ID NO: 454); or 5'-CCGAUCCAUACUGCGGAAC-3' (SEQ ID NO: 4); and 5' GUUCCGCAGUAUGGAUCGG -3'(SEQ ID NO: 455).
[0166] Any of the above described modifications, or combinations thereof, including those in the references cited, can be applied to any of these embodiments.
[0167] In certain embodiments, the nucleotides of the at least a 15 nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 452, SEQ ID NO: 2, SEQ ID NO: 453, SEQ ID NO: 3, SEQ ID NO: 454, SEQ ID NO: 4, or SEQ ID NO:455 form a contiguous stretch of nucleotides.
[0168] In some embodiments, the siNA molecule can contain one or more nucleotide deletions, substitutions, mismatches and/or additions to the at least 15 nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 452, SEQ ID NO: 2, SEQ ID NO: 453, SEQ ID NO: 3, SEQ ID NO: 454, SEQ ID NO: 4, or SEQ ID NO:455; provided, however, that the siNA molecule maintains its activity, for example, to mediate RNAi. In a non-limiting example, the deletion, substitution, mismatch and/or addition can result in a loop or bulge, or alternately a wobble or other alternative (non Watson-Crick) base pair.
[0169] In certain embodiments of the invention, double-stranded siNA molecules are provided, wherein the molecule has a sense strand and an antisense strand and comprises the following formula (A):
B Nx 3 (N)x2 B -3' B (N)xI Nx 4 [Nlx5 -5' (A) wherein, the upper strand is the sense strand and the lower strand is the antisense strand of the double-stranded nucleic acid molecule; wherein the antisense strand comprises at least a 15 nucleotide sequence of SEQ ID NO: 452, SEQ ID NO: 453, SEQ ID NO: 454, or SEQ ID NO: 455, and the sense strand comprises a sequence having complementarity to the antisense strand; each N is independently a nucleotide which is unmodified or chemically modified or a non-nucleotide; each B is a terminal cap that is present or absent;
(N) represents overhanging nucleotides, each of which is independently unmodified or chemically modified;
[N] represents nucleotides that are ribonucleotides;
X1 and X2 are independently integers from 0 to 4;
X3 is an integer from 15 to 30;
X4 is an integer from 9 to 30; and
X5 is an integer from 0 to 6, provided that the sum of X4 and X5 is 15-30.
[0170] In certain embodiments, the nucleotides of the at least a 15 nucleotide sequence of SEQ ID NO: 452, SEQ ID NO: 453, SEQ ID NO: 454, or SEQ ID NO: 455 form a contiguous stretch of nucleotides.
[0171] In some embodiments, the siNA molecule of formula A can contain one or more nucleotide deletions, substitutions, mismatches and/or additions to the at least 15 nucleotide sequence of SEQ ID NO: 452, SEQ ID NO: 453, SEQ ID NO: 454, or SEQ ID NO: 455; provided, however, that the siNA molecule maintains its activity, for example, to mediate RNAi. In a non-limiting example, the deletion, substitution, mismatch and/or addition can result in a loop or bulge, or alternately a wobble or other alternative (non Watson-Crick) base pair.
[0172] In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) of formula (A); wherein
(a) one or more pyrimidine nucleotides in Nx4 positions are independently 2'-deoxy 2'-fluoro nucleotides, 2'-O-alkyl nucleotides, 2'-deoxy nucleotides, ribonucleotides, or any combination thereof;
(b) one or more purine nucleotides in Nx4 positions are independently 2'-deoxy-2' fluoro nucleotides, 2'-O-alkyl nucleotides, 2'-deoxy nucleotides, ribonucleotides, or any combination thereof;
(c) one or more pyrimidine nucleotides in Nx3 positions are independently 2'-deoxy 2'-fluoro nucleotides, 2'-O-alkyl nucleotides, 2'-deoxy nucleotides, ribonucleotides, or any combination thereof; and
(d) one or more purine nucleotides in Nx 3 positions are independently 2'-deoxy-2' fluoro nucleotides, 2'-O-alkyl nucleotides, 2'-deoxy nucleotides, ribonucleotides, or any combination thereof.
[0173] In certain embodiments, the invention features a double-stranded short interfering nucleic acid (siNA) molecule of formula (A); wherein (a) 1, 2, 3, 4, 5 or more pyrimidine nucleotides in Nx4 positions are 2'-deoxy-2' fluoro nucleotides;
(b) 1, 2, 3, 4, 5 or more purine nucleotides in Nx 4 positions are 2'-O-alkyl nucleotides;
(c) 1, 2, 3, 4, 5 or more pyrimidine nucleotides in Nx3 positions are 2'-deoxy-2' fluoro nucleotides; and
(d) 1, 2, 3, 4, 5 or more purine nucleotides in Nx 3 positions are 2'-deoxy nucleotides.
[0174] In certain embodiments, the invention features a double-stranded short interfering nucleic acid (siNA) molecule of formula (A); wherein
(a) 1, 2, 3, 4, 5 or more pyrimidine nucleotides in Nx4 positions are 2'-O-alkyl nucleotides; (b) 1, 2, 3, 4, 5 or more purine nucleotides in Nx4 positions are ribonucleotides;
(c) 1, 2, 3, 4, 5 or more pyrimidine nucleotides in Nx 3 positions are 2'-O-alkyl nucleotides; and
(d) 1, 2, 3, 4, 5 or more purine nucleotides in Nx3 positions are ribonucleotides.
[0175] In certain embodiments, the invention features a double-stranded short interfering nucleic acid (siNA) molecule of formula (A); wherein
(a) 1, 2, 3, 4, 5 or more pyrimidine nucleotides in Nx4 positions are 2'-deoxy-2' fluoro nucleotides; (b) 1, 2, 3, 4, 5 or more purine nucleotides in Nx4 positions are 2'-O-alkyl nucleotides;
(c) 1, 2, 3, 4, 5 or more pyrimidine nucleotides in Nx 3 positions are 2'-O-alkyl nucleotides; and
(d) 1, 2, 3, 4, 5 or more purine nucleotides in Nx3 positions are 2'-deoxy-2'-fluoro nucleotides.
[0176] In certain embodiments, the invention features a double-stranded short interfering nucleic acid (siNA) molecule of formula (A) further comprising one or more phosphorothioate internucleotide linkages.
[0177] In some embodiments, siNA molecules having formula A comprise a terminal phosphate group at the 5'-end of the antisense strand or antisense region of the nucleic acid molecule.
[0178] In various embodiments, siNA molecules having formula A comprise X5 = 0, 1, 2, or 3; each X1 and X2 = 1 or 2; X3 = 18, 19, 20, 21, 22, or 23, and X4 = 17, 18, 19, 20, 21, 22, or 23.
[0179] In certain embodiments, siNA molecules having formula A comprise X5 = 3. In other embodiments siNA molecules having formula A comprise X5 = 0.
[0180] In certain embodiments, siNA molecules having formula A comprise X= 2 and X2 = 2.
[0181] In various embodiments, siNA molecules having formula A comprise X5= 0, X =
2, and X2 = 2. In other embodiments, siNA molecules having formula A comprise X5 = 3, X1 =2, and X2= 2.
[0182] In one specific embodiment, an siNA molecule having formula A comprises X5 = 3; each X1 and X2 = 2; X3 = 19, and X4 = 16..
[0183] In another specific embodiment, an siNA molecule having formula A comprises X5 = ; each X1 and X2 = 2; X3 = 19, and X4 = 19.
[0184] In certain embodiments, siNA molecules having formula A comprise caps (B) at the 3' and 5' ends of the sense strand or sense region.
[0185] In certain embodiments, siNA molecules having formula A comprise caps (B) at the 3'-end of the antisense strand or antisense region.
[0186] In various embodiments, siNA molecules having formula A comprise caps (B) at the 3' and 5' ends of the sense strand or sense region and caps (B) at the3'-end of the antisense strand or antisense region.
[0187] In yet other embodiments, siNA molecules having formula A comprise caps (B) only at the 5'-end of the sense (upper) strand of the double-stranded nucleic acid molecule.
[0188] In some embodiments, siNA molecules having formula A further comprise one or more phosphorothioate internucleotide linkages between the nucleotides. In certain embodiments, siNA molecules having formula A comprise one or more phosphorothioate internucleotide linkages between the first terminal (N) and the adjacent nucleotide on the 3'end of the sense strand, antisense strand, or both sense strand and antisense strands of the nucleic acid molecule. For example, a double-stranded nucleic acid molecule can comprise X1 and/or X2 = 2 having overhanging nucleotide positions with a phosphorothioate internucleotide linkage, e.g., (NsN) where "s" indicates phosphorothioate.
[0189] In some embodiments, one or more of the nucleotides of siNA molecules having formula A have a universal base.
[0190] In certain embodiments, siNA molecules having formula A have at position 14 from the 5'-end of the antisense strand a ribonucleotide when the nucleotide at that position 14 is a purine. In other embodiments, siNA molecules having formula A have at position 14 from the
'-end of the antisense strand a ribonucleotide, a 2'-deoxy-2'-fluoro nucleotide or a 2'--methyl nucleotide when the nucleotide at that position 14 is a pyrimidine nucleotide.
[0191] In some embodiments, siNA molecules having formula A comprises (N) nucleotides in the antisense strand (lower strand) that are complementary to nucleotides in a HBV target polynucleotide sequence, which also has complementarity to the N and [N] nucleotides of the antisense (lower) strand.
[0192] Any of the above described modifications, or combinations thereof, discussed above as applicable to siNAs of the invention, including those in the references cited, can be applied to any of the embodiments to siNA molecules having formula A.
C. Generation/Synthesis of siNA Molecules
[0193] The siNAs of the invention can be obtained using a number of techniques known to those of skill in the art. For example the siNA can be chemically synthesized or may be encoded by plasmid (e.g., transcribed as sequences that automatically fold into duplexes with hairpin loops.). siNA can also be generated by cleavage of longer dsRNA (e.g., dsRNA greater than about 25 nucleotides in length) by the E coli RNase II or Dicer. These enzymes process the dsRNA into biologically active siNA (see, e.g., Yang et al., PNAS USA 99:9942-9947 (2002); Calegari et al. PNAS USA 99:14236 (2002) Byron et al. Ambion Tech Notes; 10 (1):4-6 (2009); Kawaski et al., Nucleic Acids Res., 31:981-987 (2003), Knight and Bass, Science, 293:2269 2271 (2001) and Roberston et al., J. Biol. Chem 243:82(1969).
1. Chemical Synthesis
[0194] Preferably, siNA of the invention are chemically synthesized. Oligonucleotides (e.g., certain modified oligonucleotides or portions of oligonucleotides lacking ribonucleotides) are synthesized using protocols known in the art, for example as described in Caruthers et al., 1992, Methods in Enzymology 211, 3-19, Thompson et al., International PCT Publication No. WO 99/54459, Wincott et al., 1995, Nucleic Acids Res. 23, 2677-2684, Wincott et al., 1997, Methods Mol. Bio., 74, 59, Brennan et al., 1998, Biotechnol Bioeng., 61, 33-45, and Brennan, U.S. Pat. No. 6,001,311. The synthesis of oligonucleotides makes use of common nucleic acid protecting and coupling groups, such as dimethoxytrityl at the 5'-end, and phosphoramidites at the 3'-end.
[0195] siNA molecules without modifications are synthesized using procedures as described in Usman et al., 1987, J. Am. Chem. Soc., 109, 7845; Scaringe et al., 1990, Nucleic Acids Res., 18, 5433. These syntheses makes use of common nucleic acid protecting and coupling groups, such as dimethoxytrityl at the 5'-end, and phosphoramidites at the 3'-end that can be used for certain siNA molecules of the invention.
[0196] In certain embodiments, the siNA molecules of the invention are synthesized, deprotected, and analyzed according to methods described in U.S. Patent Nos. 6,995,259, 6,686,463, 6,673,918, 6,649,751, 6,989,442, and U.S. Patent Application No. 10/190,359.
[0197] In a non-limiting synthesis example, small scale syntheses are conducted on a 394 Applied Biosystems, Inc. synthesizer using a 0.2 pmol scale protocol with a 2.5 min coupling step for 2'-0-methylated nucleotides and a 45 second coupling step for 2'-deoxy nucleotides or 2'-deoxy-2'-fluoro nucleotides. Table 9 outlines the amounts and the contact times of the reagents used in the synthesis cycle.
[0198] Alternatively, the siNA molecules of the present invention can be synthesized separately and joined together post-synthetically, for example, by ligation (Moore et al., 1992, Science 256, 9923; Draper et al., International PCT Publication No. WO 93/23569; Shabarova et al., 1991, Nucleic Acids Research 19, 4247; Bellon et al., 1997, Nucleosides & Nucleotides, 16, 951; Bellon et al., 1997, Bioconjugate Chem. 8, 204), or by hybridization following synthesis and/or deprotection.
[0199] Various siNA molecules of the invention can also be synthesized using the teachings of Scaringe et al., US Patent Nos. 5,889,136; 6,008,400; and 6,111,086.
2. Vector Expression
[0200] Alternatively, siNA molecules of the invention that interact with and down-regulate gene encoding target HBV molecules can be expressed and delivered from transcription units (see for example Couture et al., 1996, TIG., 12, 510) inserted into DNA or RNA vectors. The recombinant vectors can be DNA plasmids or viral vectors. siNA expressing viral vectors can be constructed based on, but not limited to, adeno-associated virus, retrovirus, adenovirus, or alphavirus.
[0201] In some embodiments, pol III based constructs are used to express nucleic acid molecules of the invention. Transcription of the siNA molecule sequences can be driven from a promoter for eukaryotic RNA polymerase I (pol I), RNA polymerase II (pol II), or RNA polymerase III (pol III). (see for example, Thompson, U.S. Patent. Nos. 5,902,880 and 6,146,886). (See also, Izant and Weintraub, 1985, Science, 229, 345; McGarry and Lindquist, 1986, Proc. Nat. Acad. Sci., USA 83, 399; Scanlon et al., 1991, Proc. Nat. Acad. Sci. USA, 88, 10591-5; Kashani-Sabet et al., 1992, Antisense Res. Dev., 2, 3-15; Dropulic et al., 1992, J. Virol., 66, 1432-41; Weerasinghe et al., 1991, J. Virol., 65, 5531-4; Ojwang et al., 1992, Proc. Nat/. Acad. Sci. USA, 89, 10802-6; Chen et al., 1992, Nucleic Acids Res., 20, 4581-9; Sarver et al., 1990 Science, 247, 1222-1225; Thompson et al., 1995, Nucleic Acids Res., 23, 2259; Good et al., 1997, Gene Therapy, 4, 45. Transcripts from pol II or pol III promoters are expressed at high levels in all cells; the levels of a given pol II promoter in a given cell type depends on the nature of the gene regulatory sequences (enhancers, silencers, etc.) present nearby. Prokaryotic RNA polymerase promoters are also used, providing that the prokaryotic RNA polymerase enzyme is expressed in the appropriate cells (Elroy-Stein and Moss, 1990, Proc. Nat. Acad. Sci. USA, 87, 6743-7; Gao and Huang 1993, Nucleic Acids Res., 21, 2867-72; Lieber et al., 1993, Methods Enzymol., 217, 47-66; Zhou et al., 1990, Mol. Cell. Biol., 10, 4529-37). Several investigators have demonstrated that nucleic acid molecules expressed from such promoters can function in mammalian cells (e.g. Kashani-Sabet et al., 1992, Antisense Res. Dev., 2, 3-15; Ojwang et al., 1992, Proc. Natl. Acad. Sci. U S A, 89, 10802-6; Chen et al., 1992, Nucleic Acids Res., 20, 4581 9; Yu et al., 1993, Proc. Natl. Acad. Sci. U S A, 90, 6340-4; L'Huillier et al., 1992, EMBO J., 11, 4411-8; Lisziewicz et al., 1993, Proc. Natl. Acad. Sci. U. S. A, 90, 8000-4; Thompson et al., 1995, Nucleic Acids Res., 23, 2259; Sullenger & Cech, 1993, Science, 262, 1566). More specifically, transcription units such as the ones derived from genes encoding U6 small nuclear (snRNA), transfer RNA (tRNA) and adenovirus VA RNA are useful in generating high concentrations of desired RNA molecules such as siNA in cells (Thompson et al., supra; Couture and Stinchcomb, 1996, supra; Noonberg et al., 1994, Nucleic Acid Res., 22, 2830; Noonberg et al., U.S. Pat. No. 5,624,803; Good et al., 1997, Gene Ther., 4, 45; Beigelman et al., International PCT Publication No. WO 96/18736. The above siNA transcription units can be incorporated into a variety of vectors for introduction into mammalian cells, including but not restricted to, plasmid DNA vectors, viral DNA vectors (such as adenovirus or adeno-associated virus vectors), or viral RNA vectors (such as retroviral or alphavirus vectors) (for a review see Couture and Stinchcomb, 1996, supra).
[0202] Vectors used to express the siNA molecules of the invention can encode one or both strands of an siNA duplex, or a single self-complementary strand that self hybridizes into an siNA duplex. The nucleic acid sequences encoding the siNA molecules of the instant invention can be operably linked in a manner that allows expression of the siNA molecule (see for example Paul et al., 2002, Nature Biotechnology, 19, 505; Miyagishi and Taira, 2002, Nature Biotechnology, 19, 497; Lee et al., 2002, Nature Biotechnology, 19, 500; and Novina et al., 2002, Nature Medicine, advance online publication doi:10.1038/nm725).
D. Carrier/Delivery Systems
[0203] The siNA molecules of the invention are added directly, or can be complexed with cationic lipids, packaged within liposomes, or as a recombinant plasmid or viral vectors which express the siNA molecules, or otherwise delivered to target cells or tissues. Methods for the delivery of nucleic acid molecules are described in Akhtar et al., 1992, Trends Cell Bio., 2, 139; Delivery Strategiesfor Antisense Oligonucleotide Therapeutics, ed. Akhtar, 1995, Maurer et al., 1999, Mol. Membr. Biol., 16,129-140; Hofland and Huang, 1999, Handb. Exp. Pharmacol., 137, 165-192; and Lee et al., 2000, ACS Symp. Ser., 752, 184-192. Beigelman et al., U.S. Pat. No. 6,395,713 and Sullivan et al., PCT WO 94/02595 further describe the general methods for delivery of nucleic acid molecules. These protocols can be utilized for the delivery of virtually any nucleic acid molecule. Nucleic acid molecules can be administered to cells by a variety of methods known to those of skill in the art, including, but not restricted to, encapsulation in liposomes, by iontophoresis, or by incorporation into other vehicles, such as biodegradable polymers, hydrogels, cyclodextrins (see for example, Gonzalez et al., 1999, Bioconjugate Chem., , 1068-1074; Wang et al., International PCT Publication Nos. WO 03/47518 and WO 03/46185), poly(lactic-co-glycolic)acid (PLGA) and PLCA microspheres (see for example US Patent 6,447,796 and US Patent Application Publication No. US 2002130430), biodegradable nanocapsules, and bioadhesive microspheres, or by proteinaceous vectors (O'Hare and Normand, International PCT Publication No. WO 00/53722).
[0204] In one aspect, the present invention provides carrier systems containing the siNA molecules described herein. In some embodiments, the carrier system is a lipid-based carrier system, cationic lipid, or liposome nucleic acid complexes, a liposome, a micelle, a virosome, a lipid nanoparticle or a mixture thereof. In other embodiments, the carrier system is a polymer based carrier system such as a cationic polymer-nucleic acid complex. In additional embodiments, the carrier system is a cyclodextrin-based carrier system such as a cyclodextrin polymer-nucleic acid complex. In further embodiments, the carrier system is a protein-based carrier system such as a cationic peptide-nucleic acid complex. Preferably, the carrier system is a lipid nanoparticle ("LNP") formulation.
[0205] In certain embodiments, the siNA molecules of the invention are formulated with a lipid nanoparticle composition such as is described in U.S. Patent Application Nos. 11/353,630, 11/586,102, 61/189,295, 61/204,878, 61/235,476, 61/249,807, 61/298,022, 61/351373, 61/347640, 61/345754, 61/322054, 12/640342, and 12/617079, and PCT Applications Nos.
PCT/US10/020013 and PCT/US09/053336. In certain embodiments, the siNA is formulated with components and ratios described and exemplified in Example 6 herein (see also Tables 10 & 11).
[0206] In various embodiments, lipid nanoparticle formulations described in Table 10 are applied to any siNA molecule or combination of siNA molecules herein. In some embodiments, the invention features a composition comprising an siNA molecule of the invention formulated as a composition including any of cationic lipid compound numbers 1-46 or a combination thereof.
[0207] In certain other embodiments, the invention features a composition comprising an siNA molecule of the invention formulated with any of the cationic lipid formulations described in U.S. Patent Application Nos. 61/189,295, 61/204,878, 61/235,476, 61/249,807, and 61/298,022.
[0208] In other embodiments, the invention features conjugates and/or complexes of siNA molecules of the invention. Such conjugates and/or complexes can be used to facilitate delivery of siNA molecules into a biological system, such as a cell. The conjugates and complexes provided by the instant invention can impart therapeutic activity by transferring therapeutic compounds across cellular membranes, altering the pharmacokinetics, and/or modulating the localization of nucleic acid molecules of the invention. Non-limiting, examples of such conjugates are described in U.S. Publication Nos. US2008/0152661 Al and US 2004/0162260 Al (e.g., CDM-LBA, CDM-Pip-LBA, CDM-PEG, CDM-NAG, etc.) and U.S. Patent Application Nos. 10/427,160 10/201,394, 61/322422, and 61/315223; and U.S. Patent Nos. 6,528,631; 6,335,434; 6,235,886; 6,153,737; 5,214,136; and 5,138,045.
[0209] In various embodiments, polyethylene glycol (PEG) can be covalently attached to siNA compounds of the present invention. The attached PEG can be any molecular weight, preferably from about 100 to about 50,000 daltons (Da).
[0210] In yet other embodiments, the invention features compositions or formulations comprising surface-modified liposomes containing poly (ethylene glycol) lipids (PEG-modified, or long-circulating liposomes or stealth liposomes) and siNA molecules of the invention, such as is disclosed in for example, International PCT Publication No. WO 96/10391; Ansell et al., International PCT Publication No. WO 96/10390; Holland et al., International PCT Publication No. WO 96/10392.
[0211] In some embodiments, the siNA molecules of the invention can also be formulated or complexed with polyethyleneimine and derivatives thereof, such as polyethyleneimine polyethyleneglycol-N-acetylgalactosamine (PEI-PEG-GAL) or polyethyleneimine polyethyleneglycol-tri-N-acetylgalactosamine (PEI-PEG-triGAL) derivatives. In one embodiment, the nucleic acid molecules of the invention are formulated as described in U.S. Patent Application Publication No. 20030077829.
[0212] In other embodiments, siNA molecules of the invention are complexed with membrane disruptive agents such as those described in U.S. Patent Application Publication No. 20010007666. In still other embodiments, the membrane disruptive agent or agents and the siNA molecule are also complexed with a cationic lipid or helper lipid molecule, such as those lipids described in U.S. Patent No. 6,235,310.
[0213] In certain embodiments, siNA molecules of the invention are complexed with delivery systems as described in U.S. Patent Application Publication Nos. 2003077829;
20050287551; 20050164220; 20050191627; 20050118594; 20050153919; 20050085486; and 20030158133; and International PCT Publication Nos. WO 00/03683 and WO 02/087541.
[0214] In some embodiments, a liposomal formulation of the invention comprises an siNA molecule of the invention (e.g., siNA) formulated or complexed with compounds and compositions described in U.S. Patent Nos. 6,858,224; 6,534,484; 6,287,591; 6,835,395; 6,586,410; 6,858,225; 6,815,432; 6,586,001; 6,120,798; 6,977,223; 6,998,115; 5,981,501; ,976,567; 5,705,385; and U.S. Patent Application Publication Nos. 2006/0019912; 2006/0019258; 2006/0008909; 2005/0255153; 2005/0079212; 2005/0008689; 2003/0077829, 2005/0064595, 2005/0175682, 2005/0118253; 2004/0071654; 2005/0244504; 2005/0265961 and 2003/0077829.
[0215] Alternatively, recombinant plasmids and viral vectors, as discussed above, which express siNAs of the invention can be used to deliver the molecules of the invention. Delivery of siNA molecule expressing vectors can be systemic, such as by intravenous or intra-muscular administration, by administration to target cells ex-planted from a subject followed by reintroduction into the subject, or by any other means that would allow for introduction into the desired target cell (for a review see Couture et al., 1996, TIG., 12, 510). Such recombinant plasmids can also be administered directly or in conjunction with a suitable delivery reagents, including, for example, the Mirus Transit LTI lipophilic reagent; lipofectin; lipofectamine; cellfectin; polycations (e.g., polylysine) or liposomes lipid-based carrier system, cationic lipid, or liposome nucleic acid complexes, a micelle, a virosome, a lipid nanoparticle.
E. Kits
[0216] The present invention also provides nucleic acids in kit form. The kit may comprise a container. The kit typically contains a nucleic acid of the invention with instructions for its administration. In certain instances, the nucleic acids may have a targeting moiety attached. Methods of attaching targeting moieties (e.g. antibodies, proteins) are known to those of skill in the art. In certain instances, the nucleic acids are chemically modified. In other embodiments, the kit contains more than one siNA molecule of the invention. The kits may comprise an siNA molecule of the invention with a pharmaceutically acceptable carrier or diluent. The kits may further comprise excipients.
F. Therapeutic Uses/Pharmaceutical Compositions
[0217] The present body of knowledge in HBV research indicates the need for methods to assay HBV activity and for compounds that can regulate HBV expression for research, diagnostic, and therapeutic use. As described infra, the nucleic acid molecules of the present invention can be used in assays to diagnose disease state related of HBV levels. In addition, the nucleic acid molecules and pharmaceutical compositions can be used to treat disease states related to HBV RNA levels.
1. Disease States Associated with HBV
[0218] Particular disease states that can be associated with HBV expression modulation include liver disease and cancer. Non-limiting examples of such diseases include cirrhosis of the liver and heptocellular carcinoma.
[0219] It is understood that the siNA molecules of the invention can degrade the target HBV mRNA (and thus inhibit the diseases stated above). Inhibition of a disease can be evaluated by directly measuring the progress of the disease in a subject. It can also be inferred through observing a change or reversal in a condition associated with the disease. Additionally, the siNA molecules of the invention can be used as a prophylaxis. Thus, the use of the nucleic acid molecules and pharmaceutical compositions of the invention can be used to ameliorate, treat, prevent, and/or cure these diseases and others associated with regulation of HBV gene expression.
2. Pharmaceutical Compositions
[0220] The siNA molecules of the instant invention provide useful reagents and methods for a variety of therapeutic, prophylactic, cosmetic, veterinary, diagnostic, target validation, genomic discovery, genetic engineering, and pharmacogenomic applications.
a. Formulations
[0221] Thus, the present invention, in one aspect, also provides for pharmaceutical compositions of the siNA molecules described, i.e., compositions in a pharmaceutically acceptable carrier or diluent. These pharmaceutical compositions include salts, esters, or salts of such esters, of the above compounds, e.g., acid addition salts, for example, salts of hydrochloric, hydrobromic, hydroiodic, acetic acid, and benzene sulfonic acid. Other salts include for example, sodium, potassium, manganese, ammonium, and calcium salts. These formulations or compositions can comprise a pharmaceutically acceptable carrier or diluent as is generally known in the art.
[0222] In one embodiment, the invention features a pharmaceutical composition comprising an siNA molecule comprising at least a 15 nucleotide sequence of SEQ ID NO: 1. In another embodiment, the invention features a pharmaceutical composition comprising an siNA molecule comprising at least a 15 nucleotide sequence of SEQ ID NO: 452. In yet another embodiment, the invention features a pharmaceutical composition comprising an siNA molecule comprising at least a 15 nucleotide sequence of SEQ ID NO: 2. In still another embodiment, the invention features a pharmaceutical composition comprising an siNA molecule comprising at least a 15 nucleotide sequence of SEQ ID NO: 453. In another embodiment, the invention features a pharmaceutical composition comprising an siNA molecule comprising at least a 15 nucleotide sequence of SEQ ID NO: 3. In another embodiment, the invention features a pharmaceutical composition comprising an siNA molecule comprising at least a 15 nucleotide sequence of SEQ ID NO: 454. In another embodiment, the invention features a pharmaceutical composition comprising an siNA molecule comprising at least a 15 nucleotide sequence of SEQ ID NO: 4. In yet another embodiment, the invention features a pharmaceutical composition comprising an siNA molecule comprising at least a 15 nucleotide sequence of SEQ ID NO: 455. In still another embodiment, the invention features a pharmaceutical composition comprising an siNA molecule comprising formula (A).
[0223] The siNA molecules of the invention are preferably formulated as pharmaceutical compositions prior to administering to a subject, according to techniques known in the art. Pharmaceutical compositions of the present invention are characterized as being at least sterile and pyrogen-free. Methods for preparing pharmaceutical compositions of the invention are within the skill in the art for example as described in Remington's PharmaceuticalScience, 17t ed., Mack Publishing Company, Easton, Pa. (1985).
[0224] In some embodiments, pharmaceutical compositions of the invention (e.g. siNA and/or LNP formulations thereof) further comprise conventional pharmaceutical excipients and/or additives. Suitable pharmaceutical excipients include preservatives, flavoring agents, stabilizers, antioxidants, osmolality adjusting agents, buffers, and pH adjusting agents. Suitable additives include physiologically biocompatible buffers (e.g., trimethylamine hydrochloride), addition of chelants (such as, for example, DTPA or DTPA-bisamide) or calcium chelate complexes (as for example calcium DTPA, CaNaDTPA-bisamide), or, optionally, additions of calcium or sodium salts (for example, calcium chloride, calcium ascorbate, calcium gluconate or calcium lactate). In addition, antioxidants and suspending agents can be used.
[0225] Non-limiting examples of various types of formulations for local administration include ointments, lotions, creams, gels, foams, preparations for delivery by transdermal patches, powders, sprays, aerosols, capsules or cartridges for use in an inhaler or insufflator or drops (for example eye or nose drops), solutions/suspensions for nebulization, suppositories, pessaries, retention enemas and chewable or suckable tablets or pellets (for example for the treatment of aphthous ulcers) or liposome or microencapsulation preparations.
[0226] Ointments, creams and gels, can, for example, can be formulated with an aqueous or oily base with the addition of suitable thickening and/or gelling agent and/or solvents. Non limiting examples of such bases can thus, for example, include water and/or an oil such as liquid paraffin or a vegetable oil such as arachis oil or castor oil, or a solvent such as polyethylene glycol. Various thickening agents and gelling agents can be used depending on the nature of the base. Non-limiting examples of such agents include soft paraffin, aluminum stearate, cetostearyl alcohol, polyethylene glycols, woolfat, beeswax, carboxypolymethylene and cellulose derivatives, and/or glyceryl monostearate and/or non-ionic emulsifying agents.
[0227] In one embodiment lotions can be formulated with an aqueous or oily base and will in general also contain one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents or thickening agents.
[0228] In one embodiment powders for external application can be formed with the aid of any suitable powder base, for example, talc, lactose or starch. Drops can be formulated with an aqueous or non-aqueous base also comprising one or more dispersing agents, solubilizing agents, suspending agents or preservatives.
[0229] Compositions intended for oral use can be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions can contain one or more such sweetening agents, flavoring agents, coloring agents or preservative agents in order to provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients that are suitable for the manufacture of tablets. These excipients can be, for example, inert diluents; such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, corn starch, or alginic acid; binding agents, for example starch, gelatin or acacia; and lubricating agents, for example magnesium stearate, stearic acid or talc. The tablets can be uncoated or they can be coated by known techniques. In some cases such coatings can be prepared by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monosterate or glyceryl distearate can be employed.
[0230] Formulations for oral use can also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example peanut oil, liquid paraffin or olive oil.
[0231] Aqueous suspensions contain the active materials in a mixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydropropyl-methylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents can be a naturally-occurring phosphatide, for example, lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate; or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions can also contain one or more preservatives, for example ethyl, or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.
[0232] Oily suspensions can be formulated by suspending the active ingredients in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions can contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents and flavoring agents can be added to provide palatable oral preparations. These compositions can be preserved by the addition of an anti oxidant such as ascorbic acid
[0233] Pharmaceutical compositions of the invention can also be in the form of oil-in-water emulsions. The oily phase can be a vegetable oil or a mineral oil or mixtures of these. Suitable emulsifying agents can be naturally-occurring gums, for example gum acacia or gum tragacanth, naturally-occurring phosphatides, for example soy bean, lecithin, and esters or partial esters derived from fatty acids and hexitol, anhydrides, for example sorbitan monooleate, and condensation products of the said partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate. The emulsions can also contain sweetening and flavoring agents.
[0234] Syrups and elixirs can be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol, glucose or sucrose. Such formulations can also contain a demulcent, a preservative and flavoring and coloring agents. The pharmaceutical compositions can be in the form of a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents that have been mentioned above. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parentally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.
[0235] The nucleic acid molecules of the invention can also be administered in the form of suppositories, e.g., for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials include cocoa butter and polyethylene glycols.
[0236] Nucleic acid molecules of the invention can be administered parenterally in a sterile medium. The drug, depending on the vehicle and concentration used, can either be suspended or dissolved in the vehicle. Advantageously, adjuvants such as local anesthetics, preservatives and buffering agents can be dissolved in the vehicle.
[0237] In other embodiments, the siNA and LNP compositions and formulations provided herein for use in pulmonary delivery further comprise one or more surfactants. Suitable surfactants or surfactant components for enhancing the uptake of the compositions of the invention include synthetic and natural as well as full and truncated forms of surfactant protein A, surfactant protein B, surfactant protein C, surfactant protein D and surfactant Protein E, di saturated phosphatidylcholine (other than dipalmitoyl), dipalmitoylphosphatidylcholine, phosphatidylcholine, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, phosphatidylserine; phosphatidic acid, ubiquinones, lysophosphatidylethanolamine, lysophosphatidylcholine,palmitoyl-lysophosphatidylcholine,dehydroepiandrosterone,dolichols, sulfatidic acid, glycerol-3-phosphate, dihydroxyacetone phosphate, glycerol, glycero-3 phosphocholine, dihydroxyacetone, palmitate, cytidine diphosphate (CDP) diacylglycerol, CDP choline, choline, choline phosphate; as well as natural and artificial lamellar bodies which are the natural carrier vehicles for the components of surfactant, omega-3 fatty acids, polyenic acid, polyenoic acid, lecithin, palmitinic acid, non-ionic block copolymers of ethylene or propylene oxides, polyoxypropylene, monomeric and polymeric, polyoxyethylene, monomeric and polymeric, poly (vinyl amine) with dextran and/or alkanoyl side chains, Brij 35, Triton X-100 and synthetic surfactants ALEC, Exosurf, Survan and Atovaquone, among others. These surfactants can be used either as single or part of a multiple component surfactant in a formulation, or as covalently bound additions to the 5' and/or 3' ends of the nucleic acid component of a pharmaceutical composition herein.
b. Combinations
[0238] The siNAs and pharmaceutical formulations according to the invention can be administered to a subject alone or used in combination with or include one or more other therapeutic agents, for example, antiviral or anticancer agents. Thus, combinations of the presently disclosed compounds with other antiviral or anti-cancer or chemotherapeutic agents are within the scope of the invention
[0239] Examples of anti-cancer or chemotherapeutic agents can be found in Cancer Principles and Practice of Oncology by V.T. Devita and S. Hellman (editors), 6" edition (February 15, 2001), Lippincott Williams & Wilkins Publishers. A person of ordinary skill in the art would be able to discern which combinations of agents would be useful based on the particular characteristics of the drugs and the cancer involved. Such anti-cancer agents include, but are not limited to, the following: estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic/cytostatic agents, antiproliferative agents, prenyl-protein transferase inhibitors, HMG-CoA reductase inhibitors and other angiogenesis inhibitors, inhibitors of cell proliferation and survival signaling, apoptosis inducing agents and agents that interfere with cell cycle checkpoints. The siNAs of the invention are also useful in combination with any therapeutic agent used in the treatment of HCC, for example, but not limitation sorafenib.
[0240] In a further embodiment, therefore, the invention provides a combination comprising an siNA molecule of the invention, such as for example, but not limitation, an siNA molecule comprising at least a 15 nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 1210, SEQ ID NO: 2, SEQ ID NO: 1211, SEQ ID NO: 3, SEQ ID NO: 1212, SEQ ID NO: 4, or SEQ ID NO:1213; or formula (A) or a pharmaceutically acceptable salt, solvate or physiologically functional derivative thereof together with one or more antiviral agents. In some embodiments, the antiviral is another HBV inhibitor.
[0241] In certain embodiments, the instant siNA molecules of the invention are useful in combination with known HBV antiviral agents including the following: Lamivudine (2',3' dideoxy-3'-thiacytidine, commonly called 3TC), Pegnterferon-alpha2a, tenofovir, entecavir, telbivudine, adefovir.
[0242] Also, the invention provides a combination comprising an siNA molecule of the invention, such as for example, but not limitation, an siNA molecule comprising at least a 15 nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 452, SEQ ID NO: 2, SEQ ID NO: 453, SEQ ID NO: 3, SEQ ID NO: 454, SEQ ID NO: 4, or SEQ ID NO:455; or formula (A) or a pharmaceutically acceptable salt, solvate or physiologically functional derivative thereof together with one or more anti-cancer or chemotherapeutic agents.
[0243] In certain embodiments, the instant siNA molecules of the invention are useful in combination with known anti-cancer agents including the following: estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic agents, antiproliferative agents, prenyl-protein transferase inhibitors, HMG-CoA reductase inhibitors, HIV protease inhibitors, reverse transcriptase inhibitors, and other angiogenesis inhibitors.
[0244] Examples of estrogen receptor modulators that can be used in combination with the compounds of the invention include, but are not limited to, tamoxifen, raloxifene, idoxifene, LY353381, LY117081, toremifene, fulvestrant, 4-[7-(2,2-dimethyl-1-oxopropoxy-4-methyl-2
[4-[2-(1-piperidinyl)ethoxy]phenyl]-2H-1-benzopyran-3-yl]-phenyl-2,2-dimethylpropanoate, 4,4'-dihydroxybenzophenone-2,4-dinitrophenyl-hydrazone, and SH646.
[0245] Examples of androgen receptor modulators that can be used in combination with the compounds of the invention include, but are not limited to, finasteride and other 5a-reductase inhibitors, nilutamide, flutamide, bicalutamide, liarozole, and abiraterone acetate.
[0246] Examples of such retinoid receptor modulators that can be used in combination with the compounds of the invention include, but are not limited to, bexarotene, tretinoin, 13-cis retinoic acid, 9-cis-retinoic acid, c-difluoromethylomithine, ILX23-7553, trans-N-(4' hydroxyphenyl) retinamide, and N-4-carboxyphenyl retinamide.
[0247] Examples of cytotoxic agents that can be used in combination with the compounds of the invention include, but are not limited to, sertenef, cachectin, ifosfamide, tasonermin, lonidamine, carboplatin, altretamine, prednimustine, dibromodulcitol, ranimustine, fotemustine, nedaplatin, oxaliplatin, temozolomide, heptaplatin, estramustine, improsulfan tosilate, trofosfamide, nimustine, dibrospidium chloride, pumitepa, lobaplatin, satraplatin, profiromycin, cisplatin, irofulven, dexifosfamide, cis-aminedichloro(2-methyl-pyridine)platinum, benzylguanine, glufosfamide, GPX100, (trans, trans, trans)-bis-mu-(hexane-1,6-diamine)-mu
[diamine-platinum(II)]bis[diamine(chloro)platinum (II)]tetrachloride, diarizidinylspermine, arsenic trioxide, 1-(11-dodecylamino-10-hydroxyundecyl)-3,7-dimethylxanthine, zorubicin, idarubicin, daunorubicin, bisantrene, mitoxantrone, pirarubicin, pinafide, valrubicin, amrubicin, antineoplaston, 3'-deamino-3'-morpholino-13-deoxo-10-hydroxycarminomycin, annamycin, galarubicin, elinafide, MEN10755, and 4-demethoxy-3-deamino-3-aziridinyl-4 methylsulphonyl-daunorubicin (see WO 00/50032).
[0248] An example of a hypoxia activatable compound that can be used in combination with the compounds of the invention is tirapazamine.
[0249] Examples of proteasome inhibitors that can be used in combination with the compounds of the invention include, but are not limited to, lactacystin and bortezomib.
[0250] Examples of microtubule inhibitors/microtubule-stabilising agents that can be used in combination with the compounds of the invention include, but are not limited to, paclitaxel, vindesine sulfate, 3',4'-didehydro-4'-deoxy-8'-norvincaleukoblastine, docetaxol, rhizoxin, dolastatin, mivobulin isethionate, auristatin, cemadotin, RPR109881, BMS184476, vinflunine, cryptophycin, 2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methoxyphenyl) benzene sulfonamide, anhydrovinblastine, N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-prolyl-L-proline-t butylamide, TDX258, the epothilones (see for example U.S. Pat. Nos. 6,284,781 and 6,288,237) and BMS188797.
[0251] Some examples of topoisomerase inhibitors that can be used in combination with the compounds of the invention include, but are not limited to, are topotecan, hycaptamine, irinotecan, rubitecan, 6-ethoxypropionyl-3',4'-O-exo-benzylidene-chartreusin, 9-methoxy-N,N dimethyl-5-nitropyrazolo[3,4,5-kl]acridine-2-(6H) propanamine, 1-amino-9-ethyl-5-fluoro-2,3- dihydro-9-hydroxy-4-methyl-1H,12H-benzo[de]pyrano[3',4':b,7]-indolizino[1,2b]quinoline ,13(9H,15H)dione, lurtotecan, 7-[2-(N-isopropylamino)ethyl]-(20S)camptothecin, BNP1350, BNPI1100, BN80915, BN80942, etoposide phosphate, teniposide, sobuzoxane, 2' dimethylamino-2'-deoxy-etoposide, GL331, N-[2-(dimethylamino)ethyl]-9-hydroxy-5,6 dimethyl-6H-pyrido[4,3-b]carbazole-1-carboxamide, asulacrine, (5a, 5aB, 8aa,9b)-9-[2-[N-[2 (dimethylamino)ethyl]-N-methylamino]ethyl]-5-[4-hydroOxy-3,5-dimethoxyphenyl] ,5a,6,8,8a,9-hexohydrofuro(3',4':6,7)naphtho(2,3-d)-1,3-dioxol-6-one, 2,3-(methylenedioxy)-5 methyl-7-hydroxy-8-methoxybenzo[c]-phenanthridinium, 6,9-bis[(2 aminoethyl)amino]benzo[g]isoguinoline-5,10-dione, 5-(3-aminopropylamino)-7,10-dihydroxy-2 (2-hydroxyethylaminomethyl)-6H-pyrazolo[4,5,1-de]acridin-6-one, N-[1
[2(diethylamino)ethylamino]-7-methoxy-9-oxo-9H-thioxanthen-4-ylmethyl]formamide, N-(2 (dimethylamino)ethyl)acridine-4-carboxamide, 6-[[2-(dimethylamino)ethyl]amino]-3-hydroxy 7H-indeno[2,1-c] quinolin-7-one, and dimesna.
[0252] Examples of inhibitors of mitotic kinesins, and in particular the human mitotic kinesin KSP, that can be used in combination with the compounds of the invention include, but are not limited to, inhibitors described in PCT Publications WO 01/30768, WO 01/98278, WO 03/050,064, WO 03/050,122, WO 03/049,527, WO 03/049,679, WO 03/049,678, W004/039774, W003/079973, W003/099211, W003/105855, W003/106417, W004/037171, W004/058148, W004/058700, W004/126699, W005/018638, W005/019206, W005/019205, W005/018547, W005/017190, US2005/0176776. In an embodiment inhibitors of mitotic kinesins include, but are not limited to inhibitors of KSP, inhibitors of MKLP1, inhibitors of CENP-E, inhibitors of MCAK, inhibitors of Kifl4, inhibitors of Mphosphl and inhibitors of Rab6-KIFL.
[0253] Examples of "histone deacetylase inhibitors" that can be used in combination with the compounds of the invention include, but are not limited to, TSA, oxamflatin, PXD101, MG98, valproic acid and scriptaid. Further reference to other histone deacetylase inhibitors may be found in the following manuscript; Miller, T.A. et al. J. Med. Chem. 46(24):5097-5116 (2003).
[0254] Inhibitors of kinases involved in mitotic progression that can be used in combination with the compounds of the invention include, but are not limited to, inhibitors of aurora kinase, inhibitors of Polo-like kinases (PLK) (in particular inhibitors of PLK-1), inhibitors of bub-1 and inhibitors of bub-RI.
[0255] Antiproliferative agents that can be used in combination with the compounds of the invention include, but are not limited to, antisense RNA and DNA oligonucleotides such as G3139, ODN698, RVASKRAS, GEM231, and INX3001, and antimetabolites such as enocitabine, carmofur, tegafur, pentostatin, doxifluridine, trimetrexate, fludarabine, capecitabine, galocitabine, cytarabine ocfosfate, fosteabine sodium hydrate, raltitrexed, paltitrexid, emitefur, tiazofurin, decitabine, nolatrexed, pemetrexed, nelzarabine, 2'-deoxy-2'-methylidenecytidine, 2' fluoromethylene-2'-deoxycytidine, N-[5-(2,3-dihydro-benzofuryl)sulfonyl]-N'-(3,4 dichlorophenyl)urea, N6-[4-deoxy-4-[N2-[2(E),4(E)-tetradecadienoyl]glycylamino]-L-glycero B-L-manno-heptopyranosyl]adenine, aplidine, ecteinascidin, troxacitabine, 4-[2-amino-4-oxo 4,6,7,8-tetrahydro-3H-pyrimidino[5,4-b][1,4]thiazin-6-yl-(S)-ethyl]-2,5-thienoyl-L-glutamic acid, aminopterin, 5-flurouracil, alanosine, 11-acetyl-8-(carbamoyloxymethyl)-4-formyl-6 methoxy-14-oxa-1,11-diazatetracyclo(7.4.1.0.0)-tetradeca-2,4,6-trien-9-y acetic acid ester, swainsonine, lometrexol, dexrazoxane, methioninase, 2'-cyano-2'-deoxy-N4-palmitoyl-1-B-D arabino furanosyl cytosine and 3-aminopyridine-2-carboxaldehyde thiosemicarbazone.
[0256] Examples of monoclonal antibody targeted therapeutic agents that can be used in combination with the compounds of the invention include those therapeutic agents which have cytotoxic agents or radioisotopes attached to a cancer cell specific or target cell specific monoclonal antibody, such as, for example, Bexxar.
[0257] Examples of HMG-CoA reductase inhibitors that may be used that can be used in combination with the compounds of the invention include, but are not limited to, lovastatin
(MEVACOR; see U.S. Pat. Nos. 4,231,938, 4,294,926 and 4,319,039), simvastatin (ZOCOR®;
see U.S. Pat. Nos. 4,444,784, 4,820,850 and 4,916,239), pravastatin (PRAVACHOL®; see U.S.
Pat. Nos. 4,346,227, 4,537,859, 4,410,629, 5,030,447 and 5,180,589), fluvastatin (LESCOL®; see U.S. Pat. Nos. 5,354,772, 4,911,165, 4,929,437, 5,189,164, 5,118,853, 5,290,946 and
,356,896) and atorvastatin (LIPITOR; see U.S. Pat. Nos. 5,273,995, 4,681,893, 5,489,691 and ,342,952). The structural formulas of these and additional HMG-CoA reductase inhibitors that may be used in the instant methods are described at page 87 of M. Yalpani, "Cholesterol
Lowering Drugs", Chemistry & Industry, pp. 85-89 (5 February 1996) and U.S. Patent Nos. 4,782,084 and 4,885,314.
[0258] Examples of prenyl-protein transferase inhibitors that can be used in combination with the compounds of the invention include, but are not limited to, can be found in the following publications and patents: WO 96/30343, WO 97/18813, WO 97/21701, WO 97/23478, WO 97/38665, WO 98/28980, WO 98/29119, WO 95/32987, U.S. Pat. No. 5,420,245, U.S. Pat. No. 5,523,430, U.S. Pat. No. 5,532,359, U.S. Pat. No. 5,510,510, U.S. Pat. No. 5,589,485, U.S. Pat. No. 5,602,098, European Patent Publ. 0 618 221, European Patent Publ. 0 675 112, European Patent Publ. 0 604 181, European Patent Publ. 0 696 593, WO 94/19357, WO /08542, WO 95/11917, WO 95/12612, WO 95/12572, WO 95/10514, U.S. Pat. No. 5,661,152, WO 95/10515, WO 95/10516, WO 95/24612, WO 95/34535, WO 95/25086, WO 96/05529, WO 96/06138, WO 96/06193, WO 96/16443, WO 96/21701, WO 96/21456, WO 96/22278, WO 96/24611, WO 96/24612, WO 96/05168, WO 96/05169, WO 96/00736, U.S. Pat. No. 5,571,792, WO 96/17861, WO 96/33159, WO 96/34850, WO 96/34851, WO 96/30017, WO 96/30018, WO 96/30362, WO 96/30363, WO 96/31111, WO 96/31477, WO 96/31478, WO 96/31501, WO 97/00252, WO 97/03047, WO 97/03050, WO 97/04785, WO 97/02920, WO 97/17070, WO 97/23478, WO 97/26246, WO 97/30053, WO 97/44350, WO 98/02436, and U.S. Pat. No. ,532,359. For an example of the role of a prenyl-protein transferase inhibitor on angiogenesis see EuropeanJ. of Cancer, Vol. 35, No. 9, pp. 1 3 9 4 - 1 4 0 1 (1999).
[0259] Examples of angiogenesis inhibitors that can be used in combination with the compounds of the invention include, but are not limited to, tyrosine kinase inhibitors, such as inhibitors of the tyrosine kinase receptors Flt-1 (VEGFR1) and Flk-1/KDR (VEGFR2), inhibitors of epidermal-derived, fibroblast-derived, or platelet derived growth factors, MMP (matrix metalloprotease) inhibitors, integrin blockers, interferon-c, interleukin-12, pentosan polysulfate, cyclooxygenase inhibitors, including nonsteroidal anti-inflammatories (NSAIDs) like aspirin and ibuprofen as well as selective cyclooxy-genase-2 inhibitors like celecoxib and rofecoxib (PNAS, Vol. 89, p. 7384 (1992); JNCI, Vol. 69, p. 475 (1982); Arch. Opthalmol., Vol. 108, p.573 (1990); Anat. Rec., Vol. 238, p. 68 (1994); FEBS Letters, Vol. 372, p. 83 (1995); Clin, Orthop. Vol. 313, p. 76 (1995); J. Mol. Endocrinol., Vol. 16, p. 1 0 7 (1996); Jpn. J. Pharmacol., Vol. 75, p. 105 (1997); Cancer Res., Vol. 57, p. 1625 (1997); Cell, Vol. 93, p. 705 (1998); Intl. J. Mol. Med.,
Vol. 2, p. 715 (1998); J. Biol. Chem., Vol. 274, p. 9116 (1999)), steroidal anti-inflammatories (such as corticosteroids, mineralocorticoids, dexamethasone, prednisone, prednisolone, methylpred, betamethasone), carboxyamidotriazole, combretastatin A-4, squalamine, 6-0 chloroacetyl-carbonyl)-fumagillol, thalidomide, angiostatin, troponin-1, angiotensin II antagonists (see Fernandez et al., J. Lab. Clin. Med. 105:141-145 (1985)), and antibodies to VEGF (see, Nature Biotechnology, Vol. 17, pp. 9 6 3 - 9 6 8 (October 1999); Kim et al., Nature, 362, 841-844 (1993); WO 00/44777; and WO 00/61186).
[0260] Other therapeutic agents that modulate or inhibit angiogenesis may also be used in combination with the compounds of the instant invention and include agents that modulate or inhibit the coagulation and fibrinolysis systems (see review in Clin. Chem. La. Med. 38:679-692 (2000)). Examples of such agents that modulate or inhibit the coagulation andfibrinolysis pathways that can be used in combination with the compounds of the invention include, but are not limited to, heparin (see Thromb. Haemost. 80:10-23 (1998)), low molecular weight heparins and carboxypeptidase U inhibitors (also known as inhibitors of active thrombin activatable fibrinolysis inhibitor [TAFIa]) (see Thrombosis Res. 101:329-354 (2001)). TAFIa inhibitors have been described in PCT Publication WO 03/013,526 and U.S. Ser. No. 60/349,925 (filed January 18, 2002).
[0261] Agents that interfere with cell cycle checkpoints that can be used in combination with the compounds of the invention include, but are not limited to, inhibitors of ATR, ATM, the Chkl and Chk2 kinases and cdk and cdc kinase inhibitors and are specifically exemplified by 7 hydroxystaurosporin, flavopiridol, CYC202 (Cyclacel) and BMS-387032.
[0262] Agents that interfere with receptor tyrosine kinases (RTKs) that can be used in combination with the compounds of the invention include, but are not limited to, inhibitors of c Kit, Eph, PDGF, Flt3 and HBV. Further agents include inhibitors of RTKs as described by Bume-Jensen and Hunter, Nature, 411:355-365, 2001.
[0263] Inhibitors of cell proliferation and survival signaling pathway that can be used in combination with the compounds of the invention include, but are not limited to, inhibitors of EGFR (for example gefitinib and erlotinib), inhibitors of ERB-2 (for example trastuzumab), inhibitors of IGFR, inhibitors of cytokine receptors, inhibitors of HBV, inhibitors of P13K (for example LY294002), serine/threonine kinases (including but not limited to inhibitors of Akt such as described in WO 02/083064, WO 02/083139, WO 02/083140, US 2004-0116432, WO 02/083138, US 2004-0102360, WO 03/086404, WO 03/086279, WO 03/086394, WO 03/084473, WO 03/086403, WO 2004/041162, WO 2004/096131, WO 2004/096129, WO 2004/096135, WO 2004/096130, WO 2005/100356, WO 2005/100344), inhibitors of Raf kinase (for example BAY-43-9006), inhibitors of MEK (for example CI-1040 and PD-098059) and inhibitors of mTOR (for example Wyeth CCI-779). Such agents include small molecule inhibitor compounds and antibody antagonists.
[0264] Apoptosis inducing agents that can be used in combination with the compounds of the invention include, but are not limited to, activators of TNF receptor family members (including the TRAIL receptors).
[0265] NSAIDs that are selective COX-2 inhibitors that can be used in combination with the compounds of the invention include, but are not limited to, those NSAIDs disclosed in U.S. Pat. ,474,995, U.S. Pat. 5,861,419, U.S. Pat. 6,001,843, U.S. Pat. 6,020,343, U.S. Pat. 5,409,944, U.S. Pat. 5,436,265, U.S. Pat. 5,536,752, U.S. Pat. 5,550,142, U.S. Pat. 5,604,260, U.S. ,698,584, U.S. Pat. 5,710,140, WO 94/15932, U.S. Pat. 5,344,991, U.S. Pat. 5,134,142, U.S. Pat. 5,380,738, U.S. Pat. 5,393,790, U.S. Pat. 5,466,823, U.S. Pat. 5,633,272, and U.S. Pat. ,932,598, all of which are hereby incorporated by reference.
[0266] Inhibitors of COX-2 that are particularly useful in combination with the compounds of the invention include: 3-phenyl-4-(4-(methylsulfonyl)phenyl)-2-(5H)-furanone; and 5-chloro 3-(4-methylsulfonyl)-phenyl-2-(2-methyl-5-pyridinyl)pyridine; or a pharmaceutically acceptable salt thereof.
[0267] Compounds that have been described as specific inhibitors of COX-2 and are therefore useful in the present invention include, but are not limited to: parecoxib, CELEBREX* and BEXTRA* or a pharmaceutically acceptable salt thereof.
[0268] Angiogenesis inhibitors that can be used in combination with the compounds of the invention include, but are not limited to, endostatin, ukrain, ranpirnase, IM862, 5-methoxy-4-[2 methyl-3-(3-methyl-2-butenyl)oxiranyl]-1-oxaspiro[2,5]oct-6-yl(chloroacetyl)carbamate, acetyldinanaline, 5-amino-I-[[3,5-dichloro-4-(4-chlorobenzoyl)-phenyl]methyl]-1H-1,2,3 triazole-4-carboxamide,CM101, squalamine, combretastatin, RP14610, NX31838, sulfated mannopentaose phosphate, 7,7-(carbonyl-bis[imino-N-methyl-4,2-pyrrolocarbonylimino[N methyl-4,2-pyrrole]-carbonylimino]-bis-(1,3-naphthalene disulfonate), and 3-[(2,4 dimethylpyrrol-5-yl)methylene]-2-indolinone (SU5416).
[0269] Tyrosine kinase inhibitors that can be used in combination with the compounds of the invention include, but are not limited to, N-(trifluoromethylphenyl)-5-methylisoxazol-4 carboxamide, 3-[(2,4-dimethylpyrrol-5-yl)methylidenyl)indolin-2-one, 17-(allylamino)-17 demethoxygeldanamycin, 4-(3-chloro-4-fluorophenylamino)-7-methoxy-6-[3-(4 morpholinyl)propoxyl]quinazoline, N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4 quinazolinamine, BIBX1382, 2,3,9,10,11,12-hexahydro-10-(hydroxymethyl)-10-hydroxy-9 methyl-9,12-epoxy-IH-diindolo[1,2,3-fg:3',2',1'-kl]pyrrolo[3,4-i][1,6]benzodiazocin-1-one, SH268, genistein, imatinib (STI571), CEP2563, 4-(3-chlorophenylamino)-5,6-dimethyl-7H pyrrolo[2,3-d]pyrimidinemethane sulfonate, 4-(3-bromo-4-hydroxyphenyl)amino-6,7 dimethoxyquinazoline, 4-(4'-hydroxyphenyl)amino-6,7-dimethoxyquinazoline, SU6668, ST1571A,N-4-chlorophenyl-4-(4-pyridylmethyl)-1-phthalazinamine,andEMD121974.
[0270] Combinations with compounds other than anti-cancer compounds are also encompassed in the instant compositions and methods. For example, combinations of the instantly claimed compounds with PPAR-y (i.e., PPAR-gamma) agonists and PPAR-6 (i.e.,
PPAR-delta) agonists are useful in the treatment of certain malignancies. PPAR-y and PPAR-6
are the nuclear peroxisome proliferator-activated receptors y and 6. The expression of PPAR-y on endothelial cells and its involvement in angiogenesis has been reported in the literature (see J. Cardiovasc. Pharmacol. 31:909-913 (1998); J. Biol. Chem. 274:9116-9121 (1999); Invest. Ophthalmol Vis. Sci. 41:2309-2317 (2000)). More recently, PPAR-y agonists have been shown to inhibit the angiogenic response to VEGF in vitro; both troglitazone and rosiglitazone maleate inhibit the development of retinal neovascularization in mice. (Arch. Ophthamol. 119:709-717 (2001)). Examples of PPAR-y agonists and PPAR- 7/c agonists that can be used in combination with the compounds of the invention include, but are not limited to, thiazolidinediones (such as DRF2725, CS-011, troglitazone, rosiglitazone, and pioglitazone), fenofibrate, gemfibrozil, clofibrate, GW2570, SB219994, AR-H039242, JTT-501, MCC-555, GW2331, GW409544,
NN2344, KRP297, NP0110, DRF4158, NN622, G1262570, PNU182716, DRF552926, 2-[(5,7 dipropyl-3-trifluoromethyl-1,2-benzisoxazol-6-yl)oxy]-2-methylpropionic acid (disclosed in USSN 09/782,856), and 2(R)-7-(3-(2-chloro-4-(4-fluorophenoxy) phenoxy)propoxy)-2 ethylchromane-2-carboxylic acid (disclosed in USSN 60/235,708 and 60/244,697).
[0271] Another embodiment of the instant invention is the use of the presently disclosed compounds in combination with gene therapy for the treatment of cancer. For an overview of genetic strategies to treating cancer see Hall et al. (Am J Hum Genet 61:785-789 (1997)) and Kufe et al. (Cancer Medicine, 5th Ed, pp 876-889, BC Decker, Hamilton, 2000). Gene therapy can be used to deliver any tumor suppressing gene. Examples of such genes include, but are not limited to, p53, which can be delivered via recombinant virus-mediated gene transfer (see U.S. Pat. No. 6,069,134, for example), a uPA/uPAR antagonist ("Adenovirus-Mediated Delivery of a uPA/uPAR Antagonist Suppresses Angiogenesis-Dependent Tumor Growth and Dissemination in Mice," Gene Therapy, August 5(8):1105-13 (1998)), and interferon gamma (J Immunol 164:217-222 (2000)).
[0272] The compounds of the instant invention may also be administered in combination with an inhibitor of inherent multidrug resistance (MDR), in particular MDR associated with high levels of expression of transporter proteins. Such MDR inhibitors include inhibitors of p glycoprotein (P-gp), such as LY335979, XR9576, OC144-093, R101922, VX853 and PSC833 (valspodar).
[0273] A compound of the present invention may be employed in conjunction with anti emetic agents to treat nausea or emesis, including acute, delayed, late-phase, and anticipatory emesis, which may result from the use of a compound of the present invention, alone or with radiation therapy. For the prevention or treatment of emesis, a compound of the present invention may be used in conjunction with other anti-emetic agents, especially neurokinin-1 receptor antagonists, 5HT3 receptor antagonists, such as ondansetron, granisetron, tropisetron, and zatisetron, GABAB receptor agonists, such as baclofen, a corticosteroid such as Decadron (dexamethasone), Kenalog, Aristocort, Nasalide, Preferid, Benecorten or others such as disclosed in U.S. Patent Nos. 2,789,118, 2,990,401, 3,048,581, 3,126,375, 3,929,768, 3,996,359, 3,928,326 and 3,749,712, an antidopaminergic, such as the phenothiazines (for example prochlorperazine, fluphenazine, thioridazine and mesoridazine), metoclopramide or dronabinol. In an embodiment, an anti-emesis agent selected from a neurokinin-1 receptor antagonist, a 5HT3 receptor antagonist and a corticosteroid is administered as an adjuvant for the treatment or prevention of emesis that may result upon administration of the instant compounds.
[0274] Neurokinin-1 receptor antagonists of use in conjunction with the compounds of the present invention are fully described, for example, in U.S. Pat. Nos. 5,162,339, 5,232,929, ,242,930, 5,373,003, 5,387,595, 5,459,270, 5,494,926, 5,496,833, 5,637,699, 5,719,147; European Patent Publication Nos. EP 0 360 390, 0 394 989, 0 428 434, 0 429 366, 0 430 771, 0 436 334, 0 443 132, 0 482 539, 0 498 069, 0 499 313, 0 512 901, 0 512 902, 0514 273, 0 514 274,0514275,0514276,0515 681,0517589,0520555,0522 808,0528 495,0532456,0 533 280, 0 536 817, 0 545 478, 0 558 156, 0 577 394, 0 585 913,0 590 152, 0 599538, 0 610 793,0634402,0686629,0693489,0694535,0699655,0699674,0707006,0708 101,0 709 375, 0 709 376, 0 714 891, 0 723 959, 0 733 632 and 0 776 893; PCT International Patent Publication Nos. WO 90/05525, 90/05729, 91/09844, 91/18899, 92/01688, 92/06079, 92/12151, 92/15585, 92/17449, 92/20661, 92/20676, 92/21677, 92/22569, 93/00330, 93/00331, 93/01159, 93/01165, 93/01169, 93/01170, 93/06099, 93/09116, 93/10073, 93/14084, 93/14113, 93/18023, 93/19064, 93/21155, 93/21181, 93/23380, 93/24465, 94/00440, 94/01402, 94/02461, 94/02595, 94/03429, 94/03445, 94/04494, 94/04496, 94/05625, 94/07843, 94/08997, 94/10165, 94/10167, 94/10168, 94/10170, 94/11368, 94/13639, 94/13663, 94/14767, 94/15903, 94/19320, 94/19323, 94/20500, 94/26735, 94/26740, 94/29309, 95/02595, 95/04040, 95/04042, 95/06645, 95/07886, /07908, 95/08549, 95/11880, 95/14017, 95/15311, 95/16679, 95/17382, 95/18124, 95/18129, /19344, 95/20575, 95/21819, 95/22525, 95/23798, 95/26338, 95/28418, 95/30674, 95/30687, /33744, 96/05181, 96/05193, 96/05203, 96/06094, 96/07649, 96/10562, 96/16939, 96/18643, 96/20197, 96/21661, 96/29304, 96/29317, 96/29326, 96/29328, 96/31214, 96/32385, 96/37489, 97/01553, 97/01554, 97/03066, 97/08144, 97/14671, 97/17362, 97/18206, 97/19084, 97/19942 and 97/21702; and in British Patent Publication Nos. 2 266 529, 2 268 931, 2 269 170, 2 269 590, 2 271 774, 2 292 144, 2 293 168, 2 293 169, and 2 302 689. The preparation of such compounds is fully described in the aforementioned patents and publications, which are incorporated herein by reference.
[0275] In an embodiment, the neurokinin-1 receptor antagonist for use in conjunction with the compounds of the present invention is selected from: 2-(R)-(1-(R)-(3,5-bis(trifluoromethyl) phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(5-oxo-1H,4H-1,2,4-triazolo)methyl)morpholine, or a pharmaceutically acceptable salt thereof, which is described in U.S. Pat. No. 5,719,147.
[0276] A compound of the instant invention may also be administered with an agent useful in the treatment of anemia. Such an anemia treatment agent is, for example, a continuous eythropoiesis receptor activator (such as epoetin alfa).
[0277] A compound of the instant invention may also be administered with an agent useful in the treatment of neutropenia. Such a neutropenia treatment agent is, for example, a hematopoietic growth factor which regulates the production and function of neutrophils such as a human granulocyte colony stimulating factor, (G-CSF). Examples of a G-CSF include filgrastim and PEG-filgrastim.
[0278] A compound of the instant invention may also be administered with an immunologic enhancing drug, such as levamisole, isoprinosine and Zadaxin.
[0279] A compound of the instant invention may also be useful for treating or preventing liver disease or cancer in combination with other siNA therapeutics.
[0280] The compounds of the instant invention may also be administered in combination with 7-secretase inhibitors and/or inhibitors of NOTCH signaling. Such inhibitors include compounds described in WO 01/90084, WO 02/30912, WO 01/70677, WO 03/013506, WO 02/36555, WO 03/093252, WO 03/093264, WO 03/093251, WO 03/093253, WO 2004/039800, WO 2004/039370, WO 2005/030731, WO 2005/014553, USSN 10/957,251, WO 2004/089911, WO 02/081435, WO 02/081433, WO 03/018543, WO 2004/031137, WO 2004/031139, WO 2004/031138, WO 2004/101538, WO 2004/101539 and WO 02/47671 (including LY-450139).
[0281] A compound of the instant invention may also be useful for treating or preventing cancer in combination with PARP inhibitors.
[0282] A compound of the instant invention may also be useful for treating cancer in combination with the following therapeutic agents: abarelix (Plenaxis depot); aldesleukin
(Prokine); Aldesleukin (Proleukin®); Alemtuzumabb (Campath®); alitretinoin (Panretin®); allopurinol (Zyloprim®); altretamine (Hexalen®); amifostine (Ethyol®); anastrozole (Arimidex); arsenic trioxide (Trisenox®); asparaginase (Elspar®); azacitidine (Vidaza); bendamustine hydrochloride (Treanda®); bevacuzimab (Avastin®); bexarotene capsules (Targretin@); bexarotene gel (Targretin®); bleomycin (Blenoxane®); bortezomib (Velcade®); brefeldin A; busulfan intravenous (Busulfex®); busulfan oral (Myleran®); calusterone (Methosarb®); capecitabine (Xeloda®); carboplatin (Paraplatin®); carmustine (BCNU®, BiCNU®); carmustine (Gliadel®); carmustine with Polifeprosan 20 Implant (Gliadel Wafer®); celecoxib (Celebrex®); cetuximab (Erbitux®); chlorambucil (Leukeran®); cisplatin (Platinol®); cladribine (Leustatin®, 2-CdA®); clofarabine (Clolar®); cyclophosphamide (Cytoxan®, Neosar®); cyclophosphamide (Cytoxan Injection®); cyclophosphamide (Cytoxan Tablet); cytarabine (Cytosar-U®); cytarabine liposomal (DepoCyt®); dacarbazine (DTIC-Dome®); dactinomycin, actinomycin D (Cosmegen®); dalteparin sodium injection (Fragmin®); Darbepoetin alfa (Aranesp®); dasatinib (Sprycel®); daunorubicin liposomal (DanuoXome®); daunorubicin, daunomycin (Daunorubicin®); daunorubicin, daunomycin (Cerubidine®); degarelix (Firmagon); Denileukin diftitox (Ontak®); dexrazoxane (Zinecard®); dexrazoxane hydrochloride (Totect®); didemnin B; 17-DMAG; docetaxel (Taxotere®); doxorubicin (Adriamycin PFS); doxorubicin (Adriamycin, Rubex®); doxorubicin (Adriamycin PFS Injections); doxorubicin liposomal (Doxil®); dromostanolone propionate (Dromostanolone ); dromostanolone propionate (Masterone Injection); eculizumab injection (Soliris®); Elliott's B Solution (Elliott's B Solution®); eltrombopag (Promacta®); epirubicin (Ellence®); Epoetin alfa (epogen®); erlotinib (Tarceva®); estramustine (Emcyt@); ethinyl estradiol; etoposide phosphate (Etopophos®); etoposide, VP-16 (Vepesid®); everolimus tablets (Afinitor®); exemestane (Aromasin®); ferumoxytol (Feraheme Injection®); Filgrastim (Neupogen®); floxuridine (intraarterial) (FUDR); fludarabine (Fludara); fluorouracil, 5-FU (Adrucil®); fulvestrant (Faslodex®); gefitinib (Iressa®); geldanamycin; gemcitabine (Gemzar@); gemtuzumab ozogamicin (Mylotarg); goserelin acetate (Zoladex Implant®); goserelin acetate (Zoladex); histrelin acetate (Histrelin implants); hydroxyurea (Hydrea®); Ibritumomab Tiuxetan (Zevalin®); idarubicin (Idamycin®); ifosfamide (IFEX®); imatinib mesylate (Gleevec®); interferon alfa 2a (Roferon A®); Interferon alfa-2b (Intron A®); iobenguane I 123 injection (AdreView); irinotecan (Camptosar®); ixabepilone (Ixempra®); lapatinib tablets (Tykerb); lenalidomide (Revlimid); letrozole (Femara®); leucovorin (Wellcovorin®, Leucovorin); Leuprolide Acetate (Eligard®); levamisole (Ergamisol®); lomustine, CCNU (CeeBU®); meclorethamine, nitrogen mustard (Mustargen); megestrol acetate (Megace®); melphalan, L PAM (Alkeran®); mercaptopurine, 6-MP (Purinethol@); mesna (Mesnex®); mesna (Mesnex tabs®); methotrexate (Methotrexate®); methoxsalen (Uvadex®); 8-methoxypsoralen; mitomycin
C (Mutamycin®); mitotane (Lysodren®); mitoxantrone (Novantrone®); mitramycin; nandrolone phenpropionate (Durabolin-50O); nelarabine (Arranon®); nilotinib (Tasigna®); Nofetumomab (Verluma®); ofatumumab (Arzerra®); Oprelvekin (Neumega®); oxaliplatin (Eloxatin®); paclitaxel (Paxene); paclitaxel (Taxol®); paclitaxel protein-bound particles (Abraxane®); palifermin (Kepivance®); pamidronate (Aredia®); panitumumab (Vectibix®); pazopanib tablets (Votrienttm@); pegademase (Adagen (Pegademase Bovine)®); pegaspargase (Oncaspar); Pegfilgrastim (Neulasta®); pemetrexed disodium (Alimta®); pentostatin (Nipent®); pipobroman (Vercyte); plerixafor (Mozobil®); plicamycin, mithramycin (Mithracin®); porfimer sodium (Photofrin®); pralatrexate injection (Folotyn®); procarbazine (Matulane®); quinacrine (Atabrine®); rapamycin; Rasburicase (Elitek®); raloxifene hydrochloride (Evista®); Rituximab (Rituxan®); romidepsin (Istodax®); romiplostim (Nplate@); sargramostim (Leukine®); Sargramostim (Prokine); sorafenib (Nexavar®); streptozocin (Zanosar®); sunitinib maleate (Sutent®); talc (Sclerosol@); tamoxifen (Nolvadex®); temozolomide (Temodar®); temsirolimus (Torisel®); teniposide, VM-26 (Vumon®); testolactone (Teslac®); thioguanine, 6-TG (Thioguanine®); thiopurine; thiotepa (Thioplex®); topotecan (Hycamtin®); toremifene (Fareston); Tositumomab (Bexxar®); Tositumomab/I-131 tositumomab (Bexxar®); trans retinoic acid; Trastuzumab (Herceptin®); tretinoin, ATRA (Vesanoid); triethylenemelamine; Uracil Mustard (Uracil Mustard Capsules®); valrubicin (Valstar®); vinblastine (Velban®); vincristine (Oncovin); vinorelbine (Navelbine); vorinostat (Zolinza®); wortmannin; and zoledronate (Zometa®).
[0283] The combinations referred to above can conveniently be presented for use in the form of a pharmaceutical formulation and thus pharmaceutical compositions comprising a combination as defined above together with a pharmaceutically acceptable diluent or carrier represent a further aspect of the invention.
[0284] The individual compounds of such combinations can be administered either sequentially or simultaneously in separate or combined pharmaceutical formulations. In one embodiment, the individual compounds will be administered simultaneously in a combined pharmaceutical formulation.
[0285] Thus, the described molecules could be used in combination with one or more known compounds, treatments, or procedures to prevent or treat diseases, disorders, conditions, and traits described herein in a subject or organism as are known in the art, such as other HBV inhibitors.
3. Therapeutic Applications
[0286] The present body of knowledge in HBV research indicates the need for methods that can regulate HBV expression for therapeutic use.
[0287] Thus, one aspect of the invention comprises a method of treating a subject including, but not limited to, a human suffering HBV infection or a condition which is mediated by the action of HBV gene expression, which method comprises administering to said subject an effective amount of a double-stranded siNA molecule of the invention. In one embodiment of this aspect, the siNA molecules comprises at least a 15 nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 452, SEQ ID NO: 2, SEQ ID NO: 453, SEQ ID NO: 3, SEQ ID NO: 454, SEQ ID NO: 4, or SEQ ID NO:455; or formula (A).
[0288] In some embodiments of this aspect, the condition is cancer. Thus, in certain embodiments the molecules and compositions of the instant invention are useful in a method for treating cancer, and in particular, in treating hepatocellular carcinoma (HCC).
[0289] In some embodiments, the condition is liver disease. Thus, in certain embodiments the molecules and compositions of the instant invention are useful in a method for treating liver disease, and, in particular, in treating cirrhosis of the liver.
[0290] In certain embodiments, the administration of the siNA molecule is via local administration or systemic administration. In other embodiments, the invention features contacting the subject or organism with an siNA molecule of the invention via local administration to relevant tissues or cells, such as lung cells and tissues, such as via pulmonary delivery. In yet other embodiments, the invention features contacting the subject or organism with an siNA molecule of the invention via systemic administration (such as via intravenous or subcutaneous administration of siNA) to relevant tissues or cells, such as liver or cancerous tissues or cells in a subject or organism.
[0291] siNA molecules of the invention are also used as reagents in ex vivo applications. For example, siNA reagents are introduced into tissue or cells that are transplanted into a subject for therapeutic effect. The cells and/or tissue can be derived from an organism or subject that later receives the explant, or can be derived from another organism or subject prior to transplantation.
The siNA molecules can be used to modulate the expression of one or more genes in the cells or tissue, such that the cells or tissue obtain a desired phenotype or are able to perform a function when transplanted in vivo. In one embodiment, certain HBV target cells from a patient are extracted. These extracted cells are contacted with HBV siNAs targeting a specific nucleotide sequence within the cells under conditions suitable for uptake of the siNAs by these cells (e.g., using delivery reagents such as cationic lipids, liposomes and the like or using techniques such as electroporation to facilitate the delivery of siNAs into cells). The cells are then reintroduced back into the same patient or other patients.
[0292] For therapeutic applications, a pharmaceutically effective dose of the siNA molecules or pharmaceutical compositions of the invention is administered to the subject. A pharmaceutically effective dose is that dose required to prevent, inhibit the occurrence, or treat (alleviate a symptom to some extent, preferably all of the symptoms) a disease state. One skilled in the art can readily determine a therapeutically effective dose of the siNA of the invention to be administered to a given subject, by taking into account factors, such as the size and weight of the subject, the extent of the disease progression or penetration, the age, health, and sex of the subject, the route of administration, and whether the administration is regional or systemic. Generally, an amount between 0.1 pg/kg and 100 mg/kg body weight/day of active ingredients is administered dependent upon potency of the negatively charged polymer. Optimal dosing schedules can be calculated from measurements of drug accumulation in the body of the patient. The siNA molecules of the invention can be administered in a single dose or in multiple doses.
[0293] siNA molecules of the instant invention can be administered once monthly, once weekly, once daily (QD), or divided into multiple monthly, weekly, or daily doses, such as, for example, but not limitation, twice daily (BID), three times daily (TID), once every two weeks. Persons of ordinary skill in the art can easily estimate repetition rates for dosing based on measured residence times and concentrations of the drug in bodily fluids or tissues.
[0294] In addition, the administration can be continuous, i.e., every day, or intermittently. For example, intermittent administration of a compound of the instant invention may be administration one to six days per week or it may mean administration in cycles (e.g. daily administration for two to eight consecutive weeks, then a rest period with no administration for up to one week) or it may mean administration on alternate days.
G. Administration
[0295] Compositions or formulations can be administered in a variety of ways. Non-limiting examples of administration methods of the invention include oral, buccal, sublingual, parenteral (i.e., intraarticularly, intravenously, intraperitoneally, subcutaneously, or intramuscularly), local rectal administration or other local administration. In one embodiment, the composition of the invention can be administered by insufflation and inhalation. Administration can be accomplished via single or divided doses. In some embodiments, the pharmaceutical compositions are administered intravenously or intraperitoneally by a bolus injection (see, e.g., U.S. Pat. No. 5,286,634). Lipid nucleic acid particles can be administered by direct injection at the site of disease or by injection at a site distal from the site of disease (see, e.g., Culver, HUMAN GENE THERAPY, MaryAnn Liebert, Inc., Publishers, New York. pp. 70-71(1994)). In one embodiment, the siNA molecules of the invention and formulations or compositions thereof are administered to a cell, subject, or organism as is described herein and as is generally known in the art.
1. In Vivo Administration
[0296] In any of the methods of treatment of the invention, the siNA can be administered to the subject systemically as described herein or otherwise known in the art, either alone as a monotherapy or in combination with additional therapies described herein or as are known in the art. Systemic administration can include, for example, pulmonary (inhalation, nebulization etc.) intravenous, subcutaneous, intramuscular, catheterization, nasopharangeal, transdermal, or oral/gastrointestinal administration as is generally known in the art.
[0297] In any of the methods of treatment or prevention of the invention, the siNA can be administered to the subject locally or to local tissues as described herein or otherwise known in the art, either alone as a monotherapy or in combination with additional therapies as are known in the art. Local administration can include, for example, inhalation, nebulization, catheterization, implantation, direct injection, dermal/transdermal application, patches, stenting, ear/eye drops, or portal vein administration to relevant tissues, or any other local administration technique, method or procedure, as is generally known in the art.
[0298] In one embodiment, the siNA molecules of the invention and formulations or compositions thereof are administered to the liver as is generally known in the art (see for example Wen et al., 2004, World J Gastroenterol., 10, 244-9; Murao et al., 2002, Pharm Res., 19, 1808-14; Liu et al., 2003, gene Ther., 10, 180-7; Hong et al., 2003, JPharmPharmacol., 54, 51-8; Herrmann et al., 2004, Arch Virol., 149, 1611-7; and Matsuno et al., 2003, gene Ther., 10, 1559-66).
[0299] In one embodiment, the invention features the use of methods to deliver the siNA molecules of the instant invention to hematopoietic cells, including monocytes and lymphocytes. These methods are described in detail by Hartmann et al., 1998, J. Phamacol. Exp. Ther., 285(2), 920-928; Kronenwett et al., 1998, Blood, 91(3), 852-862; Filion and Phillips, 1997, Biochim. Biophys. Acta., 1329(2), 345-356; Ma and Wei, 1996, Leuk. Res., 20(11/12), 925-930; and Bongartz et al., 1994, Nucleic Acids Research, 22(22), 4681-8.
[0300] In one embodiment, the siNA molecules of the invention and formulations or compositions thereof are administered directly or topically (e.g., locally) to the dermis or follicles as is generally known in the art (see for example Brand, 2001, Curr. Opin. Mol. Ther., 3, 244-8; Regnier et al., 1998, J. Drug Target, 5, 275-89; Kanikkannan, 2002, BioDrugs, 16, 339-47; Wraight et al., 2001, Pharmacol. Ther., 90, 89-104; and Preat and Dujardin, 2001, STP PharmaSciences, 11, 57-68). In one embodiment, the siNA molecules of the invention and formulations or compositions thereof are administered directly or topically using a hydroalcoholic gel formulation comprising an alcohol (e.g., ethanol or isopropanol), water, and optionally including additional agents such isopropyl myristate and carbomer 980. In other embodiments, the siNA are formulated to be administered topically to the nasal cavity. Topical preparations can be administered by one or more applications per day to the affected area; over skin areas occlusive dressings can advantageously be used. Continuous or prolonged delivery can be achieved by an adhesive reservoir system.
[0301] In one embodiment, an siNA molecule of the invention is administered iontophoretically, for example to a particular organ or compartment (e.g., the eye, back of the eye, heart, liver, kidney, bladder, prostate, tumor, CNS etc.). Non-limiting examples of iontophoretic delivery are described in, for example, WO 03/043689 and WO 03/030989, which are incorporated by reference in their entireties herein.
[0302] In one embodiment, the siNA molecules of the invention and formulations or compositions thereof are administered to the lung as is described herein and as is generally known in the art. In another embodiment, the siNA molecules of the invention and formulations or compositions thereof are administered to lung tissues and cells as is described in U.S. Patent Publication Nos. 2006/0062758; 2006/0014289; and 2004/0077540.
2. Aerosols and Delivery Devices
a. AerosolFormulations
[0303] The compositions of the present invention, either alone or in combination with other suitable components, can be made into aerosol formulations (i.e., they can be "nebulized") to be administered via inhalation (e.g., intranasally or intratracheally) (see, Brigham et al., Am. J. Sci., 298:278 (1989)). Aerosol formulations can be placed into pressurized acceptable propellants, such as dichlorodifluoromethane, propane, nitrogen, and the like.
[0304] In one embodiment, the siNA molecules of the invention and formulations thereof are administered via pulmonary delivery, such as by inhalation of an aerosol or spray dried formulation administered by an inhalation device or nebulizer, providing rapid local uptake of the nucleic acid molecules into relevant pulmonary tissues. Solid particulate compositions containing respirable dry particles of micronized nucleic acid compositions can be prepared by grinding dried or lyophilized nucleic acid compositions, and then passing the micronized composition through, for example, a 400 mesh screen to break up or separate out large agglomerates. A solid particulate composition comprising the siNA compositions of the invention can optionally contain a dispersant which serves to facilitate the formation of an aerosol as well as other therapeutic compounds. A suitable dispersant is lactose, which can be blended with the nucleic acid compound in any suitable ratio, such as a 1 to 1 ratio by weight.
[0305] Spray compositions comprising siNA molecules or compositions of the invention can, for example, be formulated as aqueous solutions or suspensions or as aerosols delivered from pressurized packs, such as a metered dose inhaler, with the use of a suitable liquefied propellant. In one embodiment, aerosol compositions of the invention suitable for inhalation can be either a suspension or a solution and generally contain an siNA molecule comprising at least a 15 nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 452, SEQ ID NO: 2, SEQ ID NO: 453, SEQ ID NO: 3, SEQ ID NO: 454, SEQ ID NO: 4, or SEQ ID NO:455; or formula (A), and a suitable propellant such as a fluorocarbon or hydrogen-containing chlorofluorocarbon or mixtures thereof, particularly hydrofluoroalkanes, especially 1,1,1,2-tetrafluoroethane, 1,1,1,2,3,3,3 heptafluoro-n-propane or a mixture thereof. The aerosol composition can optionally contain additional formulation excipients well known in the art such as surfactants. Non-limiting examples include oleic acid, lecithin or an oligolactic acid or derivative such as those described in WO94/21229 and WO98/34596 and co-solvents for example ethanol. In one embodiment a pharmaceutical aerosol formulation of the invention comprising a compound of the invention and a fluorocarbon or hydrogen-containing chlorofluorocarbon or mixtures thereof as propellant, optionally in combination with a surfactant and/or a co-solvent.
[0306] The aerosol formulations of the invention can be buffered by the addition of suitable buffering agents.
[0307] Aerosol formulations can include optional additives including preservatives if the formulation is not prepared sterile. Non-limiting examples include, methyl hydroxybenzoate, anti-oxidants, flavorings, volatile oils, buffering agents and emulsifiers and other formulation surfactants. In one embodiment, fluorocarbon or perfluorocarbon carriers are used to reduce degradation and provide safer biocompatible non-liquid particulate suspension compositions of the invention (e.g., siNA and/or LNP formulations thereof). In another embodiment, a device comprising a nebulizer delivers a composition of the invention (e.g., siNA and/or LNP formulations thereof) comprising fluorochemicals that are bacteriostatic thereby decreasing the potential for microbial growth in compatible devices.
[0308] Capsules and cartridges comprising the composition of the invention for use in an inhaler or insufflator, of for example gelatine, can be formulated containing a powder mix for inhalation of a compound of the invention and a suitable powder base such as lactose or starch. In one embodiment, each capsule or cartridge contains an siNA molecule comprising at least a nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 452, SEQ ID NO: 2, SEQ ID NO: 453, SEQ ID NO: 3, SEQ ID NO: 454, SEQ ID NO: 4, or SEQ ID NO:455; or formula (A), and one or more excipients. In another embodiment, the compound of the invention can be presented without excipients such as lactose
[0309] The aerosol compositions of the present invention can be administered into the respiratory system as a formulation including particles of respirable size, e.g. particles of a size sufficiently small to pass through the nose, mouth and larynx upon inhalation and through the bronchi and alveoli of the lungs. In general, respirable particles range from about 0.5 to 10 microns in size. In one embodiment, the particulate range can be from 1 to 5 microns. In another embodiment, the particulate range can be from 2 to 3 microns. Particles of non respirable size which are included in the aerosol tend to deposit in the throat and be swallowed, and the quantity of non-respirable particles in the aerosol is thus minimized. For nasal administration, a particle size in the range of 10-500 um is preferred to ensure retention in the nasal cavity.
[0310] In some embodiments, an siNA composition of the invention is administered topically to the nose for example, for the treatment of rhinitis, via pressurized aerosol formulations, aqueous formulations administered to the nose by pressurized pump or by nebulization. Suitable formulations contain water as the diluent or carrier for this purpose. In certain embodiments, the aqueous formulations for administration of the composition of the invention to the lung or nose can be provided with conventional excipients such as buffering agents, tonicity modifying agents and the like.
b. Devices
[0311] The siNA molecules of the invention can be formulated and delivered as particles and/or aerosols as discussed above and dispensed from various aerosolization devices known by those of skill in the art.
[0312] Aerosols of liquid or non-liquid particles comprising an siNA molecule or formulation of the invention can be produced by any suitable means, such as with a device comprising a nebulizer (see for example U.S. Pat. 4,501,729) such as ultrasonic or air jet nebulizers.
[0313] Solid particle aerosols comprising an siNA molecule or formulation of the invention and surfactant can be produced with any solid particulate aerosol generator. One type of solid particle aerosol generator used with the siNA molecules of the invention is an insufflator. A second type of illustrative aerosol generator comprises a metered dose inhaler ("MDI"). MDIs containing siNA molecules or formulations taught herein can be prepared by methods of the art (for example, see Byron, above and W096/32099).
[0314] The siNA molecules can also be formulated as a fluid formulation for delivery from a fluid dispenser, such as those described and illustrated in W005/044354.
[0315] In certain embodiments of the invention, nebulizer devices are used in applications for conscious, spontaneously breathing subjects, and for controlled ventilated subjects of all ages. The nebulizer devices can be used for targeted topical and systemic drug delivery to the lung. In one embodiment, a device comprising a nebulizer is used to deliver an siNA molecule or formulation of the invention locally to lung or pulmonary tissues. In another embodiment, a device comprising a nebulizer is used to deliver a an siNA molecule or formulation of the invention systemically.
H. Other Applications/Uses of siNA Molecules of the Invention
[0316] The siNA molecules of the invention can also be used for diagnostic applications, research applications, and/or manufacture of medicants.
[0317] In one aspect, the invention features a method for diagnosing a disease, trait, or condition in a subject comprising administering to the subject a composition of the invention under conditions suitable for the diagnosis of the disease, trait, or condition in the subject.
[0318] In one embodiment, siNA molecules of the invention are used to down regulate or inhibit the expression of HBV proteins arising from haplotype polymorphisms that are associated with a trait, disease or condition in a subject or organism. Analysis of HBV genes, or HBV protein or RNA levels can be used to identify subjects with such polymorphisms or those subjects who are at risk of developing traits, conditions, or diseases described herein. These subjects are amenable to treatment, for example, treatment with siNA molecules of the invention and any other composition useful in treating diseases related to target gene expression. As such, analysis of HBV protein or RNA levels can be used to determine treatment type and the course of therapy in treating a subject. Monitoring of HBV protein (e.g., hepatitis B core antigen, or HbcAg), HBV surface antigen (e.g., HBsAg) or RNA levels can be used to predict treatment outcome and to determine the efficacy of compounds and compositions that modulate the level and/or activity of certain HBV proteins associated with a trait, disorder, condition, or disease.
[0319] In another embodiment, the invention comprises use of a double-stranded nucleic acid according to the invention for use in the manufacture of a medicament. In an embodiment, the medicament is for use in treating HBV infection or a condition that is mediated by the action of HBV. In one embodiment, the medicant is for use in treating HBV infection. In some embodiments, the medicament is for use in the treatment of cancer. In a particular embodiment, the compounds of the instant invention are useful for treating hepatocellular carcinoma. In one embodiment, the medicament is for use in the treatment of liver disease. In a particular embodiment, the compounds of the instant invention are useful for treating cirrhosis of the liver.
[0320] In certain embodiments, siNAs wherein at least one strand comprises at least a 15 nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 452, SEQ ID NO: 2, SEQ ID NO: 453, SEQ ID NO: 3, SEQ ID NO: 454, SEQ ID NO: 4, or SEQ ID NO:455; or formula (A),are for use in a method for treating HBV infection.
[0321] In certain embodiments, siNAs wherein at least one strand comprises at least a 15 nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 452, SEQ ID NO: 2, SEQ ID NO: 453, SEQ ID NO: 3, SEQ ID NO: 454, SEQ ID NO: 4, or SEQ ID NO:455; or formula (A),are for use in a method for treating cancer.
[0322] In certain embodiments, siNAs wherein at least one strand comprises at least a 15 nucleotide sequence of SEQ ID NO: 1, SEQ ID NO: 452, SEQ ID NO: 2, SEQ ID NO: 453, SEQ ID NO: 3, SEQ ID NO: 454, SEQ ID NO: 4, or SEQ ID NO:455; or formula (A),are for use in a method for treating liver disease.
I. Examples
[0323] The invention will now be illustrated with the following non-limiting examples. Those of skill in the art will readily recognize a variety of non-critical parameters which can be changed or modified to yield essentially the same results.
Example 1: Design, Synthesis, and Identification of siNAs Active Against HBV.
HBVsiNA Synthesis
[0324] A series of siNA molecules were designed, synthesized and evaluated for efficacy against HBV gene expression. HBV sequences were designed and selected by selecting all possible 19 nucleotide sequences from HBV genome serotype adw2 (Accession Number X02763) and then comparing these sequences to the genome sequences of 22 other HBV subtypes shown in Table 7. 19 nucleotide sequences having perfect matches to all 22 genomes, or perfect matches in 21 of the 22 genomes and 1 or 2 mismatches to the remaining genome, were retained. These sequences were subjected to standard filters (e.g., off-target to human genome, runs of same base, matches to human miRNA seed sequences) (See also U.S. Application No. 60/182,605 for sequence design methods and standard filters). The remaining candidates were sorted by a proprietary silencing prediction score and certain sequences were selected.
[0325] The primary criteria for design for the HBV sequences for human siNAs were (i) homology between multiple HBV genotypes and serotypes and (ii) high efficacy scores as determined by a proprietary algorithm. The target sequences of the siNAs that were selected are set forth in Table la (target sequences). The sense and antisense strands of the siNA sequences corresponding to the target sequences in Table la are set forth in Table 1b. Various chemically modified siNAs that were synthesized are set forth in Table 1c.
Table la: HBV Target Sequences, noting target sites.
Target Sequence Target SiteSEQ ID NO: UCGUGGUGGACUUCUCUCA 1663 1 GUGGUGGACUUCUCUCAAU 1665 2 GCCGAUCCAUACUGCGGAA 2669 3
CCGAUCCAUACUGCGGAAC 2670 4 CAUCCUGCUGCUAUGCCUC 1818 5 UGCUGCUAUGCCUCAUCUU 1823 6 GGUGGACUUCUCUCAAUUU 1667 7 UGGUGGACUUCUCUCAAUU 1666 8 UAGACUCGUGGUGGACUUC 1658 9 UCCUCUGCCGAUCCAUACU 2663 10 UGCCGAUCCAUACUGCGGA 2668 11 UGGAUGUGUCUGCGGCGU 1783 12 CGAUCCAUACUGCGGAACU 2671 13 CGCACCUCUCUUUACGCGG 2934 14 CUGCCGAUCCAUACUGCGG 2667 15 CGUGGUGGACUUCUCUCAA 1664 16 CUGCUGCUAUGCCUCAUCU 1822 17 CCUGCUGCUAUGCCUCAUC 1821 18 CUAGACUCGUGGUGGACUU 1657 19 UCCUGCUGCUAUGCCUCAU 1820 20 GACUCGUGGUGGACUUCUC 1660 21 AUCCAUACUGCGGAACUCC 2673 22 CUCUGCCGAUCCAUACUGC 2665 23 GAUCCAUACUGCGGAACUC 2672 24 GAAGAACUCCCUCGCCUCG 567 25 AAGCCUCCAAGCUGUGCCU 54 26 AGAAGAACUCCCUCGCCUC 566 27 GGAGUGUGGAUUCGCACUC 455 28 CCUCUGCCGAUCCAUACUG 2664 29 CAAGCCUCCAAGCUGUGCC 53 30 UCCAUACUGCGGAACUCCU 2674 31 CAGAGUCUAGACUCGUGGU 1651 32 AAGAAGAACUCCCUCGCCU 565 33 GAGUGUGGAUUCGCACUCC 456 34 UCUAGACUCGUGGUGGACU 1656 35 GCUGCUAUGCCUCAUCUUC 1824 36 AGUCUAGACUCGUGGUGGA 1654 37 CUCCUCUGCCGAUCCAUAC 2662 38 UGGCUCAGUUJUACUAGUGC 2077 39 GUCUAGACUCGUGGUGGAC 1655 40 UUCAAGCCUCCAAGCUGUG 51 41 CUAUGGGAGUGGGCCUCAG 2047 42 CUCGUGGUGGACUUCUCUC 1662 43 CCUAUGGGAGUGGGCCUCA 2046 44 AGAACUCCCUCGCCUCGC 568 45 UCUGCCGAUCCAUACUGCG 2666 46
AGAGUCUAGACUCGUGGUG 1652 47 GAAGAAGAACUCCCUCGCC 564 48 UCAAGCCUCCAAGCUGUGC 52 49 AGCCUCCAAGCUGUGCCUU 55 50
[AGACUCGUGGUGGACUUCU 1659 51
Table 1b. Various HBV siNA sense and antisense sequences corresponding to the identified target sequences in Table la.
Target SEQ ID Sense Sequence Antisense Sequence SEQID Site NO: UCGUGGUGGCUUCUCUC UGAGAGAAGUCCACCACG NO: 1663 1 UCGUGGUGGACUYUCUCUCA UGAGAGAAGUCCACCACGA 452 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGUCCACCAC 453 2669 3 GCCGAUCCAUACUGCGGAA UUCCGCAGUAUGGAUCGGC 454 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUGGAUCGG 455 1818 5 CAUCCUGCUGCUAUGCCUC GAGGCAUAGCAGCAGGAUG 456 1823 6 UGCUGCUAUGCCUCAUCUU AAGAUGAGGCAUAGCAGCA 457 1667 7 GGUGGACUUCUCUCAAUUU AAAUUGAGAGAAGUCCACC 458 1666 8 UGGUGGACUUCUCUCAAUU AAUUGAGAGAAGUCCACCA 459 1658 9 UAGACUCGUGGUGGACUUC GAAGUCCACCACGAGUCUA 460 2663 10 UCCUCUGCCGAUCCAUACU AGUAUGGAUCGGCAGAGGA 461 2668 11 UGCCGAUCCAUACUGCGGA UCCGCAGUAUGGAUCGGCA 462 1783 12 UGGAUGUGUCUGCGGCGUU AACGCCGCAGACACAUCCA 463 2671 13 CGAUCCAUACUGCGGAACU AGUUCCGCAGUAUGGAUCG 464 2934 14 CGCACCUCUCUUUACGCGG CCGCGUAAAGAGAGGUGCG 465 2667 15 CUGCCGAUCCAUACUGCGG CCGCAGUAUGGAUCGGCAG 466 1664 16 CGUGGUGGACUUCUCUCAA UUGAGAGAAGUCCACCACG 467 1822 17 CUGCUGCUAUGCCUCAUCU AGAUGAGGCAUAGCAGCAG 468 1821 18 CCUGCUGCUAUGCCUCAUC GAUGAGGCAUAGCAGCAGG 469 1657 19 CUAGACUCGUGGUGGACUU AAGUCCACCACGAGUCUAG 470 1820 20 UCCUGCUGCUAUGCCUCAU AUGAGGCAUAGCAGCAGGA 471 1660 21 GACUCGUGGUGGACUUCUC GAGAAGUCCACCACGAGUC 472 2673 22 AUCCAUACUGCGGAACUCC GGAGUUCCGCAGUAUGGAU 473 2665 23 CUCUGCCGAUCCAUACUGC GCAGUAUGGAUCGGCAGAG 474 2672 24 GAUCCAUACUGCGGAACUC GAGUUCCGCAGUAUGGAUC 475 567 25 GAAGAACUCCCUCGCCUCG CGAGGCGAGGGAGUUCUUC 476 54 26 AAGCCUCCAAGCUGUGCCU AGGCACAGCUUGGAGGCUU 477 566 27 AGAAGAACUCCCUCGCCUC GAGGCGAGGGAGUUCUUCU 478 455 28 GGAGUGUGGAUUCGCACUCGAGUGCGAAUCCACACUCC 479 2664 29 CCUCUGCCGAUCCAUACUG CAGUAUGGAUCGGCAGAGG 480 53 30 CAAGCCUCCAAGCUGUGCC GGCACAGCUUGGAGGCUUG 481 2674 31 UCCAUACUGCGGAACUCCU AGGAGUUCCGCAGUAUGGA 482
1651 32 CAGAGUCUAGACUCGUGGU ACCACGAGUCUAGACUCUG 483 565 33 AAGAAGAACUCCCUCGCCU AGGCGAGGGAGUUCUUCUU 484 456 34 GAGUGUGGAUUCGCACUCC GGAGUGCGAAUCCACACUC 485 1656 35 UCUAGACUCGUGGUGGACU AGUCCACCACGAGUCUAGA 486 1824 36 GCUGCUAUGCCUCAUCUUC GAAGAUGAGGCAUAGCAGC 487 1654 37 AGUCUAGACUCGUGGUGGA UCCACCACGAGUCUAGACU 488 2662 38 CUCCUCUGCCGAUCCAUAC GUAUGGAUCGGCAGAGGAG 489 2077 39 UGGCUCAGUUUACUAGUGCGCACUAGUAAACUGAGCCA 490 1655 40 GUCUAGACUCGUGGUGGACGUCCACCACGAGUCUAGAC 491 51 41 UUCAAGCCUCCAAGCUGUG CACAGCUUGGAGGCUUGAA 492 2047 42 CUAUGGGAGUGGGCCUCAGCUGAGGCCCACUCCCAUAG 493 1662 43 CUCGUGGUGGACUUCUCUC GAGAGAAGUCCACCACGAG 494 2046 44 CCUAUGGGAGUGGGCCUCA UGAGGCCCACUCCCAUAGG 495 568 45 AAGAACUCCCUCGCCUCGC GCGAGGCGAGGGAGUUCUU 496 2666 46 UCUGCCGAUCCAUACUGCG CGCAGUAUGGAUCGGCAGA 497 1652 47 AGAGUCUAGACUCGUGGUGCACCACGAGUCUAGACUCU 498 564 48 GAAGAAGAACUCCCUCGCC GGCGAGGGAGUUCUUCUUC 499 52 49 UCAAGCCUCCAAGCUGUGC GCACAGCUUGGAGGCUUGA 500 55 50 AGCCUCCAAGCUGUGCCUU AGGCACAGCUUGGAGGCU 501 1659 51 AGACUCGUGGUGGACUUCU AGAAGUCCACCACGAGUCU 502
[0326] For each oligonucleotide of a target sequence, the two individual, complementary strands of the siNA were synthesized separately using solid phase synthesis, then purified separately by reversed phase solid phase extraction (SPE). The complementary strands were annealed to form the double strand (duplex) and delivered in the desired concentration and buffer of choice.
[0327] Briefly, the single strand oligonucleotides were synthesized using phosphoramidite chemistry on an automated solid-phase synthesizer, using procedures as are generally known in the art (see for example U.S. Application No. 12/064,014). A synthesis column was packed with solid support derivatized with the first nucleoside residue (natural or chemically modified). Synthesis was initiated by detritylation of the acid labile 5'-0-dimethoxytrityl group to release the 5'-hydroxyl. A suitably protected phosphoramidite and a suitable activator in acetonitrile were delivered simultaneously to the synthesis column resulting in coupling of the amidite to the '-hydroxyl. The column was then washed with a solvent, such as acetonitrile. An oxidizing solution, such as an iodine solution was pumped through the column to oxidize the phosphite triester linkage P(III) to its phosphotriester P(V) analog. Unreacted 5'-hydroxyl groups were capped using reagents such as acetic anhydride in the presence of 2,6-lutidine and N methylimidazole. The elongation cycle was resumed with the detritylation step for the next phosphoramidite incorporation. This process was repeated until the desired sequence was synthesized. The synthesis concluded with the final 5'-terminus protecting group (trityl or 5'-0 dimethoxytrityl).
[0328] Upon completion of the synthesis, the solid-support and associated oligonucleotide were dried under argon pressure or vacuum. Aqueous base was added and the mixture was heated to effect cleavage of the succinyl linkage, removal of the cyanoethyl phosphate protecting group, and deprotection of the exocyclic amine protection.
[0329] The following process was performed on single strands that do not contain ribonucleotides. After treating the solid support with the aqueous base, the mixture was filtered to separate the solid support from the deprotected crude synthesis material. The solid support was then rinsed with water, which is combined with the filtrate. The resultant basic solution allows for retention of the 5'-0-dimethoxytrityl group to remain on the 5' terminal position (trityl-on).
[0330] For single strands that contain ribonucleotides, the following process was performed. After treating the solid support with the aqueous base, the mixture was filtered to separate the solid support from the deprotected crude synthesis material. The solid support was then rinsed with dimethylsulfoxide (DMSO), which was combined with the filtrate. Fluoride reagent, such as triethylamine trihydrofluoride, was added to the mixture, and the solution was heated. The reaction was quenched with suitable buffer to provide a solution of crude single strand with the '-O-dimethoxytrityl group on the final 5' terminal position.
[0331] The trityl-on solution of each crude single strand was purified using chromatographic purification, such as SPE RPC purification. The hydrophobic nature of the trityl group permits stronger retention of the desired full-length oligo than the non-tritylated truncated failure sequences. The failure sequences were selectively washed from the resin with a suitable solvent, such as low percent acetonitrile. Retained oligonucleotides were then detritylated on-column with trifluoroacetic acid to remove the acid-labile trityl group. Residual acid was washed from the column, a salt exchange was performed, and a final desalting of the material commenced .
The full-length oligo was recovered in a purified form with an aqueous-organic solvent. The final product was then analyzed for purity (HPLC), identity (Maldi-TOF MS), and yield (UV A2 6 0). The oligos were dried via lyophilization or vacuum condensation.
[0332] Annealing: Based on the analysis of the product, the dried oligos were dissolved in appropriate buffers followed by mixing equal molar amounts (calculated using the theoretical extinction coefficient) of the sense and antisense oligonucleotide strands. The solution was then analyzed for purity of duplex by chromatographic methods and desired final concentration. If the analysis indicated an excess of either strand, then the additional non-excess strand was titrated until duplexing was complete. When analysis indicated that the target product purity has been achieved the material was delivered and ready for use.
[0333] Below is a table showing various modified siNAs synthesized using this protocol or that can be synthesized using this protocol or using methods known in the art.
Table 1c: HBV siNA Strands Synthesized (Antisense sequences are readily identified as having complete or partial complementarity to the target seque ce shown). SEQ SEQ siNA duplex ID Site ID Target Sequence Modified Sequence ID NO: NO: R-008351268-000C 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCATT B 52 R-008351268-000C 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGUCCACCACGAUU 53 R-008351278-000V 1665 2 GUGGUGGACUUCUCUCAAUB GuGGuGGAcuucucucAAuTTB 54 R-008351278-000V 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGuccAccAcUU 55 R-008351342-000A 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAATT B 56 R-008351342-000A 2669 3 GCCGAUCCAUACUGCGGAA UUCCGCAGUAUGGAUCGGCUU 57 R-008351172-000H 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUTT B 58 R-008351172-000H 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGUCCACCACUU 59 R-008351362-000K 2670 4 CCGAUCCAUACUGCGGAAC B ccGAuccAuAcuGcGGAAcTTB 60 R-008351362-000K 2670 4 CCGAUCCAUACUGCGGAAC GUUccGcAGuAuGGAucGGUU 61 R-008351389-000F 1818 5 CAUCCUGCUGCUAUGCCUC B CAUCCUGCUGCUAUGCCUCTTB 62 R-008351389-000F 1818 5 CAUCCUGCUGCUAUGCCUC GAGGCAUAGCAGCAGGAUGUU 63 R-008351306-000R 1823 6 UGCUGCUAUGCCUCAUCUU B UGCUGCUAUGCCUCAUCUUTTB 64 R-008351306-000R 1823 6 UGCUGCUAUGCCUCAUCUU AAGAUGAGGCAUAGCAGCAUU 65 R-008351372-000C 1667 7 GGUGGACUUCUCUCAAUUU B GGuGGAcuucucucAAuuuTT B 66 R-008351372-000C 1667 7 GGUGGACUUCUCUCAAUUU AAAuuGAGAGAAGuccAccUU 67 R-008351317-000S 1666 8 UGGUGGACUUCUCUCAAUU B uGGuGGAcuucucucAAuuTT B 68 R-008351317-000S 1666 8 UGGUGGACUUCUCUCAAUU AAUuGAGAGAAGuccAccAUU 69 R-008351210-000W 1658 9 UAGACUCGUGGUGGACUUC B UAGACUCGUGGUGGACUUCTT B 70 R-008351210-000W 1658 9 UAGACUCGUGGUGGACUUC GAAGUCCACCACGAGUCUAUU 71 R-008351329-000B 2663 10 UCCUCUGCCGAUCCAUACU BUCCUCUGCCGAUCCAUACUTTB 72 R-008351329-000B 2663 10 UCCUCUGCCGAUCCAUACU AGUAUGGAUCGGCAGAGGAUU 73 R-008351350-000A 1658 9 UAGACUCGUGGUGGACUUCB uAGAcucGuGGuGGAcuucTTB 74
R-008351350-OOOA 1658 9 UAGACUCGUGGUGGACUUC GAAGuccAccAcGAGucuAUU 75 R-008351303-OOOP 2668 11 UGCCGAUCCAUACUGCGGA B UGCCGAUCCAUACUGCGGATT B 76 R-008351303-OOOP 2668 11 UGCCGAUCCAUACUGCGGA UCCGCAGUAUGGAUCGGCAUU 77 R-008351229-OOOS 1783 12 UGGAUGUGUCUGCGGCGUU B UGGAUGUGUCUGCGGCGUUTT B 78 R-008351229-OOOS 1783 12 UGGAUGUGUCUGCGGCGUU AACGCCGCAGACACAUCCAUU 79 R-008351386-OOOE 2671 13 CGAUCCAUACUGCGGAACU B CGAUCCAUACUGCGGAACUTTB 80 R-008351386-OOOE 2671 13 CGAUCCAUACUGCGGAACU AGUUCCGCAGUAUGGAUCGUU 81 R-008351300-OOON 2934 14 CGCACCUCUCUUUACGCGG B CGCACCUCUCUUUACGCGGTTB 82 R-008351300-OOON 2934 14 CGCACCUCUCUUUACGCGG CCGCGUAAAGAGAGGUGCGUU 83 R-008351263-000J 2667 15 CUGCCGAUCCAUACUGCGG B CUGCCGAUCCAUACUGCGGTTB 84 R-008351263-000J 2667 15 CUGCCGAUCCAUACUGCGG CCGCAGUAUGGAUCGGCAGUU 85 R-008351190-OOOA 1783 12 UGGAUGUGUCUGCGGCGU B uGGAuGuGucuGcGGcGuuTTB 86 R-008351190-OOOA 1783 12 UGGAUGUGUCUGCGGCGUU AACGccGcAGAcAcAuccAUU 87 R-008351326-OOOA 1666 8 UGGUGGACUUCUCUCAAUU B UGGUGGACUUCUCUCAAUUTT B 88 R-008351326-OOOA 1666 8 UGGUGGACUUCUCUCAAUU AAUUGAGAGAAGUCCACCAUU 89 R-008351260-OOOH 1664 16 CGUGGUGGACUUCUCUCAA B CGUGGUGGACUUCUCUCAATT B 90 R-008351260-OOOH 1664 16 CGUGGUGGACUUCUCUCAA UGAGAGAAGUCCACCACGUU 91 R-008351195-OOOU 1822 17 CUGCUGCUAUGCCUCAUCU B CUGCUGCUAUGCCUCAUCUTTB 92 R-008351195-OOOU 1822 17 CUGCUGCUAUGCCUCAUCU AGAUGAGGCAUAGCAGCAGUU 93 R-008351323-OOOZ 1821 18 CCUGCUGCUAUGCCUCAUC B CCUGCUGCUAUGCCUCAUCTTB 94 R-008351323-OOOZ 1821 18 CCUGCUGCUAUGCCUCAUC GAUGAGGCAUAGCAGCAGGUU 95 R-008351367-OOOD 1667 7 GGUGGACUUCUCUCAAUUU B GGUGGACUUCUCUCAAUUUTT B 96 R-008351367-000D 1667 7 GGUGGACUUCUCUCAAUUU AAAUUGAGAGAAGUCCACCUU 97 R-008351251-OOOZ 1657 19 CUAGACUCGUGGUGGACUU B cuA GAcucGuGGuGGAcuuTT B 98 R-008351251-OOOZ 1657 19 CUAGACUCGUGGUGGACUU AAGuccAccAcGAGucuAGUU 99 R-008351240-OOOY 1820 20 UCCUGCUGCUAUGCCUCAU B UCCUGCUGCUAUGCCUCAUTTB 100 R-008351240-OOOY 1820 20 UCCUGCUGCUAUGCCUCAU AUGAGGCAUAGCAGCAGGAUU 101 R-008351178-000K 2668 11 UGCCGAUCCAUACUGCGGA B uGccGAuccAuAcuGcGGATTB 102 R-008351178-000K 2668 11 UGCCGAUCCAUACUGCGGA UCCGcAGuAuGGAucGGcAUU 103 R-008351353-OOOB 1657 19 CUAGACUCGUGGUGGACUUB CUAGACUCGUGGUGGACUUTTB 104 R-008351353-OOOB 1657 19 CUAGACUCGUGGUGGACUU AAGUCCACCACGAGUCUAGUU 105 R-008351297-OOOW 1660 21 GACUCGUGGUGGACUUCUC B GACUCGUGGUGGACUUCUCTTB 106 R-008351297-OOOW 1660 21 GACUCGUGGUGGACUUCUC GAGAAGUCCACCACGAGUCUU 107 R-008351175-000J 2673 22 AUCCAUACUGCGGAACUCC B AuccAuAcuGGGAcuccTT B 108 R-008351175-000J 2673 22 AUCCAUACUGCGGAACUCC GGAGuuccGcAGuAuGGAuUU 109 R-008351181-OOOS 2665 23 CUCUGCCGAUCCAUACUGC B cucuGccGAuccAuAcuGcTTB 110 R-008351181-OOOS 2665 23 CUCUGCCGAUCCAUACUGC GCAGuAuGGAucGGcAGAGUU 111 R-008351184-000T 2672 24 GAUCCAUACUGCGGAACUC B GAUCCAUACUGCGGAACUCTTB 112 R-008351184-OOOT 2672 24 GAUCCAUACUGCGGAACUC GAGUUCCGCAGUAUGGAUCUU 113 R-008351187-OOOU 567 25 GAAGAACUCCCUCGCCUCG B GAAGAACUCCCUCGCCUCGTTB 114 R-008351187-OOOU 567 25 GAAGAACUCCCUCGCCUCG CGAGGCGAGGGAGUUCUUCUU 115 R-008351192-OOOT 54 26 AGCCUCCAAGCUGUGCCU B AAGccuccAAGcuGuGccuTT B 116 R-008351192-000T 54 26 AAGCCUCCAAGCUGUGCCU AGGcAcAGcuuGGAGGcuuUU 117 R-008351198-OOOV 566 27 AGAAGAACUCCCUCGCCUC B AGAAGAACUCCCUCGCCUCTT B 118 R-008351198-OOOV 566 27 AGAAGAACUCCCUCGCCUC GAGGCGAGGGAGUUCUUCUUU 119 R-008351201-OOOM 2671 13 CGAUCCAUACUGCGGAACU B cGAuccAuAcuGcGGAAcuTTB 120 R-008351201-OOOM 2671 13 CGAUCCAUACUGCGGAACU AGUuccGcAGuAuGGAucGUU 121 R-008351204-OOON 455 28 GGAGUGUGGAUUCGCACUC B GGAGUGUGGAUUCGCACUCTT B 122 R-008351204-OOON 455 28 GGAGUGUGGAUUCGCACUC GAGUGCGAAUCCACACUCCUU 123 R-008351207-OOOP 2664 29 CCUCUGCCGAUCCAUACUG B cucuGcGAuccAuAcuGTT B 124 R-008351207-OOOP 2664 29 CCUCUGCCGAUCCAUACUG CAGuAuGGAucGGcAGAGGUU 125
R-008351212-OOON 53 30 CAAGCCUCCAAGCUGUGCC B cAAGccuccAAGcuGuGccTT B 126 R-008351212-OOON 53 30 CAAGCCUCCAAGCUGUGCC GGCAcAGcuuGGAGGcuuGUU 127 R-008351215-OOOP 2674 31 UCCAUACUGCGGAACUCCU B UCCAUACUGCGGAACUCCUTTB 128 R-008351215-OOOP 2674 31 UCCAUACUGCGGAACUCCU AGGAGUUCCGCAGUAUGGAUU 129 R-008351218-OOOR 1651 32 CAGAGUCUAGACUCGUGGU B CAGAGUCUAGACUCGUGGUTT B 130 R-008351218-OOOR 1651 32 CAGAGUCUAGACUCGUGGU ACCACGAGUCUAGACUCUGUU 131 R-008351220-OOON 565 33 AAGAAGAACUCCCUCGCCU BAAGAAGAAcucccucGccuTTB 132 R-008351220-OOON 565 33 AAGAAGAACUCCCUCGCCU AGGcGAGGGAGuucuucuuUU 133 R-008351223-OOOP 456 34 GAGUGUGGAUUCGCACUCC B GAGUGUGGAUUCGCACUCCTTB 134 R-008351223-OOOP 456 34 GAGUGUGGAUUCGCACUCC GGAGUGCGAAUCCACACUCUU 135 R-008351226-OOOR 1656 35 UCUAGACUCGUGGUGGACU B UCUAGACUCGUGGUGGACUTT B 136 R-008351226-OOOR 1656 35 UCUAGACUCGUGGUGGACU AGUCCACCACGAGUCUAGAUU 137 R-008351232-OOOY 2663 10 UCCUCUGCCGAUCCAUACU B uccucuGcGAuccAuAcuTT B 138 R-008351232-OOOY 2663 10 UCCUCUGCCGAUCCAUACU AGUAuGGAucGGcAGAGGAUU 139 R-008351235-OOOZ 53 30 CAAGCCUCCAAGCUGUGCC B CAAGCCUCCAAGCUGUGCCTTB 140 R-008351235-OOOZ 53 30 CAAGCCUCCAAGCUGUGCC GGCACAGCUUGGAGGCUUGUU 141 R-008351237-OOOS 1821 18 CCUGCUGCUAUGCCUCAUC B ccuGcuGcuuGccucAucTT B 142 R-008351237-000S 1821 18 CCUGCUGCUAUGCCUCAUC GAUGAGGcAuAGcAGcAGGUU 143 R-008351243-000Z 2665 23 CUCUGCCGAUCCAUACUGCB CUCUGCCGAUCCAUACUGCTTB 144 R-008351243-000Z 2665 23 CUCUGCCGAUCCAUACUGC GCAGUAUGGAUCGGCAGAGUU 145 R-008351246-000A 1651 32 CAGAGUCUAGACUCGUGGUB cAGAGucuAGAcucGuGGuTTB 146 R-008351246-000A 1651 32 CAGAGUCUAGACUCGUGGU ACCAcGAGucuAGAcucuGUU 147 R-008351248-000T 1824 36 GCUGCUAUGCCUCAUCUUC B GcuGcuAuGccucAucuucTTB 148 R-008351248-000T 1824 36 CUGCUAUGCCUCAUCUUC GAAGAuGAGGcAuAGcAGcUU 149 R-008351254-000A 2664 29 CCUCUGCCGAUCCAUACUGB CCUCUGCCGAUCCAUACUGTTB 150 R-008351254-OOOA 2664 29 CCUCUGCCGAUCCAUACUG CAGUAUGGAUCGGCAGAGGUU 151 R-008351257-OOOB 1656 35 UCUAGACUCGUGGUGGACUB ucuAGAcucGuGGuGGAcuTTB 152 R-008351257-OOOB 1656 35 UCUAGACUCGUGGUGGACU AGUccAccAcGAGucuAGAUU 153 R-008351266-OOOK 1818 5 CAUCCUGCUGCUAUGCCUC B cAuccuGcuGcuAuGccucTT B 154 R-008351266-OOOK 1818 5 CAUCCUGCUGCUAUGCCUC GAGGcAuAGcAGcAGGAuGUU 155 R-008351271-000J 1654 37 AGUCUAGACUCGUGGUGGA BAGucuA GAcucGuGGuGGATTB 156 R-008351271-000J 1654 37 AGUCUAGACUCGUGGUGGA UCCAccAcGAGucuAGAcuUU 157 R-008351274-000K 455 28 GGAGUGUGGAUUCGCACUC B GGA GuGuGGAuucGcAcucTT B 158 R-008351274-000K 455 28 GGAGUGUGGAUUCGCACUC GAGuGcGAAuccAcAcuccUU 159 R-008351276-OOOC 1823 6 UGCUGCUAUGCCUCAUCUU B uGcuGcuAuGccucAucuuTTB 160 R-008351276-OOOC 1823 6 UGCUGCUAUGCCUCAUCUU AAGAuGAGGcAuAGcAGcAUU 161 R-008351281-OOOB 2674 31 UCCAUACUGCGGAACUCCU BuccAuAcuGcGGAAcuccuTTB 162 R-008351281-OOOB 2674 31 UCCAUACUGCGGAACUCCU AGGAGuuccGcAGuAuGGAUU 163 R-008351284-OOOC 2662 38 CUCCUCUGCCGAUCCAUAC BCUCCUCUGCCGAUCCAUACTTB 164 R-008351284-OOOC 2662 38 CUCCUCUGCCGAUCCAUAC GUAUGGAUCGGCAGAGGAGUU 165 R-008351287-OOOD 2672 24 GAUCCAUACUGCGGAACUC B GAuccAuAcuGcGGAAcucTTB 166 R-008351287-OOOD 2672 24 GAUCCAUACUGCGGAACUC GAGuuccGcAGuAuGGAucUU 167 R-008351290-OOOK 567 25 GAAGAACUCCCUCGCCUCG B GAAGAAcucccucGccucGTTB 168 R-008351290-000K 567 25 GAAGAACUCCCUCGCCUCG CGAGGcGAGGGAGuucuucUU 169 R-008351292-OOOC 1663 1 UCGUGGUGGACUUCUCUCA B ucGuGGuGGAcuucucucATTB 170 R-008351292-OOOC 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGuccAccAcGAUU 171 R-008351294-OOOV 1820 20 UCCUGCUGCUAUGCCUCAU B uccuGcuGcuAuGccucAuTTB 172 R-008351294-OOOV 1820 20 UCCUGCUGCUAUGCCUCAU AUGAGGcAuAGcAGcAGGAUU 173 R-008351309-OOOS 2673 22 AUCCAUACUGCGGAACUCC B AUCCAUACUGCGGAACUCCTTB 174 R-008351309-OOOS 2673 22 AUCCAUACUGCGGAACUCC GGAGUUCCGCAGUAUGGAUUU 175 R-008351312-OOOY 2077 39 UGGCUCAGUUUACUAGUGC B UGGCUCAGUUUACUAGUGCTT B 176
R-008351312-OOOY 2077 39 UGGCUCAGUUUACUAGUGC GCACUAGUAAACUGAGCCAUU 177 R-008351315-OOOZ 54 26 AAGCCUCCAAGCUGUGCCU B AAGCCUCCAAGCUGUGCCUTTB 178 R-008351315-OOOZ 54 26 AAGCCUCCAAGCUGUGCCU AGGCACAGCUUGGAGGCUUUU 179 R-008351320-OOOY 566 27 AGAAGAACUCCCUCGCCUC BAGAAGAAcucccucGccucTTB 180 R-008351320-OOOY 566 27 AGAAGAACUCCCUCGCCUC GAGGcGAGGGAGuucuucuUU 181 R-008351332-OOOH 1655 40 GUCUAGACUCGUGGUGGAC B GucuA GAcucGuGGuGGAcTT B 182 R-008351332-OOOH 1655 40 GUCUAGACUCGUGGUGGAC GUCcAccAcGAGucuAGAcUU 183 R-008351334-OOOA 1822 17 CUGCUGCUAUGCCUCAUCU B cuGcuGcuAuGccucAucuTTB 184 R-008351334-OOOA 1822 17 CUGCUGCUAUGCCUCAUCU AGAuGAGGcAuAGcAGcAGUU 185 R-008351337-OOOB 2667 15 CUGCCGAUCCAUACUGCGG B cuGcGAuccAuAcuGcGGTTB 186 R-008351337-OOOB 2667 15 CUGCCGAUCCAUACUGCGG CCGcAGuAuGGAucGGcAGUU 187 R-008351339-OOOU 51 41 UUCAAGCCUCCAAGCUGUG BuucAAGccuccAAGcuGuGTTB 188 R-008351339-OOOU 51 41 UUCAAGCCUCCAAGCUGUG CACAGcuuGGAGGcuuGAAUU 189 R-008351345-OOOB 1654 37 AGUCUAGACUCGUGGUGGA B AGUCUAGACUCGUGGUGGATT B 190 R-008351345-OOOB 1654 37 AGUCUAGACUCGUGGUGGA UCCACCACGAGUCUAGACUUU 191 R-008351348-OOOC 1655 40 GUCUAGACUCGUGGUGGAC B GUCUAGACUCGUGGUGGACTT B 192 R-008351348-000C 1655 40 GUCUAGACUCGUGGUGGAC GUCCACCACGAGUCUAGACUU 193 R-008351356-OOOC 2047 42 CUAUGGGAGUGGGCCUCAGB CUAUGGGAGUGGGCCUCAGTTB 194 R-008351356-OOOC 2047 42 CUAUGGGAGUGGGCCUCAGCUGAGGCCCACUCCCAUAGUU 195 R-008351359-OOOD 51 41 UUCAAGCCUCCAAGCUGUG B UUCAAGCCUCCAAGCUGUGTTB 196 R-008351359-OOOD 51 41 UUCAAGCCUCCAAGCUGUG CACAGCUUGGAGGCUUGAAUU 197 R-008351364-000C 1660 21 GACUCGUGGUGGACUUCUC B GAcucGuGGuGGAcuucucTT B 198 R-008351364-000C 1660 21 GACUCGUGGUGGACUUCUC GAGAAGuccAccAcGAGucUU 199 R-008351370-OOOK 1824 36 GCUGCUAUGCCUCAUCUUC B GCUGCUAUGCCUCAUCUUCTTB 200 R-008351370-OOOK 1824 36 GCUGCUAUGCCUCAUCUUC GAAGAUGAGGCAUAGCAGCUU 201 R-008351374-OOOV 1664 16 CGUGGUGGACUUCUCUCAA B cGuGGuGGAcuucucucAATT B 202 R-008351374-OOOV 1664 16 CGUGGUGGACUUCUCUCAA UGAGAGAAGuccAccAcGUU 203 R-008351377-OOOW 2077 39 UGGCUCAGUUUACUAGUGC B uGGcucAGuuuAcuAGuGcTT B 204 R-008351377-000W 2077 39 UGGCUCAGUUUACUAGUGC GCAcuAGuAAAcuGAGccAUU 205 R-008351380-OOOC 2662 38 CUCCUCUGCCGAUCCAUAC B cuccucuGcGAuccAuAcTT B 206 R-008351380-OOOC 2662 38 CUCCUCUGCCGAUCCAUAC GUAuGGAucGGcAGAGGAGUU 207 R-008351383-OOOD 456 34 GAGUGUGGAUUCGCACUCC B GAGuGuGGAuucGcAcuccTT B 208 R-008351383-OOOD 456 34 GAGUGUGGAUUCGCACUCC GGAGuGcGAAuccAcAcucUU 209 R-008351392-OOOM 2934 14 CGCACCUCUCUUUACGCGG B cGcAccucucuuuAcGcGGTTB 210 R-008351392-OOOM 2934 14 CGCACCUCUCUUUACGCGG CCGcGuAAAGAGAGGuGcGUU 211 R-008351395-OOON 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACTTB 212 R-008351395-OOON 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUGGAUCGGUU 213 R-008351398-OOOP 2047 42 CUAUGGGAGUGGGCCUCAGBcuAuGGGAGuGGGccucAGTTB 214 R-008351398-OOOP 2047 42 CUAUGGGAGUGGGCCUCAG CUGAGGcccAcucccAuAGUU 215 R-008351401-OOOG 565 33 AAGAAGAACUCCCUCGCCU B AAGAAGAACUCCCUCGCCUTTB 216 R-008351401-OOOG 565 33 AAGAAGAACUCCCUCGCCU AGGCGAGGGAGUUCUUCUUUU 217 R-008351404-OOOH 2669 3 GCCGAUCCAUACUGCGGAA B GccGAuccAuAcuGcGGAA TTB 218 R-008351404-OOOH 2669 3 GCCGAUCCAUACUGCGGAA UCcGcAGuAuGGAucGGcUU 219 R-008380305-OOOE 2670 4 CCGAUCCAUACUGCGGAAC B ccGAuccAuAcuGcGGAAcUsU B 220 R-008380305-OOOE 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUGGAUCGGUsU 221 R-008380308-OOOF 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 222 R-008380308-OOOF 2670 4 CCGAUCCAUACUGCGGAAC GuuccGcAGuAuGGAucGGUsU 223 R-008380311-OOOM 2669 3 CCGAUCCAUACUGCGGAA B GccGAuccAuAcuGcGGAAUsU B 224 R-008380311-OOOM 2669 3 CCGAUCCAUACUGCGGAA UUCCg!gaUggaUlggCUsg 225 R-008380313-OOOE 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 226 R-008380313-OOOE 2669 3 CCGAUCCAUACUGCGGAA UCCg!agUagggaUlggCUsU 225
R-008380315-OOOX 2669 3 GCCGAUCCAUACUGCGGAA B GccGAuccAuAcuGcGGAAUsU B 224 R-008380315-OOOX 2669 3 GCCGAUCCAUACUGCGGAA UCCgCagUaUggaUCggCUsU 227 R-008380316-000F 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 226 R-008380316-OOOF 2669 3 GCCGAUCCAUACUGCGGAA UCCgCagUaUggaUCggCUsU 227 R-008380318-OOOY 2669 3 CCGAUCCAUACUGCGGAA uuccGcAGuAuGGAucGGcUsU 228 R-008380318-OOOY 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 226 R-008380321-OOOE 2669 3 GCCGAUCCAUACUGCGGAA B GccGAuccAuAcuGcGGAAUsU B 229 R-008380321-OOOE 2669 3 GCCGAUCCAUACUGCGGAA UUCcGcAGuAuGGAucGGcUsU 230 R-008380324-OOOF 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 231 R-008380324-000F 2669 3 GCCGAUCCAUACUGCGGAA UUCCGCAGUAUGGAUCGGCUsU 232 R-008380327-000G 1665 2 GUGGUGGACUUCUCUCAAU B GuGGuGGAcuucucucAAuUsU B 233 R-008380327-000G 1665 2 GUGGUGGACUUCUCUCAAU AuuGAGAGAAGuccAccAcUsU 234 R-008380330-000N 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 235 R-008380330-000N 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGUCCACCACUsU 236 R-008380333-000P 1663 1 UCGUGGUGGACUUCUCUCA B ucGuGGuGGAcuucucucAUsU B 237 R-008380333-000P 1663 1 UCGUGGUGGACUUCUCUCA UGAgagaagUCCaC CgaUsU 238 R-008380335-000G 1663 1 UCGUGGUGGACUUCUCUCA B ucGuGGuGGAcuucucucAUsU B 237 R-008380335-000G 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGuccAccAcGAUsU 239 R-008380337-000Z 1663 1 UCGUGGUGGACUUCUCUCA B ucGuGGuGGAcuucucucAUsU B 237 R-008380337-000Z 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGUCCACCACGAUsU 240 R-008380340-000F 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 241 R-008380340-000F 1663 1 UCGUGGUGGACUUCUCUCA UGAgagaagUCCaCCaCgaUsU 242 R-008380343-000G 1663 1 UCGUGGUGGACUUCUCUCA uGAGAGAAGuccAccAcGAUsU 243 R-008380343-000G 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 244 R-008380346-000H 2670 4 CCGAUCCAUACUGCGGAAC B ccGAuccAuAcuGcGGAAcUsU B 245 R-008380346-000H 2670 4 CCGAUCCAUACUGCGGAAC GUUccGcAGuAuGGAucGGUsU 246 R-008380348-000A 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 247 R-008380348-000A 2669 3 GCCGAUCCAUACUGCGGAA UUCCg!agUjagggaUlggCUsg 225 R-008380349-000J 2669 3 GCCGAUCCAUACUGCGGAA B GccGAuccAuAcuGcGGAAUsU B 224 R-008380349-000J 2669 3 GCCGAUCCAUACUGCGGAA UUCcGcAGuAuGGAucGGcUsU 230 R-008380351-000G 2669 3 GCCGAUCCAUACUGCGGAA B GccGAuccAuAcuGcGGAAUsU B 224 R-008380351-OOOG 2669 3 GCCGAUCCAUACUGCGGAA UUCCGCAGUAUGGAUCGGCUsU 248 R-008380352-OOOR 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 247 R-008380352-OOOR 2669 3 GCCGAUCCAUACUGCGGAA UUCCGCAGUAUGGAUCGGCUsU 248 R-008380353-OOOZ 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 247 R-008380353-OOOZ 2669 3 GCCGAUCCAUACUGCGGAA UCCgCagUaUggaUCggCUsU 227 R-008380354-OOOH 2669 3 GCCGAUCCAUACUGCGGAA uuccGcAGuAuGGAucGGcUsU 228 R-008380354-OOOH 2669 3 GCCGAUCCAUACUGCGGAA B GccGAuccAuAcuGcGGAAUsU B 224 R-008380356-OOOA 2669 3 GCCGAUCCAUACUGCGGAA uuccGcAGuAuGGAucGGcUsU 228 R-008380356-OOOA 2669 3 GCCGAUCCAUACUGCGGAA B GccGAuccAuAcuGcGGAAUsU B 249 R-008380359-OOOB 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 250 R-008380359-OOOB 1665 2 GUGGUGGACUUCUCUCAAU AUUgagagaagUCCaCCaCUsU 251 R-008380361-OOOZ 1665 2 GUGGUGGACUUCUCUCAAU B GuGGuGGAcuucucucAAuUsU B 252 R-008380361-OOOZ 1665 2 GUGGUGGACUUCUCUCAAU AuuGAGAGAAGuccAccAcUsU 234 R-008380362-OOOH 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 241 R-008380362-OOOH 1663 1 UCGUGGUGGACUUCUCUCA UGAgagaagUCCaC CgaUsU 238 R-008380363-OOOS 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 241 R-008380363-OOOS 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGUCCACCACGAUsU 240 R-008380365-000J 1663 1 UCGUGGUGGACUUCUCUCA uGAGAGAAGuccAccAcGAUsU 243 R-008380365-000J 1663 1 UCGUGGUGGACUUCUCUCA B ucGuGGuGGAcuucucucAUsU B 253 R-008380367-OOOB 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 222
R-008380367-OOOB 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUGGAUCGGUsU 254 R-008380369-OOOU 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 222 R-008380369-OOOU 2670 4 CCGAUCCAUACUGCGGAAC GUUCCgCagUaUggaUCggUisU 255 R-008380370-OOOH 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 226 R-008380370-OOOH 2669 3 CCGAUCCAUACUGCGGAA UUCCGCAGUAUGGAUCGGCUsU 248 R-008380372-OOOA 1665 2 GUGGUGGACUUCUCUCAAU B GuGGuGGAcuucucucAAuUsU B 233 R-008380372-OOOA 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGUCCACCACUsU 256 R-008380374-OOOT 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 257 R-008380374-000T 1665 2 GUGGUGGACUUCUCUCAAU AUUgagagaagUCCaCCaCUsU 251 R-008380377-OOOU 1665 2 GUGGUGGACUUCUCUCAAU B GuGGuGGAcuucucucAAuUsU B 258 R-008380377-OOOU 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGuccAccAcUsU 259 R-008380378-OOOC 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 244 R-008380378-000C 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGuccAccAcGAUsU 239 R-008380379-000L 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 241 R-008380379-000L 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGuccAccAcGAUsU 239 R-008380381-000J 1663 1 UCGUGGUGGACUUCUCUCA B ucGuGGuGGAcuucucucAUsU B 237 R-008380381-000J 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGUCCACCACGAUsU 260 R-008380383-OOOB 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 261 R-008380383-OOOB 2670 4 CCGAUCCAUACUGCGGAAC GUUCCgCagUaUggaUCggUsU 255 R-008380384-OOOK 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 261 R-008380384-000K 2670 4 CCGAUCCAUACUGCGGAAC GuuccGcAGuAuGGAucGGUsU 223 R-008380386-OOOC 2670 4 CCGAUCCAUACUGCGGAAC B ccGAuccAuAcuGcGGAAcUsU B 262 R-008380386-OOOC 2670 4 CCGAUCCAUACUGCGGAAC GuuccGcAGuAuGGAucGGUsU 223 R-008380387-OOOL 2669 3 GCCGAUCCAUACUGCGGAA uuccGcAGuAuGGAucGGcUsU 228 R-008380387-OOOL 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 247 R-008380389-OOOD 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 250 R-008380389-OOOD 1665 2 GUGGUGGACUUCUCUCAAU AUUgagagaagUCCaCCaCUsU 263 R-008380390-000T 1665 2 GUGGUGGACUUCUCUCAAU B GuGGuGGAcuucucucAAuUsU B 233 R-008380390-000T 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGuccAccAcUsU 259 R-008380391-OOOB 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 250 R-008380391-OOOB 1665 2 GUGGUGGACUUCUCUCAAU AuuGAGAGAAGuccAccAcUsU 234 R-008380394-OOOC 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 264 R-008380394-OOOC 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGUCCACCACGAUsU 265 R-008380396-OOOV 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 222 R-008380396-OOOV 2670 4 CCGAUCCAUACUGCGGAAC GtUGg ~agUaUggaUggUsU_ 266 R-008380397-OOOD 2670 4 CCGAUCCAUACUGCGGAAC B ccGAuccAuAcuGcGGAAcUsU B 220 R-008380397-OOOD 2670 4 CCGAUCCAUACUGCGGAAC GUUccGcAGuAuGGAucGGUsU 246 R-008380398-OOOM 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 261 R-008380398-OOOM 2670 4 CCGAUCCAUACUGCGGAAC GUUccGcAGuAuGGAucGGUsU 246 R-008380399-OOOW 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 222 R-008380399-OOOW 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUGGAUCGGUsU 221 R-008380401-OOOE 2669 3 GCCGAUCCAUACUGCGGAA B GccGAuccAuAcuGcGGAAUsU B 224 R-008380401-OOOE 2669 3 GCCGAUCCAUACUGCGGAA UUCCGCAGUAUGGAUCGGCUsU 267 R-008380402-OOON 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 250 R-008380402-OOON 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGuccAccAcUsU 259 R-008380404-OOOF 1665 2 GUGGUGGACUUCUCUCAAU B GuGGuGGAcuucucucAAuUsU B 233 R-008380404-OOOF 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGUCCACCACUsU 268 R-008380405-OOOP 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 257 R-008380405-OOOP 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGUCCACCACUsU 268 R-008380406-OOOY 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 250 R-008380406-OOOY 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGUCCACCACUsU 268
R-008380407-OOOG 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 241 R-008380407-OOOG 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGUCCACCACGAUsU 260 R-008380408-OOOR 1663 1 UCGUGGUGGACUUCUCUCA uGAGAGAAGuccAccAcGAUsU 243 R-008380408-OOOR 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 241 R-008380409-OOOZ 2670 4 CCGAUCCAUACUGCGGAAC B ccGAuccAuAcuGcGGAAcUsU B 220 R-008380409-OOOZ 2670 4 CCGAUCCAUACUGCGGAAC GUUGCg ~agUaUggaUggUsU_ 266 R-008380410-000N 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 222 R-008380410-000N 2670 4 CCGAUCCAUACUGCGGAAC GUUccGcAGuAuGGAucGGUsU 246 R-008380411-OOOX 2670 4 CCGAUCCAUACUGCGGAAC B ccGAuccAuAcuGcGGAAcUsU B 220 R-008380411-000X 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUGGAUCGGUsU 254 R-008380412-000F 2670 4 CCGAUCCAUACUGCGGAAC B ccGAuccAuAcuGcGGAAcUsU B 220 R-008380412-OOOF 2670 4 CCGAUCCAUACUGCGGAAC GUUCCgCagUaUggaUCggUsU 255 R-008380413-OOOP 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 247 R-008380413-OOOP 2669 3 GCCGAUCCAUACUGCGGAA UUCcGcAGuAuGGAucGGcUsU 230 R-008380414-OOOY 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 226 R-008380414-OOOY 2669 3 GCCGAUCCAUACUGCGGAA UUCCGCAGUAUGGAUCGGCUsU 267 R-008380415-OOOG 1665 2 GUGGUGGACUUCUCUCAAU B GuGGuGGAcuucucucAAuUsU B 233 R-008380415-OOOG 1665 2 GUGGUGGACUUCUCUCAAU AUUgagagaagUCCaCCaCUsU 263 R-008380416-OOOR 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 257 R-008380416-OOOR 1665 2 GUGGUGGACUUCUCUCAAU AUUgagagaagUCCaCCaCUsU 263 R-008380417-OOOZ 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 250 R-008380417-OOOZ 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGUCCACCACUsU 256 R-008380418-OOOH 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 244 R-008380418-OOOH 1663 1 UCGUGGUGGACUUCUCUCA UGAgagaagUCCaCCaCgaUsU 242 R-008380420-OOOF 1663 1 UCGUGGUGGACUUCUCUCA B ucGuGGuGGAcuucucucAUsU B 269 R-008380420-OOOF 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGuccAccAcGAUsU 239 R-008380421-OOOP 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 261 R-008380421-OOOP 2670 4 CCGAUCCAUACUGCGGAAC GtUGg ~agUaUggaUCggUsU_ 266 R-008380422-OOOY 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 261 R-008380422-OOOY 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUGGAUCGGUsU 254 R-008380423-OOOG 2670 4 CCGAUCCAUACUGCGGAAC B ccGAuccAuAcuGcGGAAcUsU B 220 R-008380423-OOOG 2670 4 CCGAUCCAUACUGCGGAAC GuuccGcAGuAuGGAucGGUsU 223 R-008380426-OOOH 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 270 R-008380426-OOOH 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUGGAUCGGUsU 271 R-008380427-OOOS 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 247 R-008380427-OOOS 2669 3 GCCGAUCCAUACUGCGGAA UUCCGCAGUAUGGAUCGGCUsU 267 R-008380428-OOOA 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 257 R-008380428-OOOA 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGuccAccAcUsU 259 R-008380429-000J 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 257 R-008380429-000J 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGUCCACCACUsU 256 R-008380430-OOOY 1665 2 GUGGUGGACUUCUCUCAAU B GuGGuGGAcuucucucAAuUsU B 233 R-008380430-OOOY 1665 2 GUGGUGGACUUCUCUCAAU AUUgagagaagUCCaCCaCUsU 251 R-008380431-OOOG 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 257 R-008380431-OOOG 1665 2 GUGGUGGACUUCUCUCAAU AuuGAGAGAAGuccAccAcUsU 234 R-008380432-OOOR 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 244 R-008380432-OOOR 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGUCCACCACGAUsU 240 R-008380433-OOOZ 1663 1 UCGUGGUGGACUUCUCUCA B ucGuGGuGGAcuucucucAUsU B 237 R-008380433-OOOZ 1663 1 UCGUGGUGGACUUCUCUCA UGAgagaagUCCaCCaCgaUsU 242 R-008380434-OOOH 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 261 R-008380434-OOOH 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUGGAUCGGUsU 221 R-008380435-OOOS 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 226
R-008380435-OOOS 2669 3 GCCGAUCCAUACUGCGGAA UUCcGcAGuAuGGAucGGcUsU 230 R-008380436-OOOA 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 244 R-008380436-OOOA 1663 1 UCGUGGUGGACUUCUCUCA UGAgagaagUCCaCCaCgaUsU 238 R-008380437-000J 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 244 R-008380437-000J 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGUCCACCACGAUsU 260 R-008380438-000T 1663 1 UCGUGGUGGACUUCUCUCA uGAGAGAAGuccAccAcGAUsU 243 R-008380438-OOOT 1663 1 UCGUGGUGGACUUCUCUCA B ucGuGGuGGAcuucucucAUsU B 237 R-008380441-OOOZ 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUaCUgCggAACUsU B 272 R-008380441-OOOZ 2670 4 CCGAUCCAUACUGCGGAAC GUUcCgcaGuAUggAuCgGUsU 273 R-008380444-000A 2670 4 CCGAUCCAUACUGCGGAAC B CCgAUCCaUaCUgcGgAaCUsU B 274 R-008380444-000A 2670 4 CCGAUCCAUACUGCGGAAC GuucCgCAgUAUGgAUCgGUsU 275 R-008380447-000B 2670 4 CCGAUCCAUACUGCGGAAC gUuCcggaGuau~gaUcGGUsU 276 R-008380447-OOOB 2670 4 CCGAUCCAUACUGCGGAAC B cCGA UCCauACUGCgGAACUsU B 277 R-008380450-OOOH 2669 3 GCCGAUCCAUACUGCGGAA uUccGCAGuaUgGaUCgGCUsU 278 R-008380450-OOOH 2669 3 GCCGAUCCAUACUGCGGAA B GCCgaUCCaUACUGCgGaAUsU B 279 R-008380453-000J 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGgACUUcUCUCAAUUsU B 280 R-008380453-000J 1665 2 GUGGUGGACUUCUCUCAAU AuugAgAgAAgUCcACCACUsU 281 R-008380456-000K 1665 2 GUGGUGGACUUCUCUCAAU alUgAGAgAgUCcACcaCUsU 282 R-008380456-OOOK 1665 2 GUGGUGGACUUCUCUCAAU B GUGgUgGaCUUCUCUCAAUUsU B 283 R-008380459-OOOL 1665 2 GUGGUGGACUUCUCUCAAUB GUGgUGgaCUUCUCUCaaUUsU B 284 R-008380459-OOOL 1665 2 GUGGUGGACUUCUCUCAAU AUugaGaGaAGu!cACCACUsU 285 R-008380462-OOOT 1663 1 UCGUGGUGGACUUCUCUCA B UCGGGUggaCuUCUCUCaUsU B 286 R-008380462-000T 1663 1 UCGUGGUGGACUUCUCUCA UGaGAGAaGUcCacCaCgAUsU 287 R-008380465-OOOU 2670 4 CCGAUCCAUACUGCGGAAC gUuccGCAgUaUgGAuCggUsU 288 R-008380465-OOOU 2670 4 CCGAUCCAUACUGCGGAAC B CCGaUCCAUaCuGC~gAAcUsU B 289 R-008380468-OOOV 2669 3 GCCGAUCCAUACUGCGGAA uUCcGcAgUAUGgAUcggCUsU 290 R-008380468-OOOV 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAuCCAUaCuGCgGaaUsU B 291 R-008380471-OOOB 2669 3 GCCGAUCCAUACUGCGGAA uUCcg~aGuAUggAUcGGCUsU 292 R-008380471-OOOB 2669 3 GCCGAUCCAUACUGCGGAA B gcCGAUCCAUACUGCGGAaUsU B 293 R-008380474-OOOC 2669 3 GCCGAUCCAUACUGCGGAA B GcCGAUCCAUACUGCGGaAUsU B 294 R-008380474-OOOC 2669 3 GCCGAUCCAUACUGCGGAA UUCcGCagUaggGAuCQGcUsU 295 R-008380477-OOOD 1665 2 GUGGUGGACUUCUCUCAAU B GUgGUGGaCUUCUCUcaaUUsU B 296 R-008380477-OOOD 1665 2 GUGGUGGACUUCUCUCAAU AuUQaGaGaAgUcCACcacUsU 297 R-008380480-000K 1665 2 GUGGUGGACUUCUCUCAAUaUuGAGAgaAgUccACCACUsU 298 R-008380480-OOOK 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUggA CUuCUCUCAA UUsU B 299 R-008380483-OOOL 1665 2 GUGGUGGACUUCUCUCAAUB gUGGUGGACUUCUcUCaAUUsU B 300 R-008380483-OOOL 1665 2 GUGGUGGACUUCUCUCAAU AUuGAGAGAAgUCCAcCaCUsU 301 R-008380486-OOOM 1663 1 UCGUGGUGGACUUCUCUCA B UCgUGGUGGaCUUCuCUCAUsU B 302 R-008380486-OOOM 1663 1 UCGUGGUGGACUUCUCUCA UGaGagAAguccAcCACgaUsU 303 R-008380489-OOON 1663 1 UCGUGGUGGACUUCUCUCA uGaGAGaAguCcacCAgA!UsU 304 R-008380489-OOON 1663 1 UCGUGGUGGACUUCUCUCA B uCGUggugGACUUCUCuCAUsU B 305 R-008380492-OOOV 1663 1 UCGUGGUGGACUUCUCUCA ugAGaGAaGUCcaCcaCGAUsU 306 R-008380492-OOOV 1663 1 UCGUGGUGGACUUCUCUCA B UCGuggUGgaCUUCucUCAUsU B 307 R-008380495-OOOW 2670 4 CCGAUCCAUACUGCGGAAC B CcGA UCCAUACUgcGgaACUsU B 308 R-008380495-000W 2670 4 CCGAUCCAUACUGCGGAAC GuuCcGCaGUAUGgauCGGUsU 309 R-008380498-OOOX 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUaCuGCGGAaCUsU B 310 R-008380498-OOOX 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUgGAUcgGUsU 311 R-008380501-OOOP 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAaCUsU B 312 R-008380501-OOOP 2670 4 CCGAUCCAUACUGCGGAAC GUUcCGCagUaugGAUcGgUsU 313 R-008380504-O0R 2670 4 CCGAUCCAUACUGCGGAAC gUUCcgCAGUaUGgAuCGGUsU 314 R-008380504-OOOR 2670 4 CCGAUCCAUACUGCGGAACB CCGaUCCAUACUGggaACUsUB 315
R-008380507-OOOS 2669 3 GCCGAUCCAUACUGCGGAAB GCCgaUCCAUACUgCGGaaUsUB 316 R-008380507-OOOS 2669 3 GCCGAUCCAUACUGCGGAA UCCgcAGUaUggauCGGcUsU 317 R-008380510-OOOY 1665 2 GUGGUGGACUUCUCUCAAU auUgAGAGAaGUccacCaCUsU 318 R-008380510-OOOY 1665 2 GUGGUGGACUUCUCUCAAU B GUggUggAcUUCuCuCaaUUsU B 319 R-008380513-OOOZ 1665 2 GUGGUGGACUUCUCUCAAUB gUggUggaCUUCUCUCaaUUsUB 320 R-008380513-OOOZ 1665 2 GUGGUGGACUUCUCUCAAU AUUAgAgaaGUccacCACUsU 321 R-008380516-OOOA 1665 2 GUGGUGGACUUCUCUCAAUB gUGUGGaCUUCUCUCAaUUsU B 322 R-008380516-OOOA 1665 2 GUGGUGGACUUCUCUCAAU AuuGAgaGAaGUCcACCaCUsU 323 R-008380519-OOOB 2670 4 CCGAUCCAUACUGCGGAAC B CgaUCCAuacUgcggaACUsU B 324 R-008380519-OOOB 2670 4 CCGAUCCAUACUGCGGAAC GUuCCGcAGUaUGgauCggUsU 325 R-008380522-OOOH 2669 3 GCCGAUCCAUACUGCGGAA B gCCgaUCCaUaCUgCggaaUsUB 326 R-008380522-OOOH 2669 3 GCCGAUCCAUACUGCGGAA UUcGCagUAuGGaUc~gCUsU 327 R-008380525-000J 2669 3 GCCGAUCCAUACUGCGGAA uUcCgCAGUAuGGaUC~gCUsU 328 R-008380525-000J 2669 3 GCCGAUCCAUACUGCGGAA B GccgaUCCaUACUGCgGAAUsU B 329 R-008380528-000K 2669 3 GCCGAUCCAUACUGCGGAA uucCgCAGuAUgGaUCGGCUsU 330 R-008380528-000K 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAuCCaUACUgCGGAAUsU B 331 R-008380531-OOOS 1665 2 GUGGUGGACUUCUCUCAAUalUlgagAGAAguCcACCAcUsU 332 R-008380531-OOOS 1665 2 GUGGUGGACUUCUCUCAAUB gUggUggAcuUCUCUCAaUUsU B 333 R-008380534-000T 1663 1 UCGUGGUGGACUUCUCUCA uGAGAgAAgUCCacCACGAUsU 334 R-008380534-OOOT 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGU!ggACuuCUCUCAUsU B 335 R-008380537-OOOU 1663 1 UCGUGGUGGACUUCUCUCA B UCgUgGUgGACUUCUCUCAUsU B 336 R-008380537-OOOU 1663 1 UCGUGGUGGACUUCUCUCA UGagAgAaGuCCAccaCGaUsU 337 R-008380540-OOOA 1663 1 UCGUGGUGGACUUCUCUCA B UCgugGUGACUUCUCUcAUsU B 338 R-008380540-OOOA 1663 1 UCGUGGUGGACUUCUCUCA UgAGAgAagUCCAcCACGAUsU 339 R-008380543-OOOB 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUcaUsUB 340 R-008380543-OOOB 1663 1 UCGUGGUGGACUUCUCUCA UgagagAAGucCACCAcgAUsU 341 R-008380546-OOOC 2670 4 CCGAUCCAUACUGCGGAAC gUuCcGCAgUAUggAUcgGUsU 342 R-008380546-OOOC 2670 4 CCGAUCCAUACUGCGGAAC B CCGaUCCAUACUGCGgaACUsUB 343 R-008380549-OOOD 2670 4 CCGAUCCAUACUGCGGAAC guUCCGcAGuAUggaUCGgsU 344 R-008380549-OOOD 2670 4 CCGAUCCAUACUGCGGAAC B CCgAUCcaUaCugCGGAACUsU B 345 R-008380552-OOOK 2669 3 GCCGAUCCAUACUGCGGAA B QcCgAUCCAUaCUgcGgaAUsUB 346 R-008380552-000K 2669 3 GCCGAUCCAUACUGCGGAA UuCCgCAGUAugGaUcGGCUsU 347 R-008380555-OOOL 2669 3 GCCGAUCCAUACUGCGGAA B GCCGaUCCAUACUGcGGAaUsU B 348 R-008380555-OOOL 2669 3 GCCGAUCCAUACUGCGGAA UucCgCAGUAuGgaUCgGcUsU 349 R-008380558-OOOM 1665 2 GUGGUGGACUUCUCUCAAUauuGAGaGAagUCCAcCACUsU 350 R-008380558-OOOM 1665 2 GUGGUGGACUUCUCUCAAU B GUgGUGGAcUUCUCuCAaUUsU B 351 R-008380561-OOOU 1665 2 GUGGUGGACUUCUCUCAAUB gUGgUGGACUUCUCUCAAUUsU B 352 R-008380561-OOOU 1665 2 GUGGUGGACUUCUCUCAAU AUuGaGAGAaGUCcacCACUsU 353 R-008380564-OOOV 1663 1 UCGUGGUGGACUUCUCUCA ugagaGAAGUcACcAga.sU 354 R-008380564-OOOV 1663 1 UCGUGGUGGACUUCUCUCA B UGUGGUGgaCUUCUCUCAUsU B 355 R-008380567-000W 2669 3 GCCGAUCCAUACUGCGGAA B GCcgaUCCAUaCUgCGGaAUsU B 356 R-008380567-000W 2669 3 GCCGAUCCAUACUGCGGAA UucCGCaGUaUGGauCGGCUsU 357 R-008380570-OOOC 1665 2 GUGGUGGACUUCUCUCAAU B GuGgUgGACUUcUCUcaAUU sU B 358 R-008380570-OOOC 1665 2 GUGGUGGACUUCUCUCAAU AUUgagAgAAGUcCAcCAcUsU 359 R-008380573-OOOD 1665 2 GUGGUGGACUUCUCUCAAU atUGAgAgaaGuCaCcAcUsU 360 R-008380573-OOOD 1665 2 GUGGUGGACUUCUCUCAAU B gUGguGgaCUuCUCUCAauUsU B 361 R-008380576-OOOE 1665 2 GUGGUGGACUUCUCUCAAU auUGagaGaAGUCcaCcAcUsU 362 R-008380576-OOOE 1665 2 GUGGUGGACUUCUCUCAAUB gUgGuggACUUCUCUCaAUUsU B 363 R-008380579-OOOF 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGgUggACUCUCUCAUsU B 364 R-008380579-OOOF 1663 1 UCGUGGUGGACUUCUCUCA UGAgAGaAguCCAcCACGaUsU 365 R-008380582-OOOM 1663 1 UCGUGGUGGACUUCUCUCA B UCgUggUGgACUUCUCUcaUsU B 366
R-008380582-OOOM 1663 1 UCGUGGUGGACUUCUCUCA UGAgagaAGUcCAcCAcGAUsU 367 R-008380585-OOON 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCaUaCUgCgGAaCUsU B 368 R-008380585-000N 2670 4 CCGAUCCAUACUGCGGAAC GuuCCgCAGUAuggAUCGgUsU 369 R-008380588-000P 2670 4 CCGAUCCAUACUGCGGAAC guUcCGCAguAuGgaUCGGUsU 370 R-008380588-000P 2670 4 CCGAUCCAUACUGCGGAAC B CCgAUcCAuALUGCgGaA CUsU B 371 R-008380591-000W 2670 4 CCGAUCCAUACUGCGGAAC gUUCcgCAGUAuggaUCgGUsU 372 R-008380591-OOOW 2670 4 CCGAUCCAUACUGCGGAAC B CcGaUCCAUaCUGCGGAaCUsU B 373 R-008380594-OOOX 2669 3 GCCGAUCCAUACUGCGGAA B gCCgAUCCAgaCUgCgGAAUsU B 374 R-008380594-OOOX 2669 3 GCCGAUCCAUACUGCGGAA UuCCGCAgUAUGGaUCGGcUsU 375 R-008380597-OOOY 2669 3 GCCGAUCCAUACUGCGGAA uuCCgcaggauGGaUCGGCUsU 376 R-008380597-OOOY 2669 3 GCCGAUCCAUACUGCGGAA BGCCGAUCCAUACUIgCGgaaUsU B 377 R-008380600-OOOR 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCcauACUgCGgaAUsU B 378 R-008380600-OOOR 2669 3 GCCGAUCCAUACUGCGGAA UUCCGcAGUaUGgaucGgCUsU 379 R-008380603-OOOS 1663 1 UCGUGGUGGACUUCUCUCA B UCgUggUggaCUUCUCUCaUsU B 380 R-008380603-OOOS 1663 1 UCGUGGUGGACUUCUCUCA UgaGAGAagUCcacCACgAUsU 381 R-008380606-000T 1663 1 UCGUGGUGGACUUCUCUCA uGAgAgAAguCCACCacGAUsU 382 R-008380606-OOOT 1663 1 UCGUGGUGGACUUCUCUCA B ucgUgGUgGaCUuCUCUCaUsU B 383 R-008380609-OOOU 2670 4 CCGAUCCAUACUGCGGAAC guUccGCaGUAUggAUCGgUsU 384 R-008380609-OOOU 2670 4 CCGAUCCAUACUGCGGAAC B CCgAUcCaUACUgCGgAA CUsU B 385 R-008380612-OOOA 2670 4 CCGAUCCAUACUGCGGAAC gUUCGCQagUAuGgaucGGUsU 386 R-008380612-OOOA 2670 4 CCGAUCCAUACUGCGGAAC B CCgAUCCAUACUGCGgaaCUsUB 387 R-008380615-OOOB 2670 4 CCGAUCCAUACUGCGGAAC gUUccGCAGUaUGgAUCGgUsU 388 R-008380615-OOOB 2670 4 CCGAUCCAUACUGCGGAAC B CCgaUCCaUaCUgCggaaCUsUB 389 R-008380618-OOOC 2670 4 CCGAUCCAUACUGCGGAAC B cCGaUCCaUACUGCggAACUsU B 390 R-008380618-OOOC 2670 4 CCGAUCCAUACUGCGGAAC GuUCcgCAGuaugGaU~gGUsU 391 R-008380621-000J 2669 3 GCCGAUCCAUACUGCGGAA B gCCGaUCCaUACUG~ggAaUsU B 392 R-008380621-000J 2669 3 GCCGAUCCAUACUGCGGAA UUcCgCAGUauGGAUcggCUsU 393 R-008380624-OOOK 2669 3 GCCGAUCCAUACUGCGGAA B gCCGaUCCaUaCUgCGgAAUsUB 394 R-008380624-000K 2669 3 GCCGAUCCAUACUGCGGAA UUCGCaGuAUGgaucgGCUsU 395 R-008380627-OOOL 2669 3 GCCGAUCCAUACUGCGGAA uUCCGCAGUAUGGaUcggcUsU 396 R-008380627-OOOL 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCgGAAUsU B 397 R-008380630-OOOT 2669 3 GCCGAUCCAUACUGCGGAA B GCCgAUCCAUACUGCGGAAUsU B 398 R-008380630-000T 2669 3 GCCGAUCCAUACUGCGGAA UUcCgCAGuAuggauCGGCUsU 399 R-008380633-OOOU 1665 2 GUGGUGGACUUCUCUCAAU B GugGUGgaC!UCUCUcAAUUsU B 400 R-008380633-OOOU 1665 2 GUGGUGGACUUCUCUCAAU AuUGAGaGaaGUCCAccaCUsU 401 R-008380636-OOOV 1665 2 GUGGUGGACUUCUCUCAAUB GUGGUGGACUUcUcUCaaUUsU B 402 R-008380636-OOOV 1665 2 GUGGUGGACUUCUCUCAAU AuUGAgAGAAgucCaCCACUsU 403 R-008380639-000W 1665 2 GUGGUGGACUUCUCUCAAU aULGAGAgaAguccACaCUsU 404 R-008380639-OOOW 1665 2 GUGGUGGACUUCUCUCAAUB GUGgUgGaCUUCUCUCAAUUsU B 405 R-008380642-OOOC 1663 1 UCGUGGUGGACUUCUCUCA B UCgUGgUgGACUUcUcUCaUsU B 406 R-008380642-OOOC 1663 1 UCGUGGUGGACUUCUCUCA gAGAgAAGuCCAcCaCGaUsU 407 R-008380645-OOOD 1663 1 UCGUGGUGGACUUCUCUCA B UCgUggUgGAcUUCUCUCAUsU B 408 R-008380645-OOOD 1663 1 UCGUGGUGGACUUCUCUCA UgaGAGaAGUCCAcCacGaUsU 409 R-008380648-OOOE 1663 1 UCGUGGUGGACUUCUCUCA ugAGaGaaGuCCAcCaCGAUsU 410 R-008380648-OOOE 1663 1 UCGUGGUGGACUUCUCUCA B UCGjgguGGaCUcUCUCaUsU B 411 R-008380651-OOOL 1663 1 UCGUGGUGGACUUCUCUCA B uCjuGGUgGaCUUCUCUcAUsU B 412 R-008380651-OOOL 1663 1 UCGUGGUGGACUUCUCUCA UgaGagaAGUcCacCAcGaUsU 413 R-008380654-OOOM 1663 1 UCGUGGUGGACUUCUCUCA uGagAGAAgucCACCACGaUsU 414 R-008380654-OOOM 1663 1 CGUGGUGGACUUCUCUCA B UCGUGgUGgACUuCuCUCAUsU B 415 R-008380655-000W 1665 2 UGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 250 R-008380655-000W 1665 2 GUGGUGGACUUCUCUCAAU AUuGAGAGAAgUCCAcCaCUsU 301
R-008380656-OOOE 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 244 R-008380656-OOOE 1663 1 UCGUGGUGGACUUCUCUCA UgAGAgAagUCCAcCACGAUsU 339 R-008380657-OOON 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 222 R-008380657-OOON 2670 4 CCGAUCCAUACUGCGGAAC GUUcCgcaGuAUggAuCgGUsU 273 R-008380658-OOOX 2670 4 CCGAUCCAUACUGCGGAAC gUuccGCAgUaUgGAuCggUsU 288 R-008380658-OOOX 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 261 R-008380659-OOOF 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 261 R-008380659-000F 2670 4 CCGAUCCAUACUGCGGAAC GUuCCGcAGUaUQgauCggUsU 325 R-008380660-000V 2669 3 GCCGAUCCAUACUGCGGAA uUccGCAGuaUgGaUCgGCUsU 278 R-008380660-OOOV 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 226 R-008380661-OOOD 1665 2 GUGGUGGACUUCUCUCAAU B GuGGuGGAcuucucucAAuUsU B 233 R-008380661-OOOD 1665 2 GUGGUGGACUUCUCUCAAU AuUGaGaGaAgUcCACcacUsU 297 R-008380662-000M 1663 1 UCGUGGUGGACUUCUCUCA B ucGuGGuGGAcuucucucAUsU B 237 R-008380662-000M 1663 1 UCGUGGUGGACUUCUCUCA UgaGAGAagUCcacCACgAUsU 381 R-008380663-000W 1663 1 UCGUGGUGGACUUCUCUCA B ucGuGGuGGAcuucucucAUsU B 237 R-008380663-000W 1663 1 UCGUGGUGGACUUCUCUCA UQAgAGaAguCCAcCACGaUsU 365 R-008380664-OOOE 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 244 R-008380664-OOOE 1663 1 UCGUGGUGGACUUCUCUCA UGaGagAAguccAcCACgaUsU 303 R-008380665-OOON 1663 1 UCGUGGUGGACUUCUCUCA ugAGaGaaGuCCAcCaCGAUsU 410 R-008380665-OOON 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 241 R-008380666-OOOX 1663 1 UCGUGGUGGACUUCUCUCA uGaGAGaAguCcacCAgA.sC 304 R-008380666-OOOX 1663 1 UCGUGGUGGACUUCUCUCA B ucGuGGuGGAcuucucucAUsU B 237 R-008380667-OOOF 1663 1 UCGUGGUGGACUUCUCUCA uGagAGAAgucCACCACGaUsU 414 R-008380667-OOOF 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 241 R-008380668-OOOP 2669 3 GCCGAUCCAUACUGCGGAA B GccGAuccAuAcuGcGGAAUsU B 224 R-008380668-OOOP 2669 3 GCCGAUCCAUACUGCGGAA UCCGCaGuAUGgaucgGCUsU 395 R-008380669-OOOY 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 226 R-008380669-OOOY 2669 3 GCCGAUCCAUACUGCGGAA UUCCgcAGUaUggauCGGcUsU 317 R-008380670-OOOM 1665 2 GUGGUGGACUUCUCUCAAU atTGAGAgaAguccACCaCUsU 404 R-008380670-OOOM 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 257 R-008380671-OOOW 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 241 R-008380671-000W 1663 1 UCGUGGUGGACUUCUCUCA UgagagAAGucCACCAcgAUsU 341 R-008380672-OOOE 1663 1 UCGUGGUGGACUUCUCUCA B ucGuGGuGGAcuucucucAUsU B 237 R-008380672-OOOE 1663 1 UCGUGGUGGACUUCUCUCA LgaGAGaAGUCCAcCacGaUsU 409 R-008380673-OOON 1663 1 UCGUGGUGGACUUCUCUCA uGAgAgAAguCCACCacGAUsU 382 R-008380673-OOON 1663 1 UCGUGGUGGACUUCUCUCA B ucGuGGuGGAcuucucucAUsU B 237 R-008380674-OOOX 2670 4 CCGAUCCAUACUGCGGAAC guUCCGcAGuAUggaUCGgsU 344 R-008380674-OOOX 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 222 R-008380675-OOOF 2669 3 GCCGAUCCAUACUGCGGAA uUcCgCAGUAuGGaUC~gCUsU 328 R-008380675-OOOF 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 247 R-008380676-OOOP 2669 3 GCCGAUCCAUACUGCGGAA uUCcg~aGuAUggAUcGGCUsU 292 R-008380676-OOOP 2669 3 GCCGAUCCAUACUGCGGAA B GccGAuccAuAcuGcGGAAUsU B 224 R-008380677-OOOY 2669 3 GCCGAUCCAUACUGCGGAA uucCgCAGuAUgGaUCGGCUsU 330 R-008380677-OOOY 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 226 R-008380678-OOOG 1665 2 GUGGUGGACUUCUCUCAAUauUgAGAGAaGUccacCaCUsU 318 R-008380678-OOOG 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 257 R-008380679-OOOR 1665 2 GUGGUGGACUUCUCUCAAU atUGAgAgaaGuCCaCcAcUsU 360 R-008380679-OOOR 1665 2 GUGGUGGACUUCUCUCAAU B GuGGuGGAcuucucucAAuUsU B 233 R-008380680-OOOE 1665 2 UGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 250 R-008380680-000E 1665 2 UGGUGGACUUCUCUCAAU AuugAgAgAAgUCcACCACUsU 281 R-008380681-OOON 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 222
R-008380681-OOON 2670 4 CCGAUCCAUACUGCGGAAC GuuCcGCaGUAUGgauCGGUsU 309 R-008380682-OOOX 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 244 R-008380682-OOOX 1663 1 UCGUGGUGGACUUCUCUCA UGaGAGAaGUcCacCaCgAUsU 287 R-008380683-OOOF 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 241 R-008380683-OOOF 1663 1 UCGUGGUGGACUUCUCUCA UgAGAgAAGuCCAcCaCGaUsU 407 R-008380684-OOOP 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 222 R-008380684-OOOP 2670 4 CCGAUCCAUACUGCGGAAC GuucCgCAgUAUGgAUCgGUsU 275 R-008380685-OOOY 2670 4 CCGAUCCAUACUGCGGAAC guUccGCaGUAUggAUCGgUsU 384 R-008380685-OOOY 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 261 R-008380686-OOOG 2669 3 GCCGAUCCAUACUGCGGAA B GccGAuccAuAcuGcGGAAUsU B 224 R-008380686-OOOG 2669 3 GCCGAUCCAUACUGCGGAA UucCGCaGUaUGGauCGGCUsU 357 R-008380687-OOOR 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 247 R-008380687-000R 2669 3 GCCGAUCCAUACUGCGGAA UuCCgCAGUAugGaUcGGCUsU 347 R-008380688-000Z 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 247 R-008380688-000Z 2669 3 GCCGAUCCAUACUGCGGAA UuCCGCAgUAUGGaUCGGcUsU 375 R-008380689-000H 2669 3 GCCGAUCCAUACUGCGGAA uuCCgcaggauGGaUCGGCUsU 376 R-008380689-000H 2669 3 GCCGAUCCAUACUGCGGAA B GccGAuccAuAcuGcGGAAUsU B 224 R-008380690-000X 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 247 R-008380690-000X 2669 3 GCCGAUCCAUACUGCGGAA UUCCjcAGUaUGgaucGgCUsU 379 R-008380691-000F 2670 4 CCGAUCCAUACUGCGGAAC B ccGAuccAuAcuGcGGAAcUsU B 220 R-008380691-OOOF 2670 4 CCGAUCCAUACUGCGGAAC GuuCCgCAGUAuggAUC gUsU 369 R-008380692-OOOP 1665 2 GUGGUGGACUUCUCUCAAUa!UlgagAGAAguCcACCAcUsU 332 R-008380692-OOOP 1665 2 GUGGUGGACUUCUCUCAAU B GuGGuGGAcuucucucAAuUsU B 233 R-008380693-OOOY 2670 4 CCGAUCCAUACUGCGGAAC B ccGAuccAuAcuGcGGAAcUsU B 220 R-008380693-OOOY 2670 4 CCGAUCCAUACUGCGGAAC GUUcCGCagUaugGAUcGgUsU 313 R-008380694-OOOG 2670 4 CCGAUCCAUACUGCGGAAC guUcCGCAguAuGgaUCGGUsU 370 R-008380694-OOOG 2670 4 CCGAUCCAUACUGCGGAAC B ccGAuccAuAcuGcGGAAcUsU B 220 R-008380695-OOOR 2670 4 CCGAUCCAUACUGCGGAAC gluCcgCaGuaugaUcGGUsU 276 R-008380695-OOOR 2670 4 CCGAUCCAUACUGCGGAAC B ccGAuccAuAcuGcGGAAcUsU B 220 R-008380696-OOOZ 2669 3 GCCGAUCCAUACUGCGGAA B GccGAuccAuAcuGcGGAAUsU B 224 R-008380696-OOOZ 2669 3 GCCGAUCCAUACUGCGGAA UUcGcQagUAuGGaUc~gQUsU 327 R-008380697-OOOH 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 226 R-008380697-OOOH 2669 3 GCCGAUCCAUACUGCGGAA UUcgCAGUauGGAUcggCUsU 393 R-008380698-OOOS 2669 3 GCCGAUCCAUACUGCGGAA B GccGAuccAuAcuGcGGAAUsU B 224 R-008380698-OOOS 2669 3 GCCGAUCCAUACUGCGGAA LucCgAGUAuGgaUCgGcUsU 349 R-008380699-OOOA 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 250 R-008380699-OOOA 1665 2 GUGGUGGACUUCUCUCAAU AUUgagAgAAGUcCAcCAcUsU 359 R-008380700-OOOA 1665 2 GUGGUGGACUUCUCUCAAU B GuGGuGGAcuucucucAAuUsU B 233 R-008380700-OOOA 1665 2 GUGGUGGACUUCUCUCAAUAAU gAgaaGUccacCACUsU 321 R-008380701-000J 1665 2 GUGGUGGACUUCUCUCAAUajUjgAGAgAgUcACcaCUsU 282 R-008380701-000J 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 257 R-008380702-000T 1663 1 UCGUGGUGGACUUCUCUCA uGAGAgAAgUCCacCACGAUsU 334 R-008380702-OOOT 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 244 R-008380703-OOOB 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 241 R-008380703-OOOB 1663 1 UCGUGGUGGACUUCUCUCA UGagAgAaGuCCAccaCGaUsU 337 R-008380704-OOOK 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 244 R-008380704-000K 1663 1 UCGUGGUGGACUUCUCUCA UgaGagaAGUcCacCAcGaUsU 413 R-008380705-OOOU 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 241 R-008380705-OOOU 1663 1 UCGUGGUGGACUUCUCUCA UGAgagaAGUcCAcCAcGAUsU 367 R-008380706-OOOC 1663 1 UCGUGGUGGACUUCUCUCA ugAGaGAaGUCcaCcaCGAUsU 306 R-008380706-OOOC 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUCAUsU B 244
R-008380707-OOOL 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 261 R-008380707-000L 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUgGAUcgGUsU 311 R-008380708-000V 2670 4 CCGAUCCAUACUGCGGAAC gUuCcGCAgUAUUggAUcgGUsU 342 R-008380708-000V 2670 4 CCGAUCCAUACUGCGGAAC B ccGAuccAuAcuGcGGAAcUsU B 220 R-008380709-000D 2670 4 CCGAUCCAUACUGCGGAAC gUUccGCAGUaUGgAUCGgUsU 388 R-008380709-000D 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 222 R-008380710-000T 2669 3 GCCGAUCCAUACUGCGGAA UUCC~cAgUAUGgAUcggCUsU 290 R-008380710-OOOT 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 247 R-008380711-OOOB 1665 2 GUGGUGGACUUCUCUCAAU auUGagaGaAGUCcaCcAcUsU 362 R-008380711-OOOB 1665 2 GUGGUGGACUUCUCUCAAU B GuGGuGGAcuucucucAAuUsU B 233 R-008380712-OOOK 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 257 R-008380712-000K 1665 2 GUGGUGGACUUCUCUCAAU AUugaGaGaAGuCcACCACUsU 285 R-008380713-OOOU 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 261 R-008380713-OOOU 2670 4 CCGAUCCAUACUGCGGAAC GuUCcgCAGuaugGaU~gGUsU 391 R-008380714-OOOC 2670 4 CCGAUCCAUACUGCGGAAC gUUCcgCAGUAuggaUCgGUsU 372 R-008380714-OOOC 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 261 R-008380715-OOOL 2669 3 GCCGAUCCAUACUGCGGAA uUCCGCAGUAUGGaUcggcUsU 396 R-008380715-OOOL 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 247 R-008380716-OOOV 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 250 R-008380716-OOOV 1665 2 GUGGUGGACUUCUCUCAAU AuUGAgAGAAgucCaCCACUsU 403 R-008380717-OOOD 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 257 R-008380717-OOOD 1665 2 GUGGUGGACUUCUCUCAAU AUuGaGAGAaGUCcacCACUsU 353 R-008380718-OOOM 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 250 R-008380718-OOOM 1665 2 GUGGUGGACUUCUCUCAAU AuuGAgaGAaGUCcACCaCUsU 323 R-008380719-000W 1665 2 GUGGUGGACUUCUCUCAAUaUuGAGAgaAgUccACCAUsU 298 R-008380719-000W 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 250 R-008380720-000K 1663 1 UCGUGGUGGACUUCUCUCA ugagaGAAG cACcACga.sU 354 R-008380720-OOOK 1663 1 UCGUGGUGGACUUCUCUCA B ucGuGGuGGAcuucucucAUsU B 237 R-008380721-OOOU 2670 4 CCGAUCCAUACUGCGGAAC gUU cgCAGUaUGgAuCGGUsU 314 R-008380721-OOOU 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAACUsU B 222 R-008380722-OOOC 2670 4 CCGAUCCAUACUGCGGAAC gUUCGCQagUAugaucGGUsU 386 R-008380722-OOOC 2670 4 CCGAUCCAUACUGCGGAAC B ccGAuccAuAcuGcGGAAcUsU B 220 R-008380723-OOOL 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 226 R-008380723-OOOL 2669 3 GCCGAUCCAUACUGCGGAA UUcCgCAGuAuggauCGGCUsU 399 R-008380724-OOOV 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCGGAAUsU B 226 R-008380724-OOOV 2669 3 GCCGAUCCAUACUGCGGAA UUCcGCagUaggGAuCGGcUsU 295 R-008380725-OOOD 1665 2 GUGGUGGACUUCUCUCAAU B GuGGuGGAcuucucucAAuUsU B 233 R-008380725-OOOD 1665 2 GUGGUGGACUUCUCUCAAU AuUGAGaGaaGUCCAccaCUsU 401 R-008380726-OOOM 1665 2 GUGGUGGACUUCUCUCAAUauuGAGaGAagUCCAcCACUsU 350 R-008380726-OOOM 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUCUCUCAAUUsU B 257 R-008380727-OOOW 2669 3 GCCGAUCCAUACUGCGGAA B gCCgAUCCAgaCUgCgGAAUsU B 374 R-008380727-000W 2669 3 GCCGAUCCAUACUGCGGAA UUCCgCagUaUggaUCggCUsU 227 R-008380728-OOOE 2669 3 GCCGAUCCAUACUGCGGAA uuccGcAGuAuGGAucGGcUsU 228 R-008380728-OOOE 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCcauACUgCQgaAUsU B 378 R-008380729-OOON 1665 2 GUGGUGGACUUCUCUCAAU B GUgGUGGAcUUCUCuCAaUUsU B 351 R-008380729-OOON 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGuccAccAcUsU 259 R-008380730-OOOC 1665 2 GUGGUGGACUUCUCUCAAU B gUGguGgaCUuCUCUCAauUsU B 361 R-008380730-OOOC 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGUCCACCACUsU 256 R-008380731-OOOL 1665 2 GUGGUGGACUUCUCUCAAUB gUGGUGGACUUCUcUCaAUUsU B 300 R-008380731-OOOL 1665 2 UGGUGGACUUCUCUCAAU uuGAGAGAAGuccAccAcUsU 234 R-008380732-OOOV 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUggACuuCUCUCAUsU B 335
R-008380732-OOOV 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGuccAccAcGAUsU 239 R-008380733-000D 1663 1 UCGUGGUGGACUUCUCUCA uGAGAGAAGuccAccAcGAUsU 243 R-008380733-000D 1663 1 UCGUGGUGGACUUCUCUCA B uCGUggugGACUUCUCuCAUsU B 305 R-008380734-OOOM 2670 4 CCGAUCCAUACUGCGGAAC B CCGaUCCAUaCuGCGgAAcUsU B 289 R-008380734-000M 2670 4 CCGAUCCAUACUGCGGAAC GUUccGcAGuAuGGAucGGUsU 246 R-008380735-000W 2670 4 CCGAUCCAUACUGCGGAAC B cCGA UCCauACUGCgGAACUsU B 277 R-008380735-000W 2670 4 CCGAUCCAUACUGCGGAAC GuuccGcAGuAuGGAucGGUsU 223 R-008380736-OOOE 2669 3 GCCGAUCCAUACUGCGGAA B gCCgaUCCaUaCUgCggaaUsUB 326 R-008380736-OOOE 2669 3 GCCGAUCCAUACUGCGGAA UUCCg!gaUggaUlggCUsg 225 R-008380737-OOON 2669 3 GCCGAUCCAUACUGCGGAA B gCCGaUCCaUaCUgCGgAAUsUB 394 R-008380737-OOON 2669 3 GCCGAUCCAUACUGCGGAA UUCCGCAGUAUGGAUCGGCUsU 267 R-008380738-OOOX 2669 3 GCCGAUCCAUACUGCGGAA uuccGcAGuAuGGAucGGcUsU 228 R-008380738-OOOX 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAuCCaUACUgCGGAAUsU B 331 R-008380739-OOOF 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGGACUUcUc UCaaUUsU B 402 R-008380739-OOOF 1665 2 GUGGUGGACUUCUCUCAAU AUUgagagaagUCCaCCaCUsU 263 R-008380740-OOOV 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUGgACUUcUCUCAAUUsU B 280 R-008380740-OOOV 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGUCCACCACUsU 256 R-008380741-OOOD 1663 1 UCGUGGUGGACUUCUCUCA B UCgUggUgGAcUUCUCUCAUsU B 408 R-008380741-OOOD 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGUCCACCACGAUsU 260 R-008380742-OOOM 1663 1 UCGUGGUGGACUUCUCUCA B ucgUgGUgGaCUuCUCUCaUsU B 383 R-008380742-OOOM 1663 1 UCGUGGUGGACUUCUCUCA UGAgagaagUCCaCCaCgaUsU 242 R-008380743-OOOW 2670 4 CCGAUCCAUACUGCGGAAC B CCGaUCCAUACUGCGgaACUsUB 343 R-008380743-OOOW 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUGGAUCGGUsU 254 R-008380744-OOOE 2670 4 CCGAUCCAUACUGCGGAAC B CCgAUCCaUaCUgcGgAaCUsU B 274 R-008380744-OOOE 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUGGAUCGGUsU 254 R-008380745-OOON 2670 4 CCGAUCCAUACUGCGGAAC B CCgaUCCAuacUgcggaACUsU B 324 R-008380745-OOON 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUGGAUCGGUsU 254 R-008380746-OOOX 2670 4 CCGAUCCAUACUGCGGAAC B CCgAUcCaUACUgCGgAACUsU B 385 R-008380746-OOOX 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUGGAUCGGUsU 221 R-008380747-OOOF 2670 4 CCGAUCCAUACUGCGGAAC B CcGaUCCAUaCUGCGGAaCUsU B 373 R-008380747-OOOF 2670 4 CCGAUCCAUACUGCGGAAC GuuccGcAGuAuGGAucGGUsU 223 R-008380748-OOOP 1665 2 GUGGUGGACUUCUCUCAAU B GuGgUgGACUUcUCUcaAUU sU B 358 R-008380748-OOOP 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGuccAccAcUsU 259 R-008380749-OOOY 1665 2 GUGGUGGACUUCUCUCAAUB gUggUggaCUUCUCUCaaUUsUB 320 R-008380749-OOOY 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGUCCACCACUsU 268 R-008380750-OOOM 1665 2 GUGGUGGACUUCUCUCAAU B GUGGUggA CUuCUCUCAA UUsU B 299 R-008380750-OOOM 1665 2 GUGGUGGACUUCUCUCAAU AUUgagagaagUCCaCCaCUsU 251 R-008380751-OOOW 1663 1 UCGUGGUGGACUUCUCUCA B UCgUGgUgGACUUcUcUCaUsU B 406 R-008380751-000W 1663 1 UCGUGGUGGACUUCUCUCA UGAgagaagUCCaCCaCgaUsU 238 R-008380752-OOOE 2670 4 CCGAUCCAUACUGCGGAAC B CcGA UCCAUACUgcGgaACUsU B 308 R-008380752-OOOE 2670 4 CCGAUCCAUACUGCGGAAC GUUC~gagUaUggaUlggUsU_ 266 R-008380753-OOON 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUACUGCGGAaCUsU B 312 R-008380753-OOON 2670 4 CCGAUCCAUACUGCGGAAC GUUccGcAGuAuGGAucGGUsU 246 R-008380754-OOOX 2670 4 CCGAUCCAUACUGCGGAAC B CCgAUcCAuACUGCgGaA CjsU B 371 R-008380754-OOOX 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUGGAUCGGUsU 221 R-008380755-OOOF 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAuCCAUaCuGCgGaaUsU B 291 R-008380755-OOOF 2669 3 GCCGAUCCAUACUGCGGAA UUCCgagUjagggaUlggCUsg 225 R-008380756-OOOP 2669 3 CCGAUCCAUACUGCGGAA B GCCgaUCCaUACUGCgGaAUsUB 279 R-008380756-OOOP 2669 3 CCGAUCCAUACUGCGGAA UUCCgCagUjaggaUlggCUsg 225 R-008380757-OOOY 2669 3 CCGAUCCAUACUGCGGAA B GCCgAUCCAUACUGCGGAAUsU B 398 R-008380757-OOOY 2669 3 GCCGAUCCAUACUGCGGAA UCCGCAGUAUGGAUCGGCUsU 248
R-008380758-OOOG 2669 3 GCCGAUCCAUACUGCGGAA uuccGcAGuAuGGAucGGcUsU 228 R-008380758-OOOG 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUgCGgaaUsUB 377 R-008380759-OOOR 1665 2 GUGGUGGACUUCUCUCAAU BGUGgUGgaCUUCUCUCaaUUsUB 284 R-008380759-OOOR 1665 2 GUGGUGGACUUCUCUCAAU AuuGAGAGAAGuccAccAcUsU 234 R-008380760-OOOE 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGGACUUCUCUcaUsUB 340 R-008380760-OOOE 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGUCCACCACGAUsU 240 R-008380761-000N 2670 4 CCGAUCCAUACUGCGGAAC B CCgaUCCaUaCUgCggaaCUsU B 389 R-008380761-OOON 2670 4 CCGAUCCAUACUGCGGAAC GUUCCgCagUaUggaUCggUsU 255 R-008380762-OOOX 2669 3 GCCGAUCCAUACUGCGGAA B GCcgaUCCAUaCUgCGGaAUsU B 356 R-008380762-OOOX 2669 3 GCCGAUCCAUACUGCGGAA UCcGcAGuAuGGAucGGcUsU 230 R-008380763-OOOF 1665 2 GUGGUGGACUUCUCUCAAU B gUGgUGGACUUCUCUCAAUUsU B 352 R-008380763-OOOF 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGUCCACCACUsU 256 R-008380764-000P 1665 2 GUGGUGGACUUCUCUCAAU B gUggUggAcuUCUCUCAaUUsU B 333 R-008380764-000P 1665 2 GUGGUGGACUUCUCUCAAU AUUgagagaagUCCaCCaCUsU 251 R-008380765-000Y 1665 2 GUGGUGGACUUCUCUCAAU B GUGgUgGaCUUCUCUCAAUUsU B 405 R-008380765-000Y 1665 2 GUGGUGGACUUCUCUCAAU AUUgagagaagUCCaCCaCUsU 251 R-008380766-000G 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGgUggACUCUCUCAUsU B 364 R-008380766-000G 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGuccAccAcGAUsU 239 R-008380767-000R 1663 1 UCGUGGUGGACUUCUCUCA uGAGAGAAGuccAccAcGAUsU 243 R-008380767-000R 1663 1 UCGUGGUGGACUUCUCUCA B UCGuggUGgaCUUCucUCAUsU B 307 R-008380768-000Z 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCaUaCUgCgGAaCUsUB 368 R-008380768-000Z 2670 4 CCGAUCCAUACUGCGGAAC GUUJ g~agUaUggaUCggUsU 266 R-008380769-000H 2669 3 GCCGAUCCAUACUGCGGAA B GCCGaUCCAUACUGcGGAaUsU B 348 R-008380769-000H 2669 3 GCCGAUCCAUACUGCGGAA UUCCGCAGUAUGGAUCGGCUsU 248 R-008380770-000X 2669 3 GCCGAUCCAUACUGCGGAA B GcgaUCCaUACUGCgGAAUsU B 329 R-008380770-000X 2669 3 GCCGAUCCAUACUGCGGAA UUCCGCAGUAUGGAUCGGCUsU 248 R-008380771-000F 1665 2 GUGGUGGACUUCUCUCAAUB gUGUGGaCUUCUCUCAaUUsU B 322 R-008380771-000F 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGUCCACCACUsU 268 R-008380772-OOOP 1663 1 UCGUGGUGGACUUCUCUCA B UCgUggUggaCUUCUCUCaUsU B 380 R-008380772-OOOP 1663 1 UCGUGGUGGACUUCUCUCA UGAgagaagUCCaC CgaUsU 238 R-008380773-OOOY 1663 1 UCGUGGUGGACUUCUCUCA B UCgUgGUgGACUUCUCUCAUsU B 336 R-008380773-OOOY 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGuccAccAcGAUsU 239 R-008380774-OOOG 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUGgaCUUCUCUCAUsU B 355 R-008380774-OOOG 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGUCCACCACGAUsU 240 R-008380775-OOOR 1663 1 UCGUGGUGGACUUCUCUCA B UCGjgguGGaCUcUCUCaUsU B 411 R-008380775-OOOR 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGUCCACCACGAUsU 260 R-008380776-OOOZ 1663 1 UCGUGGUGGACUUCUCUCA B UCgUggUGgACUUCUCUcaUsU B 366 R-008380776-OOOZ 1663 1 UCGUGGUGGACUUCUCUCA UGAgagaagUCCaCCaCgaUsU 242 R-008380777-OOOH 1663 1 UCGUGGUGGACUUCUCUCA uGAGAGAAGuccAccAcGAUsU 243 R-008380777-OOOH 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGgUGgACUuCuCUCAUsU B 415 R-008380778-OOOS 2670 4 CCGAUCCAUACUGCGGAAC B CCGaUCCAUACUG~ggaACUsU B 315 R-008380778-OOOS 2670 4 CCGAUCCAUACUGCGGAAC GUUCCGCAGUAUGGAUCGGUsU 221 R-008380779-OOOA 2670 4 CCGAUCCAUACUGCGGAAC B cCGaUCCaUACUGCggAACUsU B 390 R-008380779-OOOA 2670 4 CCGAUCCAUACUGCGGAAC GUUCCgCagUaUggaUCggUsU 255 R-008380780-OOOP 2669 3 GCCGAUCCAUACUGCGGAA B GcCgAUCCAUaCUgcGgaAUsUB 346 R-008380780-OOOP 2669 3 GCCGAUCCAUACUGCGGAA UUCcGcAGuAuGGAucGGcUsU 230 R-008380781-OOOY 2669 3 GCCGAUCCAUACUGCGGAABgcCGAUCCAUACUGCGGAaUsUB 293 R-008380781-OOOY 2669 3 GCCGAUCCAUACUGCGGAA UCCgCagUaUggaUCggCUsU 227 R-008380782-OOOG 2669 3 GCCGAUCCAUACUGCGGAA B GcCGAUCCAUACUGCGGaAUsU B 294 R-008380782-OOOG 2669 3 GCCGAUCCAUACUGCGGAA UCCgCagUaUggaUCggCUsU 227 R-008380783-OOOR 1665 2 GUGGUGGACUUCUCUCAAUIB GugGUGgaCLCUCUcAAUU sB 400
R-008380783-OOOR 1665 2 GUGGUGGACUUCUCUCAAU AUUgagagaagUCCaCCaCUsU 263 R-008380784-OOOZ 1665 2 GUGGUGGACUUCUCUCAAUB GUggUggACUUCuCuCaaUUsUB 319 R-008380784-OOOZ 1665 2 GUGGUGGACUUCUCUCAAU AUUgagagaagUCCaCCaCUsU 263 R-008380785-OOOH 1665 2 GUGGUGGACUUCUCUCAAU BGUGgUgGaCUUCUCUCAAUUsUB 283 R-008380785-OOOH 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGUCCACCACUsU 268 R-008380786-OOOS 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUaCuGCGGAaCUsU B 310 R-008380786-OOOS 2670 4 CCGAUCCAUACUGCGGAAC GtUCg ~agUaUggaUCggUsU_ 266 R-008380787-OOOA 2670 4 CCGAUCCAUACUGCGGAAC B CCGAUCCAUaCUgCggAACUsU B 272 R-008380787-OOOA 2670 4 CCGAUCCAUACUGCGGAAC GUUccGcAGuAuGGAucGGUsU 246 R-008380788-000J 2670 4 CCGAUCCAUACUGCGGAAC B CCgAUCCAUACUGCGgaaCUsU B 387 R-008380788-000J 2670 4 CCGAUCCAUACUGCGGAAC GUUCCgCagUaUggaUCggUsU 255 R-008380789-OOOT 2670 4 CCGAUCCAUACUGCGGAAC B CCgAUCcaUaCugCGGAACUsU B 345 R-008380789-000T 2670 4 CCGAUCCAUACUGCGGAAC GuuccGcAGuAuGGAucGGUsU 223 R-008380790-OOOG 2669 3 GCCGAUCCAUACUGCGGAA B gCCGaUCCaUACUG~ggAaUsU B 392 R-008380790-OOOG 2669 3 GCCGAUCCAUACUGCGGAA UCcGcAGuAuGGAucGGcUsU 230 R-008380791-OOOR 2669 3 GCCGAUCCAUACUGCGGAA B GCCGAUCCAUACUGCgGAAUsU B 397 R-008380791-OOOR 2669 3 GCCGAUCCAUACUGCGGAA UUCCGCAGUAUGGAUCGGCUsU 267 R-008380792-OOOZ 2669 3 GCCGAUCCAUACUGCGGAA B GCCgaUCCAUACUgCGGaaUsUB 316 R-008380792-OOOZ 2669 3 GCCGAUCCAUACUGCGGAA UUCCGCAGUAUGGAUCGGCUsU 267 R-008380793-OOOH 1665 2 GUGGUGGACUUCUCUCAAU B GUgGUGGaCUUCUCUcaaUUsU B 296 R-008380793-OOOH 1665 2 GUGGUGGACUUCUCUCAAU AUUGAGAGAAGuccAccAcUsU 259 R-008380794-OOOS 1665 2 GUGGUGGACUUCUCUCAAUB gUgGuggACUUCUCUCaAUUsU B 363 R-008380794-OOOS 1665 2 GUGGUGGACUUCUCUCAAU AuuGAGAGAAGuccAccAcUsU 234 R-008380795-OOOA 1663 1 UCGUGGUGGACUUCUCUCA B UCGUGGUggaCuUCUCUCaUsU B 286 R-008380795-OOOA 1663 1 UCGUGGUGGACUUCUCUCA UGAgagaagUCCaCCaCgaUsU 238 R-008380796-000J 1663 1 UCGUGGUGGACUUCUCUCA B UCgugGUGACUUCUCUcAUsU B 338 R-008380796-000J 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGUCCACCACGAUsU 240 R-008380797-OOOT 1663 1 UCGUGGUGGACUUCUCUCA B UCgUGGUGGaCUUCuCUCAUsU B 302 R-008380797-OOOT 1663 1 UCGUGGUGGACUUCUCUCA UGAGAGAAGUCCACCACGAUsU 260 R-008380798-OOOB 1663 1 UCGUGGUGGACUUCUCUCA B uCjuGGUgGaCUUCUCUcAUsU B 412 R-008380798-OOOB 1663 1 UCGUGGUGGACUUCUCUCA UGAgagaagUCCaCCaCgaUsU 242 R-008418316-OOOS 1662 43 CUCGUGGUGGACUUCUCUC B cucGuGGuGGAcuucucucUsU B 416 R-008418316-OOOS 1662 43 CUCGUGGUGGACUUCUCUC GAGAGAAGuccAccAcGAGUsU 417 R-008418319-OOOT 2046 44 CCUAUGGGAGUGGGCCUCA B ccuAuGGGAGuGGGccucAUsU B 418 R-008418319-000T 2046 44 CCUAUGGGAGUGGGCCUCA UGAGGcccAcucccAuAGGUsU 419 R-008418328-OOOB 568 45 AAGAACUCCCUCGCCUCGC BAAGAAcucccucGccucGcUsUB 420 R-008418328-OOOB 568 45 AAGAACUCCCUCGCCUCGC GCGAGGcGAGGGAGuucuuUsU 421 R-008418337-000K 2666 46 UCUGCCGAUCCAUACUGCG ucuGccGAuccAuAcuGcGUsU B 422 R-008418337-OOOK 2666 46 UCUGCCGAUCCAUACUGCG CGCAGuAuGGAucGGcAGAUsU 423 R-008418340-OOOS 1652 47 AGAGUCUAGACUCGUGGUGBAGAGucuAGAcucGuGGuGUsUB 424 R-008418340-OOOS 1652 47 AGAGUCUAGACUCGUGGUG CACcAcGAGucuAGAcucuUsU 425 R-008418343-OOOT 564 48 GAAGAAGAACUCCCUCGCC B GAAGAAGAAcucccucGccUsU B 426 R-008418343-000T 564 48 GAAGAAGAACUCCCUCGCC GGCGAGGGAGuucuucuucUsU 427 R-008418355-OOOC 52 49 UCAAGCCUCCAAGCUGUGC B ucAAGccuccAAGcuGuGcUsU B 428 R-008418355-OOOC 52 49 UCAAGCCUCCAAGCUGUGC GCAcAGcuuGGAGGcuuGAUsU 429 R-008418358-OOOD 55 50 AGCCUCCAAGCUGUGCCUU B AGccuccAAGcuGuGccuuUsU B 430 R-008418358-OOOD 55 50 AGCCUCCAAGCUGUGCCUU AAGGcAcAGcuuGGAGGcuUsU 431 R-008418361-OOOK 1659 51 AGACUCGUGGUGGACUUCU B AGAcucGuGGuGGAcuucuUsU B 432 R-008418361-000K 1659 51 AGACUCGUGGUGGACUUCU AGAAGuccAccAcGAGucuUsU 433 R-008418313-OOOR 564 48 GAAGAAGAACUCCCUCGCC B GAAGAAGAACUCCCUCGCCUsU B 434 R-008418313-OOOR 564 48 GAAGAAGAACUCCCUCGCC GGCGAGGGAGUUCUUCUUCUsU 435
R-008418322-000Z 1659 51 AGACUCGUGGUGGACUUCU B AGACUCGUGGUGGACUUCUsU B 436 R-008418322-000Z 1659 51 AGACUCGUGGUGGACUUCU AGAAGUCCACCACGAGUCUUsU 437 R-008418325-000A 2666 46 UCUGCCGAUCCAUACUGCG B UCUGCCGAUCCAUACUGCGUsU B 438 R-008418325-000A 2666 46 UCUGCCGAUCCAUACUGCG CGCAGUAUGGAUCGGCAGAUsU 439 R-008418331-000H 568 45 AAGAACUCCCUCGCCUCGC B AAGAACUCCCUCGCCUCGCUsU B 440 R-008418331-OOOH 568 45 AAGAACUCCCUCGCCUCGC GCGAGGCGAGGGAGUUCUUUsU 441 R-008418334-OOOJ 52 49 UCAAGCCUCCAAGCUGUGC B UCAAGCCUCCAAGCUGUGCUsU B 442 R-008418334-OOOJ 52 49 UCAAGCCUCCAAGCUGUGC GCACAGCUUGGAGGCUUGAUsU 443 R-008418346-OOOU 55 50 AGCCUCCAAGCUGUGCCUU B AGCCUCCAAGCUGUGCCUUUsU B 444 R-008418346-OOOU 55 50 AGCCUCCAAGCUGUGCCUU AAGGCACAGCUUGGAGGCUUsU 445 R-008418349-OOOV 1662 43 CUCGUGGUGGACUUCUCUC B CUCGUGGUGGACUUCUCUCUsU B 446 R-008418349-OOOV 1662 43 CUCGUGGUGGACUUCUCUC GAGAGAAGUCCACCACGAGUsU 447 R-008418352-OOOB 2046 44 CCUAUGGGAGUGGGCCUCA B CCUAUGGGAGUGGGCCUCAUsU B 448 R-008418352-OOOB 2046 44 CCUAUGGGAGUGGGCCUCA UGAGGCCCACUCCCAUAGGUsU 449 R-008418364-OOOL 1652 47 AGAGUCUAGACUCGUGGUG B AGAGUCUAGACUCGUGGUGUsUB 450 R-008418364-OOOL 1652 47 AGAGUCUAGACUCGUGGUG CACCACGAGUCUAGACUCUUsU 451
wherein: A, C, G, and U =ribose A, C, G or U a, g, c and u= 2'-deoxy-2'-fluoro A, G, C or U A, U, C and G= 2'-O-methyl (2'-OMe) A, U, C, or G A, U, C, and G =deoxy A, U, C, or G B = inverted abasic T= thymidine s = phosphorothioate linkage
FurtherSynthesis Stepsfor CommercialPreparation
[0334] Once analysis indicates that the target product purity has been achieved after the annealing step, the material is transferred to the tangential flow filtration (TFF) system for concentration and desalting, as opposed to doing this prior to the annealing step.
[0335] Ultrafiltration: The annealed product solution is concentrated using a TFF system containing an appropriate molecular weight cut-off membrane. Following concentration, the product solution is desalted via diafiltration using Milli-Q water until the conductivity of the filtrate is that of water.
[0336] Lyophilization: The concentrated solution is transferred to a bottle, flash frozen and attached to a lyophilizer. The product is then freeze-dried to a powder. The bottle is removed from the lyophilizer and is now ready for use.
Initial Screening Protocol (96-Well Plate Transfections)
Cell Culture Preparation:
[0337] Human hepatoma cell line, HepG2, or mouse hepatoma cell line, Hepal-6, were grown in modified Eagle's medium. All the culture media were supplemented with 10% fetal bovine serum, 100ptg/mL streptomycin, 1OOU/mL penicillin, and 1% sodium bicarbonate.
Transfection and Screening
[0338] A plasmid (pSiCheck-HepB-Partial, Ref ID :1104) was generated where a partial HBV genome was inserted in to the pSiCheck2 plasmid (Promega, Madison, WI) at the XhoI/NotI sites downstream of the Renilla Luciferase gene. The Renilla Luciferase mRNA transcript is then expressed upstream of the HBV transcript and therefore, siNAS targeted against HBV genome that cause knockdown of the HBV transcript will also result in knockdown of the Renilla Luciferase transcript. The level of Renilla Luciferase transcript can be determined by directly mestuing the amount of Renilla Luciferase activity. The plasmid also expresses the Firefly Luciferase gene that is unaffected by the HBV siNAs and can be used to normalize the data to take into account any variations in the amount of plasmid transfected into cells.
[0339] The partial HBV genome consisted of 2270 nts of the adw2 genotype A HBV virus (nts 158-2427 where EcoRI site is ntl).
Partial HBV DNA sequence (SEQ ID NO:503):
gagaacatcacatcaggatccctaggacccctgctcgtgttacaggcggggtttttcttgttgacaagaatcctcacaataccgcagagtctc gactcgtggtggacttctctcaattttctagggggatcacccgtgtgtcttggccaaaattcgcagtccccaacctccaatcactcaccaacct cctgtectccaatttgtectggttategetggatgtgtetgeggegttttatcatatteetettcatectgetgetatgcetcatettettattggtactt ctggattatcaaggtatgttgcccgtttgtcctctaattccaggaaccacaacaaccagtacgggaccctgcaaaacctgcacgactcctgct caaggcaactctatgtttccctcatgttgctgtacaaaacctacggatggaaattgcacctgtattcccatcccatcatcttgggctttcgcaaaa tacctatgggaatgggcctcagtccgtttctcatggctcagttcactagtgccatttgttcagtggttcgtagggctttcccccactgtttggcttt cagttatgtggatgatgtggtattgggggccaagtctgtacaacatcttgagtccctttataccgctattaccaattttcttttgtctttgggtataca tctaaaccctaacaaaacaaagagatggggttactccctaaacttcatgggatatgtaattggaagttggggaacattaccacaggatcatatt gtacaaaaacttaaacactgttttagaaaacttcctgttaataggcctattgattggaaagtatgtcaacgaattgtgggtcttttgggctttgccg ctccttttacacaatgtggataccctgccttaatgcctttgtatgcatgtatacaagctaaacaggcttttactttctcgccaacttacaaggccttt ctaagtaaacagtatatgaacctttaccccgttgcccggcaacggcctggtctgtgccaagtgtttgctgatgcaacccccactggctggggc ttggccattggccatcagcgcatgcgcggaacctttgcggctcctctgccgatccatactgcggaactcctagcagcttgttttgctcgcagc aggtctggagcaaaactcatcgggactgataattctgtcgtcctttctcggaaatatacatcttttccatggctgctaggttgtactgccaactgg attcttcgcgggacgtcctttgtttacgtcccgtcggcgctgaatcccgcggacgacccctctcggggtcgcttgggactctatcgtccccttc tccgtctgccgttccagccgaccacggggcgcacctctctttacgcggtctccccgtctgtgccttctcatctgccggaccgtgtgcacttcg cttcacctctgcacgttgcatggagaccaccgtgaacgcccatcagagcctgcccaaggtcttacataagaggactcttggactctcatcaat gtcaacgaccgaccttgaagcttacttcaaagactgtgtgtttaaagactgggaagagtcgggggaggaaattaggttaaaggtttatttatta ggaggctgtaggcataaattggtctgcgcaccagcaccatgcaactttttcacctctgcctaatcatctcttgtacatgtcccactgttcaagcct ccaagctgtgccttgggtggctttggggcatggacattgacccctataaagaatttggagctactgtggagttactctcatttttgccttctgactt ctttccttccgtccgggatctccttgataccgcctcagctctgtatcgggaagccttagagtctcctgagcattgctcacctcaccatacagcac tcaggcaagccattctctgctggggggaattaatgactctagctacctgggtgggtaataatttgcaagatccagcatccagggatctagtag tcaattatgttaatactaacatgggcctaaagatcaggcaactattgtggtttcatatttcttgtcttacttttggaagagatactgtgcttgagtattt ggtttctttcggagtgtggattcgcactcctcctgcctatagaccaccaaatgcccctatcttatcaacacttccggaaactactgttgttagacg aagggaccgaggtaggtcccctagaagaagaactccctcgcctcgcagacgaagatctcaatcgccgcgtcgcag
[0340] Cells were plated in all wells of tissue-culture treated, 96-well plates at a final count of 10000 cells/well in 100tL of the appropriate culture media. The cells were cultured for overnight after plating at 37°C in the presence of 5% Co 2 .
[0341] On the next day, complexes containing siNA, pSiCheck-HepB-Partial plasmid, and Lipofectamine (Invitrogen) were created as follows. A solution of siNA and plasmid was generated containing 500nM siNA and 30ng/ul plasmid. In parallel, a solution of Lipofectamine diluted 33-fold in OPTI-MEM was prepared. After incubation of RNAiMax/OPTI-MEM solution at room temperature for 5 min, an equal volume of the siNA solution and the RNAiMax solution were added together for each of the siNAs.
[0342] Mixing resulted in a solution of siNA/RNAiMax where the concentration of siNA was 60 nM and the plasmid was 3.6ng/pl. This solution was incubated at room temperature for minutes. After incubation, 20 L of the solution was added to each of the relevant wells. The final concentration of siNA in each well was 10 nM and 0.6ng/pl for the plasmid. The final volume of Lipofectamine in each well was 0.3ul.
[0343] For 12-point dose response curve studies, the siNA series are 6-fold serial dilution starting at 30nM or 4-fold serial dilution starting at 40nM. All transfections were set up as multiple biological replicates.
[0344] The time of incubation with the Lipofectamine-siNA complexes was 48 hours and there was no change in media between transfection and harvesting for screening and dose response curve studies.
Luciferase Assays
[0345] 50pl of the Dual-GloTM Firefly reagent (Promega, Madison, WI) was added to each well of the 96 well-plate and then the plate was incubated at room temperature for 10 minutes. The firefly luminescence was then read on a Wallac Envison plate reader using the Luciferase protocol. 50pl of a 1:100 dilution of the Stop & Glo substrate (Promega, Madison, WI) was added to each well of the 96 well-plate and then the plate was incubated at room temperature for minutes. The Renilla luminescence was then read on a Wallac Envison plate reader using the Luciferase protocol.
Calculations
[0346] The expression level of the gene of interest and % inhibition of gene expression (%KD) was calculated using a Comparative method:
L_firefly = log 2 (firefly luc luminosity) L_renilla = log 2 (renilla luc luminosity)
dL = Lfirefly - L renilla ddL(log2(foldchange)) = dL_siRNA - dL_NTC
Relative expression level = 2-ddL
% KD = 100 x (1 - 2-d)
[0347] The non-targeting control siNA was, unless otherwise indicated, chosen as the value against which to calculate the percent inhibition (knockdown) of gene expression, because it is the most relevant control.
[0348] Additionally, only normalized data, which reflects the general health of the cell and quality of the plasmid and siNAs, was examined. This was done by looking at the level of two different mRNAs in the treated cells, the first being the target mRNA and the second being the normalizer mRNA. This is done by comparing the log2 luminescence for renilla luciferase (target) in each well relative to the log2 luminescence forfirefly luciferase (normalizer) for the entire plate. This allowed for elimination of siNAs that might be potentially toxic to cells rather than solely knocking down the gene of interest.
[0349] All calculations of IC5 0s were performed using R.2.9.2 software. The data were analyzed using the sigmoidal dose-response (variable slope) equation for simple ligand binding. In all of the calculations of the percent inhibition (knock-down), the calculation was made relative to the normalized level of expression of the gene of interest in the samples treated with the non-targeting control (Ctrl siNA) unless otherwise indicated.
Results:
[0350] The HBV siNAs were designed and synthesized as described previously. Various siNAs were screened in HepG2 or Hepal-6 cells transfected with the pSiCheck-HepB-Partial plasmid. The log 2(fold change) in HBV mRNA expression levels upon treatment with various modified HBV siNAs in human cells is shown in Table 3a and b. Each screen was performed at 48 hrs using the luicferase assay method.
Table 3a: Primary screening data in HepG2 Cells transfected with pSiCheck-HepB-partial (n= 2), recorded as log 2(fold change) in HBV mRNA expression levels
siNA Duplex ID Mean log 2(fold SD log 2(fold change) change) R-008351268-OOOC 6.91 0.06 R-008351278-000V 5.40 0.02 R-008351342-OOOA 5.18 0.16 R-008351172-OOOH 5.43 0.09 R-008351362-OOOK 4.33 0.09 R-008351389-OOOF 4.82 0.21 R-008351306-OOOR 5.12 0.08 R-008351372-OOOC 4.33 0.22 R-008351317-000S 5.07 0.06 R-008351210-OOOW 4.79 0.08 R-008351329-OOOB 4.71 0.20 R-008351350-OOOA 4.87 0.20 R-008351303-OOOP 4.97 0.06 R-008351229-OOOS 4.99 0.06 R-008351386-OOOE 5.08 0.08 R-008351300-OOON 4.61 0.24 R-008351263-OOOJ 4.49 0.02 R-008351190-OOOA 4.62 0.04 R-008351326-OOOA 4.67 0.09 R-008351260-OOOH 4.44 0.16 R-008351195-OOOU 4.37 0.08
R-008351323-OOOZ 4.42 0.21 R-008351367-OOOD 4.11 0.06 R-008351251-OOOZ 4.78 0.14 R-008351240-OOOY 4.51 0.28 R-008351178-OOOK 4.26 0.17 R-008351353-OOOB 4.43 0.00 R-008351297-OOOW 4.13 0.12 R-008351175-OOOJ 1.36 0.20 R-008351181-OOOS 2.68 0.12 R-008351184-OOOT 2.45 0.02 R-008351187-OOOU 0.51 0.13 R-008351192-OOOT 0.39 0.05 R-008351198-OOOV 0.52 0.22 R-008351201-OOOM 2.18 0.27 R-008351204-OOON 0.48 0.25 R-008351207-OOOP 0.90 0.02 R-008351212-OOON 0.45 0.26 R-008351215-OOOP 2.94 0.12 R-008351218-OOOR 1.24 0.02 R-008351220-OOON 0.33 0.09 R-008351223-OOOP 0.64 0.17 R-008351226-OOOR 0.22 0.16 R-008351232-OOOY 2.98 0.32 R-008351235-OOOZ 0.85 0.01 R-008351237-OOOS 3.12 0.11 R-008351243-OOOZ 3.79 0.09 R-008351246-OOOA 0.35 0.20 R-008351248-OOOT 0.52 0.29 R-008351254-OOOA 3.87 0.30 R-008351257-OOOB 3.13 0.05 R-008351266-OOOK 3.84 0.13 R-008351271-OOOJ 3.04 0.18 R-008351274-OOOK 0.33 0.24 R-008351276-OOOC 2.09 0.15 R-008351281-OOOB 3.71 0.20 R-008351284-OOOC 3.96 0.16 R-008351287-OOOD 2.63 0.06 R-008351290-OOOK 0.47 0.27 R-008351292-OOOC 4.11 0.17 R-008351294-OOOV 4.13 0.16 R-008351309-OOOS 1.91 0.16 R-008351312-OOOY 1.32 0.05 R-008351315-OOOZ 2.25 0.05 R-008351320-OOOY 0.48 0.22 R-008351332-OOOH 3.23 0.01
R-008351334-OOOA 2.09 0.60 R-008351337-OOOB 3.65 0.15 R-008351339-OOOU 0.11 0.27 R-008351345-OOOB 2.93 0.01 R-008351348-OOOC 0.95 0.24 R-008351356-000C 1.54 0.49 R-008351359-OOOD 0.88 0.15 R-008351364-OOOC 4.01 0.10 R-008351370-OOOK 4.17 0.04 R-008351374-OOOV 1.59 0.16 R-008351377-OOOW 1.37 0.22 R-008351380-OOOC 1.80 0.08 R-008351383-OOOD 0.79 0.13 R-008351392-OOOM 3.91 0.01 R-008351395-OOON 4.03 0.05 R-008351398-OOOP 0.69 0.12 R-008351401-OOOG 1.03 0.19 R-008351404-OOOH 2.57 0.10
Table 3b: Primary screening data in HepG2 Cells transfected with pSiCheck-HepB-partial (n= 2), recorded as log 2(fold change) in HBV mRNA expression levels
siNA Duplex ID Mean log 2(fold SD log 2(fold change) change) R-008351268-OOOC 6.41 0.01 R-008351278-OOOV 5.26 0.05 R-008351342-OOOA 5.01 0.13 R-008351172-OOOH 5.29 0.01 R-008351362-OOOK 4.50 0.20 R-008351389-OOOF 4.99 0.05 R-008351306-OOOR 4.81 0.10 R-008351372-OOOC 4.85 0.01 R-008351317-OOOS 5.45 0.04 R-008351210-OOOW 4.71 0.17 R-008351329-OOOB 4.69 0.16 R-008351350-OOOA 4.79 0.00 R-008351303-OOOP 4.57 0.04 R-008351229-OOOS 4.87 0.23 R-008351386-OOOE 4.98 0.28 R-008351300-OOON 4.48 0.06 R-008351263-OOOJ 4.96 0.08 R-008351190-OOOA 4.61 0.04
R-008351326-OOOA 4.80 0.07 R-008351260-OOOH 4.16 0.18 R-008351195-OOOU 4.35 0.00 R-008351323-OOOZ 4.07 0.06 R-008351367-OOOD 4.28 0.08 R-008351251-OOOZ 4.96 0.91 R-008351240-OOOY 4.63 0.01 R-008351178-OOOK 4.07 0.12 R-008351353-OOOB 4.20 0.02 R-008351297-OOOW 4.10 0.21 R-008351175-OOOJ 1.26 0.28 R-008351181-OOOS 2.56 0.03 R-008351184-OOOT 2.57 0.25 R-008351187-OOOU 0.52 0.01 R-008351192-OOOT 0.01 0.25 R-008351198-OOOV 0.49 0.18 R-008351201-OOOM 2.26 0.11 R-008351204-OOON 0.74 0.19 R-008351207-OOOP 0.58 0.34 R-008351212-OOON 0.18 0.26 R-008351215-OOOP 3.17 0.14 R-008351218-OOOR 1.32 0.04 R-008351220-OOON 0.15 0.05 R-008351223-OOOP 0.42 0.05 R-008351226-OOOR 0.49 0.07 R-008351232-OOOY 3.06 0.27 R-008351235-OOOZ 0.93 0.09 R-008351237-OOOS 2.68 0.01 R-008351243-OOOZ 3.85 0.10 R-008351246-OOOA 0.34 0.01 R-008351248-OOOT 0.93 0.37 R-008351254-OOOA 4.15 0.17 R-008351257-OOOB 3.61 0.00 R-008351266-OOOK 3.87 0.25 R-008351271-OOOJ 2.67 0.26 R-008351274-OOOK 0.43 0.16 R-008351276-OOOC 1.85 0.15 R-008351281-OOOB 3.76 0.97 R-008351284-OOOC 4.29 0.00 R-008351287-OOOD 2.06 0.09 R-008351290-OOOK 0.11 0.14 R-008351292-OOOC 3.99 0.01 R-008351294-OOOV 4.07 0.18 R-008351309-OOOS 1.99 0.21 R-008351312-OOOY 1.11 0.09
R-008351315-OOOZ 2.15 0.06 R-008351320-OOOY 0.35 0.19 R-008351332-OOOH 2.86 0.04 R-008351334-OOOA 1.71 0.01 R-008351337-OOOB 3.59 0.01 R-008351339-OOOU 0.21 0.18 R-008351345-OOOB 3.09 0.23 R-008351348-OOOC 0.92 0.12 R-008351356-OOOC 1.60 0.09 R-008351359-OOOD 0.76 0.01 R-008351364-OOOC 4.37 0.05 R-008351370-OOOK 3.99 0.19 R-008351374-OOOV 1.06 0.10 R-008351377-OOOW 1.20 0.09 R-008351380-OOOC 1.33 0.25 R-008351383-OOOD 0.24 0.19 R-008351392-OOOM 3.83 0.08 R-008351395-OOON 3.58 0.01 R-008351398-OOOP 0.53 0.01 R-008351401-OOOG 1.04 0.09 R-008351404-OOOH 2.82 0.04
[0351] A subset of siNAs from Table 3a and Table 3b having a large log2(fold change) in the primary screen were re-racked onto a single 96-well plate in triplicate and screened again in human HepG2 cells transfected with pSiCheckHepB-partial in order to confirm knockdown and to directly compare siNAs that were synthesized on different plates. Table 4 summarizes the data. The siNAs were ranked based on log2(fold change) in HBV mRNA expression levels.
Table 4. Log2(fold change) of various siNAs tested on a re-racked plate
siNA Duplex ID Mean log 2(fold SD log 2(fold change) change) R-008351268-OOOC 6.38 0.17 R-008351278-OOOV 5.71 0.23 R-008351342-OOOA 5.21 0.22 R-008351172-OOOH 5.00 0.11 R-008351362-OOOK 4.58 0.20 R-008351389-OOOF 4.84 0.44 R-008351306-OOOR 5.13 0.18 R-008351372-OOOC 4.77 0.09 R-008351317-OOOS 5.19 0.13 R-008351210-OOOW 4.68 0.09
R-008351329-OOOB 4.71 0.08 R-008351350-OOOA 4.54 0.16 R-008351303-OOOP 4.94 0.05 R-008351229-OOOS 4.85 0.13 R-008351386-OOOE 5.03 0.11 R-008351300-OOON 4.51 0.20 R-008351263-OOOJ 4.74 0.28 R-008351190-OOOA 4.50 0.15 R-008351326-OOOA 4.47 0.05 R-008351260-OOOH 4.33 0.27 R-008351195-OOOU 4.30 0.03 R-008351323-OOOZ 4.30 0.08 R-008351367-OOOD 4.23 0.06 R-008351251-OOOZ 4.21 0.09 R-008351240-OOOY 4.14 0.22 R-008351178-OOOK 4.11 0.08 R-008351353-OOOB 3.86 0.06 R-008351297-OOOW 3.79 0.20
[0352] Select high ranking (larger log 2(fold change) value) siNAs from Table 4 were further analyzed for efficacy and potency in HepG2cells transfected with pSiCheck-HepB-partial Plasmid. Cells were treated with a 1:6 serial dose titration of siNA in order to establish a value. The Potency50 is the calculated siNA transfection concentration that produces 50% target mRNA knockdown. The results for these siNAs are shown in Table 5.
Table 5: Dose response data for various siNAs in HepG2 -Human hepatoma cell line transfected with pSiCheck-HepB-partial. Calculated maximum log 2(fold change) is determined from the dose response curve. siNA Duplex ID ssDRC max log Potency50 2(fold change) (nM) R-008351268-OOOC 7.00 0.02 R-008351278-OOOV 5.59 0.11 R-008351342-OOOA 5.59 0.05 R-008351172-OOOH 5.52 0.07 R-008351362-OOOK 5.52 0.15 R-008351389-OOOF 5.33 0.16 R-008351306-OOOR 5.25 0.19 R-008351372-OOOC 5.21 0.09 R-008351317-OOOS 5.12 0.11 R-008351210-OOOW 4.93 0.17 R-008351329-OOOB 4.92 0.05 R-008351350-OOOA 4.92 0.18
R-008351303-000P 4.81 0.15 R-008351229-000S 4.81 0.03 R-008351386-000E 4.71 0.34 R-008351300-000N 4.63 0.15 R-008351263-000J 4.46 0.16 R-008351190-000A 4.45 0.10
[0353] Select top performing sequences idenitified in the previous screens, based on high ranking (larger log 2(fold change) value and low Potency 5 values from Table 5 were then syntheisezd in various chemical modifications on both passenger and guide strand. These siNAs were then analyzed for efficacy and potency in Hepal-6 cells transfected with pSiCheck-HepB partial Plasmid. The log 2(fold change) in HBV mRNA expression levels upon treatment with various modified HBV siNAs in human cells is shown in Table 6A. Each screen was performed at 48 hrs using the luicferase assay method.
Table 6A. Primary screening data of Lead Optimized HBV Sequences in Hepal-6 Cells transfected with pSiCheck-HepB-partial (n = 2), recorded as log 2(fold change) in HBV mRNA expression levels
Mean log 2(fold SD log 2(fold siNA Duplex ID change) change) R-008380633-OOOU 6.29 0.03 R-008380783-OOOR 6.23 0.11 R-008380330-OOON 6.19 0.06 R-008380665-OOON 6.18 0.15 R-008380648-OOOE 6.15 0.20 R-008380558-OOOM 6.08 0.10 R-008380365-OOOJ 5.96 0.20 R-008380406-OOOY 5.96 0.10 R-008380764-OOOP 5.95 0.02 R-008380394-OOOC 5.94 0.21 R-008380740-OOOV 5.93 0.03 R-008380408-OOOR 5.90 0.11 R-008380576-OOOE 5.87 0.02 R-008380645-OOOD 5.86 0.03
R-008380777-OOOH 5.85 0.18 R-008380730-OOOC 5.82 0.10 R-008380363-OOOS 5.81 0.07 R-008380767-OOOR 5.79 0.23 R-008380359-OOOB 5.79 0.09 R-008380763-OOOF 5.76 0.04 R-008380362-OOOH 5.75 0.19 R-008380771-OOOF 5.72 0.09 R-008380340-OOOF 5.72 0.05 R-008380417-OOOZ 5.66 0.09 R-008380377-OOOU 5.65 0.07 R-008380389-OOOD 5.63 0.19 R-008380772-OOOP 5.58 0.03 R-008380785-OOOH 5.56 0.03 R-008380680-OOOE 5.47 0.02 R-008380453-OOOJ 5.46 0.02 R-008380750-OOOM 5.42 0.02 R-008380776-OOOZ 5.41 0.30 R-008380391-OOOB 5.40 0.07 R-008380793-OOOH 5.35 0.09 R-008380402-OOON 5.33 0.26 R-008380516-OOOA 5.27 0.03 R-008380492-OOOV 5.22 0.25 R-008380718-OOOM 5.21 0.02 R-008380603-OOOS 5.20 0.04 R-008380729-OOON 5.19 0.02 R-008380795-OOOA 5.14 0.09 R-008380429-OOOJ 5.13 0.07 R-008380760-OOOE 5.12 0.04 R-008380459-OOOL 5.12 0.11 R-008380733-OOOD 5.06 0.22 R-008380683-OOOF 5.04 0.04 R-008380564-OOOV 5.02 0.01 R-008380582-OOOM 5.01 0.15 R-008380418-OOOH 4.98 0.04 R-008380540-OOOA 4.93 0.07 R-008380651-OOOL 4.91 0.12
R-008380774-OOOG 4.89 0.01 R-008380489-OOON 4.86 0.14 R-008380656-OOOE 4.86 0.11 R-008380739-OOOF 4.79 0.01 R-008380462-OOOT 4.73 0.08 R-008380324-OOOF 4.72 0.06 R-008380773-OOOY 4.69 0.17 R-008380361-OOOZ 4.69 0.35 R-008380798-OOOB 4.69 0.10 R-008380432-OOOR 4.66 0.16 R-008380796-OOOJ 4.63 0.03 R-008380794-OOOS 4.63 0.03 R-008380420-OOOF 4.61 0.11 R-008380543-OOOB 4.59 0.16 R-008380642-OOOC 4.57 0.04 R-008380731-OOOL 4.55 0.11 R-008380346-OOOH 4.50 0.02 R-008380378-OOOC 4.47 0.29 R-008380477-OOOD 4.46 0.01 R-008380405-OOOP 4.43 0.16 R-008380749-OOOY 4.42 0.27 R-008380537-OOOU 4.41 0.12 R-008380436-OOOA 4.40 0.19 R-008380742-OOOM 4.34 0.13 R-008380765-OOOY 4.31 0.09 R-008380353-OOOZ 4.29 0.10 R-008380712-OOOK 4.22 0.02 R-008380784-OOOZ 4.20 0.07 R-008380379-OOOL 4.20 0.24 R-008380753-OOON 4.19 0.09 R-008380689-OOOH 4.15 0.06 R-008380654-OOOM 4.15 0.14 R-008380374-OOOT 4.13 0.32 R-008380782-OOOG 4.08 0.07 R-008380480-OOOK 4.06 0.15 R-008380766-OOOG 4.02 0.26 R-008380348-OOOA 4.01 0.21
R-008380734-OOOM 3.97 0.09 R-008380486-OOOM 3.97 0.09 R-008380579-OOOF 3.97 0.11 R-008380437-OOOJ 3.93 0.14 R-008380781-OOOY 3.91 0.10 R-008380416-OOOR 3.88 0.29 R-008380343-OOOG 3.88 0.19 R-008380704-OOOK 3.88 0.29 R-008380797-OOOT 3.85 0.13 R-008380736-OOOE 3.84 0.02 R-008380706-OOOC 3.83 0.20 R-008380751-OOOW 3.83 0.17 R-008380756-OOOP 3.83 0.21 R-008380732-OOOV 3.79 0.20 R-008380321-OOOE 3.75 0.05 R-008380755-OOOF 3.71 0.27 R-008380528-OOOK 3.67 0.08 R-008380531-OOOS 3.64 0.20 R-008380426-OOOH 3.60 0.02 R-0083 80311 -OOOM 3.58 0.18 R-008380737-OOON 3.54 0.20 R-008380757-OOOY 3.52 0.07 R-008380787-OOOA 3.46 0.08 R-008380726-OOOM 3.46 0.35 R-008380678-OOOG 3.44 0.18 R-008380597-OOOY 3.40 0.09 R-008380686-OOOG 3.37 0.11 R-008380636-OOOV 3.31 0.05 R-008380791-OOOR 3.27 0.24 R-008380567-OOOW 3.27 0.08 R-008380624-OOOK 3.21 0.02 R-008380775-OOOR 3.21 0.04 R-008380315-OOOX 3.19 0.04 R-008380407-OOOG 3.13 0.21 R-008380741-OOOD 3.10 0.20 R-008380352-OOOR 3.09 0.07 R-008380450-OOOH 3.08 0.09
R-008380549-OOOD 3.05 0.08 R-008380789-OOOT 3.04 0.11 R-008380746-OOOX 3.03 0.10 R-008380510-OOOY 3.00 0.04 R-008380703-OOOB 3.00 0.02 R-008380664-OOOE 2.98 0.15 R-008380754-OOOX 2.98 0.06 R-008380759-OOOR 2.96 0.14 R-008380399-OOOW 2.94 0.13 R-008380735-OOOW 2.92 0.13 R-008380609-OOOU 2.86 0.16 R-008380386-OOOC 2.85 0.16 R-008380778-OOOS 2.82 0.11 R-008380428-OOOA 2.78 0.31 R-008380483-OOOL 2.75 0.07 R-008380444-OOOA 2.73 0.13 R-008380716-OOOV 2.69 0.16 R-008380356-OOOA 2.69 0.06 R-008380667-OOOF 2.69 0.23 R-008380671-OOOW 2.68 0.10 R-008380705-OOOU 2.67 0.16 R-008380410-OOON 2.65 0.16 R-008380431-OOOG 2.61 0.07 R-008380738-OOOX 2.60 0.06 R-008380427-OOOS 2.54 0.13 R-008380313-OOOE 2.49 0.30 R-008380728-OOOE 2.43 0.05 R-008380792-OOOZ 2.40 0.02 R-008380588-OOOP 2.40 0.17 R-008380397-OOOD 2.39 0.01 R-008380423-OOOG 2.37 0.07 R-008380434-OOOH 2.37 0.14 R-008380316-OOOF 2.27 0.21 R-008380555-OOOL 2.25 0.00 R-008380790-OOOG 2.23 0.09 R-008380398-OOOM 2.21 0.07 R-008380699-OOOA 2.16 0.25
R-008380719-OOOW 2.15 0.22 R-008380335-OOOG 2.14 0.45 R-008380618-OOOC 2.06 0.11 R-008380672-OOOE 2.02 0.02 R-008380534-OOOT 2.02 0.00 R-008380381-OOOJ 2.00 0.22 R-008380674-OOOX 1.97 0.02 R-008380725-OOOD 1.97 0.10 R-008380770-OOOX 1.97 0.11 R-008380369-OOOU 1.86 0.17 R-008380662-OOOM 1.86 0.07 R-008380333-OOOP 1.78 0.21 R-008380413-OOOP 1.73 0.23 R-008380748-OOOP 1.73 0.16 R-008380688-OOOZ 1.71 0.10 R-008380747-OOOF 1.70 0.18 R-008380337-OOOZ 1.62 0.15 R-008380711-OOOB 1.62 0.07 R-008380522-OOOH 1.61 0.17 R-008380308-OOOF 1.56 0.08 R-008380387-OOOL 1.54 0.03 R-008380720-OOOK 1.50 0.07 R-008380769-OOOH 1.48 0.21 R-008380404-OOOF 1.46 0.04 R-008380786-OOOS 1.46 0.03 R-008380630-OOOT 1.44 0.05 R-008380727-OOOW 1.41 0.19 R-008380779-OOOA 1.38 0.04 R-008380594-OOOX 1.34 0.05 R-008380438-OOOT 1.32 0.06 R-008380456-OOOK 1.32 0.00 R-008380768-OOOZ 1.30 0.00 R-008380600-OOOR 1.28 0.00 R-008380762-OOOX 1.25 0.18 R-008380370-OOOH 1.19 0.17 R-008380349-OOOJ 1.19 0.36 R-008380390-OOOT 1.19 0.31
R-008380447-000B 1.13 0.06 R-008380666-OOOX 1.12 0.25 R-008380639-OOOW 1.11 0.03 R-008380546-OOOC 1.05 0.07 R-008380685-OOOY 1.04 0.02 R-008380698-000S 1.04 0.09 R-008380414-OOOY 1.03 0.10 R-008380401-OOOE 1.02 0.24
[0354] A subset of siNAs from Table 6a having a large log2(fold change) in the primary screen were re-racked onto a single 96-well plate in triplicate and screened again in murine Heap1-6 cells transfected with pSiCheckHepB-partial in order to confirm knockdown and to directly compare siNAs that were synthesized on different plates. Table 6B. summarizes the data. The siNAs were ranked based on log2(fold change) in HBV mRNA expression levels.
Table 6B. Log2(fold change) of various siNAs tested on a re-racked plate
Mean log 2(fold SD log 2(fold siNA Duplex ID change) change) R-008380665-OOON 6.79 0.09 R-008380648-OOOE 6.53 0.09 R-008380633-OOOU 6.45 0.05 R-008380783-OOOR 6.45 0.06 R-008380408-OOOR 6.36 0.05 R-008380645-OOOD 6.34 0.04 R-008380767-OOOR 6.32 0.01 R-008380764-OOOP 6.26 0.18 R-008380406-OOOY 6.23 0.04 R-008380740-OOOV 6.21 0.14 R-008380730-OOOC 6.19 0.06 R-008380362-OOOH 6.17 0.06 R-008380340-OOOF 6.12 0.10 R-008380558-OOOM 6.11 0.09 R-008380389-OOOD 6.10 0.22
R-008380363-OOOS 6.09 0.06 R-008380777-OOOH 6.08 0.07 R-008380772-OOOP 6.02 0.03 R-008380576-OOOE 6.00 0.05 R-008380771-OOOF 5.99 0.03 R-008380359-OOOB 5.99 0.12 R-008380377-OOOU 5.97 0.06 R-008380763-OOOF 5.94 0.11 R-008380680-OOOE 5.90 0.01 R-008380776-OOOZ 5.86 0.04 R-008380793-OOOH 5.86 0.04 R-008380417-OOOZ 5.85 0.07 R-008380785-OOOH 5.83 0.04 R-008380750-OOOM 5.63 0.02 R-008380420-OOOF 5.10 0.09 R-008380782-OOOG 4.47 0.31 R-008380753-OOON 4.45 0.15 R-008380689-OOOH 4.42 0.04 R-008380755-OOOF 4.36 0.13 R-008380736-OOOE 4.35 0.05 R-008380346-OOOH 4.35 0.02 R-008380756-OOOP 4.32 0.01 R-008380734-OOOM 4.21 0.09 R-008380781-OOOY 4.17 0.03 R-008380737-OOON 4.03 0.26 R-008380757-OOOY 3.97 0.06 R-008380791-OOOR 3.89 0.03 R-008380528-OOOK 3.78 0.06 R-008380787-OOOA 3.65 0.13 R-008380686-OOOG 3.63 0.16 R-008380754-OOOX 3.54 0.08 R-008380597-OOOY 3.49 0.07 R-008380321-OOOE 3.47 0.09 R-008380567-OOOW 3.43 0.05 R-008380624-OOOK 3.27 0.08 R-008380789-OOOT 3.20 0.09 R-008380746-OOOX 3.18 0.03
R-008380778-000S 3.13 0.10 R-008380735-000W 3.08 0.09 R-008380399-OOOW 2.82 0.02 R-008380410-OOON 2.60 0.08 R-008380311-000M 1.24 0.07 R-008380353-OOOZ 1.11 0.03 R-008380348-OOOA 0.75 0.04 R-008380315-OOOX 0.69 0.06
[0355] Select high ranking (larger log 2(fold change) value) siNAs from Table 6b were further analyzed for efficacy and potency in Hepal-6 cells transfected with pSiCheck-HepB partial Plasmid. Cells were treated with a 1:6 serial dose titration of siNA in order to establish a value. The Potency50 is the calculated siNA transfection concentration that produces 50% target mRNA knockdown. The results for these siNAs are shown in Table 6c.
Table 6c: Dose response data for various siNAs in Hepal-6 murine hepatoma cell line transfected with pSiCheck-HepB-partial. Calculated maximum log 2(fold change) is determined from the dose response curve.
ssDRC max log Potency50 siNA Duplex ID 2(fold change) (nM) R-008380648-OOOE 7.78 0.104 R-008380783-OOOR 7.01 0.023 R-008380665-OOON 6.97 0.008 R-008380645-OOOD 6.92 0.032 R-008380408-OOOR 6.80 0.016 R-008380633-OOOU 6.79 0.012 R-008380767-OOOR 6.72 0.028 R-008380730-OOOC 6.58 0.038 R-008380777-OOOH 6.57 0.020 R-008380740-OOOV 6.55 0.019 R-008380558-OOOM 6.45 0.014
R-008380406-OOOY 6.32 0.006 R-008380764-OOOP 6.27 0.030 R-008380772-OOOP 6.26 0.024 R-008380389-OOOD 6.22 0.021 R-008380362-OOOH 6.17 0.014 R-008380363-000S 5.87 0.009 R-008380340-OOOF 5.76 0.016 R-008380689-OOOH 5.30 0.036 R-008380756-OOOP 5.00 0.186 R-008380736-OOOE 4.96 0.076 R-008380757-OOOY 4.88 0.216 R-008380753-OOON 4.87 0.184 R-008380737-OOON 4.69 0.142 R-008380734-OOOM 4.65 0.226 R-008380791-OOOR 4.37 0.101
Example 2: Determining In Vitro Serum Stability of siNAs
[0356] siNAs are reconstituted as 50 M to 100 M stock solution with H 2 0 and added to human serum pre-warmed to 37°C to a final concentration of 20[g/mL. The mixture is then incubated at 37°C for 0, 1 and 2 hours. At the end of each time point, the reactions are stopped by mixing with equal volume of Phenomenex Lysis-Loading Buffer. Oligonucleotides are purified in 96-well format by Phenomenex Solid Phase Extraction and lyophilized until dry with Labconco Triad Lyo-00417. The lyophilized samples are reconstituted in 150 pL of 1mM EDTA prepared with RNase-free H 20. The sample solutions re then diluted 5 fold with 1mM EDTA for liquid chromatography/mass spectrometry (LC/MS) analysis on ThermoFisher Orbitrap. Serum metabolites of the siNAs were determined based on the measured molecular weights.
Example 3: Testing of Cytokine Induction
[0357] To assess immunostimulative effects of various siNAs of the invention loaded in lipid nanoparticles (e.g., DLinDMA/Cholesterol/ S-PEG-C-DMA/DSPC in a 40/48/2/10 ratio),
C57B1/6 mice are dosed with a single 3mpk dose of LNP formulated siNAs through tail vein injection. Serum or plasma samples are collected at 3 and 24 hours post-dose. The cytokine and chemokine levels in these samples is measured with the SearchLight IR Cytokine Array from Aushon Biosciences according to the manufacturer's instruction. The cytokines and chemokines measured are IL-la, IL-1f, IL-6, KC, IL-10, IFNy, TNF, GMCSF, MIP-1, MCP-1/JE, and RANTES.
Example 4: Efficacy Studies in Mouse.
[0358] In vivo efficacy studies are conducted in SCID mice with humanized livers that are subsequently inject with HBV. Transgenic mice with diseased livers are injected with human primary hepatocytes that allows repopulation of the mouse liver with up to 70% human hepatocytes. Mice are then inoculated with human HBV isolates. Infected mice are dosed IV via tail vein injections with LNP encapsulated siNAs or vehicle control using a single 3mpk dose. Effect of siNA on viral load is determined taking serum sample at various time points post dosing and then measuring the level of circulating HBV genomes in the serum by qPCR.
Example 5: Pharmacodynamic Study in Non-Human Primates
[0359] HBV-infected chimpanzees are dosed with a single 2.5mpk dose of siNA loaded lipid nanoparticles through intravenous infusion. To monitor HBV virus load, serum samples are taken at various time points pre- and post-dose and the level of HBV genomes are determined via qPCR. All procedures adhere to the regulations outlined in the USDA Animal Welfare Act (9 CFR, Parts 1, 2 and 3) and the conditions specified in The Guide for Care and Use of Laboratory Animals (ILAR publication, 1996, National Academy Press).
Example 6: siNAs LNP Formulations
A. GeneralLNP ProcessDescriptionfor DLinDAL4 Formulations:
[0360] The lipid nanoparticles are prepared by an impinging jet process. The particles are formed by mixing lipids dissolved in alcohol with siNA dissolved in a citrate buffer. The mixing ratio of lipids to siNA is targeted at 45-55% lipid and 65-45% siNA. The lipid solution contains a cationic lipid, a helper lipid (cholesterol), PEG (e.g. PEG-C-DMA, PEG-DMG) lipid, and DSPC at a concentration of 5-15 mg/mL with a target of 9-12 mg/mL in an alcohol (for example ethanol). The ratio of the lipids has a mole percent range of 25-98 for the cationic lipid with a target of 35-65, the helper lipid has a mole percent range from 0-75 with a target of 30-50, the PEG lipid has a mole percent range from 1-15 with a target of 1-6, and the DSPC has a mole percent range of 0-15 with a target of 0-12. The siNA solution contains one or more siNA sequences at a concentration range from 0.3 to 1 .0 mg/mL with a target of 0.3 -0.9 mg/mL in a sodium citrate buffered salt solution with pH in the range of 3.5-5. The two liquids are heated to a temperature in the range of 15-40°C, targeting 30-40°C, and then mixed in an impinging jet mixer instantly forming the LNP. The teelD has a range from 0.25 to 1.0 mm and a total flow rate from 10-600 m/minute The combination of flow rate and tubing ID has the effect of controlling the particle size of the LNPs between 30 and 200 nm. The solution is then mixed with a buffered solution at a higher pH with a mixing ratio in the range of 1:1 to 1:3 vol:vol but targeting 1:2 vol:vol. This buffered solution is at a temperature in the range of 15-40°C, targeting 30-40°C. The mixed LNPs are held from 30 minutes to 2 hrs prior to an anion exchange filtration step. The temperature during incubating is in the range of 15-40°C, targeting -40°C. After incubating, the solution are filtered through a 0.8 um filter containing an anion exchange separation step. This process uses tubing IDs ranging from 1 mm ID to 5 mm ID and a flow rate from 10 to 2000 m/minute The LNPs are concentrated and diafiltered via an ultrafiltration process where the alcohol is removed and the citrate buffer is exchanged for the final buffer solution such as phosphate buffered saline. The ultrafiltration process uses a tangential flow filtration format (TFF). This process uses a membrane nominal molecular weight cutoff range from 30 -500 KD. The membrane format is hollow fiber or flat sheet cassette. The TFF processes with the proper molecular weight cutoff retains the LNP in the retentate and the filtrate or permeate contains the alcohol; citrate buffer; and final buffer wastes. The TFF process is a multiple step process with an initial concentration to a siNA concentration of 1 -3 mg/mL. Following concentration, the LNPs solution is diafiltered against the final buffer for 10 -20 volumes to remove the alcohol and perform buffer exchange. The material is then concentrated an additional 1-3 fold. The final steps of the LNP process are to sterilefilter the concentrated LNP solution and vial the product.
Analytical Procedure:
1) siNA concentration
[0361] The siNA duplex concentrations are determined by Strong Anion-Exchange High Performance Liquid Chromatography (SAX-HPLC) using Waters 2695 Alliance system (Water Corporation, Milford MA) with a 2996 PDA detector. The LNPs, otherwise referred to as RNAi Delivery Vehicles (RDVs), are treated with 0.5% Triton X-100 to free total siNA and analyzed by SAX separation using a Dionex BioLC DNAPac PA 200 (4 x 250 mm) column with UV detection at 254 nm. Mobile phase is composed of A: 25 mM NaClO 4 , 10mM Tris, 20% EtOH, pH 7.0 and B: 250 mM NaClO 4 , 10 mM Tris, 20% EtOH, pH 7.0 with a liner gradient from 0-15 min and a flow rate of 1 ml/minute. The siNA amount is determined by comparing to the siNA standard curve.
2) Encapsulation rate
[0362] Fluorescence reagent SYBR Gold is employed for RNA quantitation to monitor the encapsulation rate of RDVs. RDVs with or without Triton X-100 are used to determine the free siNA and total siNA amount. The assay is performed using a SpectraMax M5e microplate spectrophotometer from Molecular Devices (Sunnyvale, CA). Samples are excited at 485 nm and fluorescence emission is measured at 530 nm. The siNA amount is determined by comparing to an siNA standard curve.
Encapsulation rate = (1- free siNA/total siNA) x100%
3) Particle size and polydispersity
[0363] RDVs containing 1 g siNA are diluted to a final volume of 3 ml with 1 x PBS. The particle size and polydispersity of the samples is measured by a dynamic light scattering method using ZetaPALS instrument (Brookhaven Instruments Corporation, Holtsville, NY). The scattered intensity is measured with He-Ne laser at 25°C with a scattering angle of 90°.
4) Zeta Potential Analysis
[0364] RDVs containing 1 g siNA are diluted to a final volume of 2 ml with 1 mM Tris buffer (pH 7.4). Electrophoretic mobility of samples is determined using ZetaPALS instrument (Brookhaven Instruments Corporation, Holtsville, NY) with electrode and He-Ne laser as a light source. The Smoluchowski limit is assumed in the calculation of zeta potentials.
5) Lipid analysis
[0365] Individual lipid concentrations are determined by Reverse Phase High Performance Liquid Chromatography (RP-HPLC) using Waters 2695 Alliance system (Water Corporation, Milford MA) with a Corona charged aerosol detector (CAD) (ESA Biosciences, Inc, Chelmsford, MA). Individual lipids in RDVs are analyzed using an Agilent Zorbax SB-C18 (50 x 4.6 mm, 1.8 pm particle size) column with CAD at 60°C. The mobile phase is composed of A: 0.1% TFA in H 2 0 and B: 0.1% TFA in IPA. The gradient changes from 60% mobile phase A and 40% mobile phase B from time 0 to 40% mobile phase A and 60% mobile phase B at 1.00 min; 40% mobile phase A and 60% mobile phase B from 1.00 to 5.00 min; 40% mobile phase A and 60% mobile phase B from 5.00 min to 25% mobile phase A and 75% mobile phase B at 10.00 min; 25% mobile phase A and 75% mobile phase B from 10.00 min to 5% mobile phase A and 95% mobile phase B at 15.00 min; and 5% mobile phase A and 95% mobile phase B from 15.00 to 60% mobile phase A and 40% mobile phase B at 20.00 min with a flow rate of 1 ml/minute. The individual lipid concentration is determined by comparing to the standard curve with all the lipid components in the RDVs with a quadratic curve fit. The molar percentage of each lipid is calculated based on its molecular weight.
B. General Formulation Procedure for CLinDMA/Cholesterol/PEG-DMG at a ratio of 71.9:20.2:7.9.
[0366] siNA solutions were prepared by dissolving siNAs in 25mM citrate buffer (pH 4.0) at a concentration of 0.8mg/mL. Lipid solutions were prepared by dissolving a mixture of 2S Octyl-ClinDMA, cholesterol and PEG-DMG at a ratio of 71.9:20.2:7.9 in absolute ethanol at a concentration of about 10mg/mL. Equal volume of siNA and lipid solutions were delivered with two syringe pumps at the same flow rates to a mixing T connector. The resulting milky mixture was collected in a sterile bottle. This mixture was then diluted slowly with an equal volume of citrate buffer, and filtered through a size exclusion hollow fiber cartridge to remove any free siNA in the mixture. Ultra filtration against citrate buffer (pH 4.0) was employed to remove ethanol (test stick from ALCO screen), and against PBS (pH 7.4) to exchange buffer. The final LNP was obtained by concentrating to a desired volume and sterile filtered through a 0.2mm filter. The obtained LNPs were characterized in term of particle size, alcohol content, total lipid content, nucleic acid encapsulated, and total nucleic acid concentration.
C. General LNP PreparationFor Various Formulations in Table 10
[0367] siNA nanoparticle suspensions in Table 10 were prepared by dissolving siNAs and/or carrier molecules in 20 mM sodium citrate buffer (pH 5.0) at a concentration of about 0.40 mg/mL. Lipid solutions were prepared by dissolving a mixture of cationic lipid (e.g., (13Z,16Z) N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine, see structure in Table 11), DSPC, Cholesterol, and PEG-DMG (ratios shown in Table 10) in absolute ethanol at a concentration of about 8 mg/mL. The nitrogen to phosphate ratio was approximated to 6:1.
[0368] Nearly equal volumes of siNA/carrier and lipid solutions were delivered with two FPLC pumps at the same flow rates to a mixing T connector. A back pressure valve wais used to adjust to the desired particle size. The resulting milky mixture was collected in a sterile glass bottle. This mixture was then diluted with an equal volume of citrate buffer, followed by equal volume of PBS (pH 7.4), and filtered through an ion-exchange membrane to remove any free siNA/carrier in the mixture. Ultra filtration against PBS (7.4)) was employed to remove ethanol and to exchange buffer. The final LNP was obtained by concentrating to the desired volume and sterile filtered through a 0.2 m filter. The obtained LNPs were characterized in term of particle size, Zeta potential, alcohol content, total lipid content, nucleic acid encapsulated, and total nucleic acid concentration.
LNP ManufactureProcess
[0369] In a non-limiting example, LNPs were prepared in bulk as follows. The process consisted of (1) preparing a lipid solution; (2) preparing an siNA/carrier solution; (3) mixing/particle formation; (4) incubation; (5) dilution; (6) ultrafiltration and concentration.
1. Preparation of Lipid Solution
[0370] 2L glass reagent bottles and measuring cylinders were depyrogenated. The lipids were warmed to room temperature. Into the glass reagent bottle was transferred 8.Og of (13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine with a pipette and 1.2g of DSPC, 3.5g of Cholesterol, 0.9g of PEG-DMG were added. To the mixture is added IL of ethanol. The reagent bottle was placed in heated water bath, at a temperature not exceeding 50°C. The lipid suspension was stirred with a stir bar. A thermocouple probe was put into the suspension through one neck of the round bottom flask with a sealed adapter. The suspension was heated at -40 °C until it became clear. The solution was allowed to cool to room temperature.
2. Preparation of siNA/Carrier Solution
[0371] Into a sterile container (Coming storage bottle) was weighed 0.4 g times the water correction factor (approximately 1.2) of siNA powder. The siNA was transferred to a depyrogenated 2 L glass reagent bottle. The weighing container was rinsed 3x with citrate buffer (20mM, pH 5.0) and the rinses were placed into the 2 L glass bottle, QS with citrate buffer to 1 L. The concentration of the siNA solution was determined with a UV spectrometer using the following procedure. 20 pL was removed from the solution, diluted 50 times to 1000 [L, and the UV reading recorded at A260 nm after blanking with citrate buffer. This was repeated. Note, if the readings for the two samples are consistent, an average can be taken and the concentration calculated based on the extinction coefficients of the siNAs. If the final concentration is out of the range of 0.40 0.01 mg/mL, the concentration can be adjusted by adding more siNA/carrier powder, or adding more citrate buffer. This process can be repeated for the second siNA, if applicable
[0372] When the siNA/carrier solution comprised a single siNA duplex instead of a cocktail of two or more siNA duplexes and/or carriers, then the siNA/carrier was dissolved in 20 mM citrate buffer (pH 5.0) to give afinal concentration of 0.4 mg/mL.
[0373] The lipid and ethanol solutions were then sterile filtered through a Pall Acropak 20 0.8/0.2 pm sterile filter PN 12203 into a depyrogenated glass vessel using a Master Flex Peristaltic Pump Model 7520-40 to provide a sterile starting material for the encapsulation process. The filtration process was run at an 80 mL scale with a membrane area of 20 cm2 . The flow rate was 280 mL/minute. This process can be scaled by increasing the tubing diameter and the filtration area.
3. Particle formation - Mixing step
[0374] Using a two-barrel syringe driven pump (Harvard 33 Twin Syringe), the sterile lipid/ethanol solution and the sterile siNA/carrier or siNA/carrier cocktail/citrate buffer (20 mM citrate buffer, pH 5.0) solutions were mixed in a 0.5mm ID T-mixer (Mixing Stage I) at equal, or nearly equal, flow rates. The resulting outlet LNP suspension contained 40-50 vol% ethanol. To obtain a 45 vol% ethanol outlet suspension, the sterile lipid/ethanol and the sterile siNA/carrier or siNA/carrier cocktail/citrate buffer solutions were mixed at flow rates of 54 mL/min and 66 mL/min, respectively, such that the total flow rate of the mixing outlet is 120 m/min.
4. Dilution
[0375] The outlet stream of Mixing Stage I was fed directly into a 4mm ID T-mixer (Mixing Stage II), where it was diluted with a buffered solution at higher pH (20 mM sodium citrate, 300 mM sodium chloride, pH 6.0) at a ratio of 1:1 vol:vol %. This buffered solution was at a temperature in the range of 30-40°C, and was delivered to the 4mm T-mixer via a peristaltic pump (Cole Parmer MasterFlex L/S 600 RPM) at a flow rate of 120 mL/min.
[0376] The outlet stream of Mixing Stage II was fed directly into a 6mm ID T-mixer (Mixing Stage III), where it was diluted with a buffered solution at higher pH (PBS, pH 7.4) at a ratio of 1:1 vol:vol%. This buffered solution was at a temperature in the range of 15-25°C, and was delivered to the 6mm T-mixer via peristaltic pump (Cole Parmer MasterFlex L/S 600 RPM) at a flow rate of 240 mL/min.
5. Incubation and Free siNA Removal
[0377] The outlet stream of Mixing Stage III was held after mixing for 30 minute incubation. The incubation was conducted at temperature of 35-40°C and the in-process suspension was protected from light. Following incubation, free (un-encapsulated) siNA was removed via anion exchange with Mustang Q chromatography filters (capsules). Prior to use, the chromatography filters were pre-treated sequentially with flushes of IN NaOH, IM NaCl, and a final solution of 12.5 vol% ethanol in PBS. The pH of the final flush was checked to ensure pH <8. The incubated LNP stream was then filtered via Mustang Q filters via peristaltic pump (Cole Parmer MasterFlex L/S 600 RPM) at flow rate of approximately 100 mL/min. The filtered stream was received into a sterile glass container for ultrafiltration and concentration as follows.
6. Ultrafiltration, Concentration and Sterile Filtration
[0378] The ultrafiltration process is a timed process and the flow rates must be monitored carefully. This is a two step process; thefirst is a concentration step taking the diluted material and concentrating approximately 8-fold, to a concentration of approximately 0.3-0.6 mg/mL siNA.
[0379] In the first step, a ring-stand with a ultrafiltration membrane 100 kDa PES (Spectrum Labs) installed was attached to a peristaltic pump (Spectrum KrosFloII System). 9.2 L of sterile distilled water was added to the reservoir; 3 L was drained to waste and the remainder was drained through permeate to waste. 5.3 L of 0.25 N sodium hydroxide was added to the reservoir with 1.5 L drained to waste and 3.1 L drained through permeate to waste. The remaining sodium hydroxide was held in the system for sanitization (at least 10 minutes), and then the pump was drained. 9.2 L of 70 (v/v)% isopropyl alcohol was added to the reservoir with 1.5 L drained to waste and the remainder drained through permeate to waste. 6 L of conditioning buffer (12.5% ethanol in phosphate buffered saline) was added with 1.5 L drained to waste and the remainder drained though the permeate until the waste was of neutral pH (7-8). A membrane flux value was recorded, and the pump was then drained.
[0380] The diluted LNP solution was placed into the reservoir to the 1.1 L mark. The pump was turned on at 2.3 L/min. After 5 minutes of recirculation, the permeate pump was turned on at 62.5 mL/min and the liquid level was constant at approximately 950 mL in the reservoir. The diluted LNP solution was concentrated from 9.8 L to 1.1 L in 140 minutes, and the pump was paused when all the diluted LNP solution has been transferred to the reservoir.
[0381] The second step was a diafiltration step exchanging the ethanol/aqueous buffer to phosphate buffered saline. During this step, approximately 10-20 diafiltration volumes of phosphate buffered saline were used. Following diafiltration, a second concentration was undertaken to concentrate the LNP suspension 3-fold to approximately 1-1.5 mg/mL siRNA. The concentrated suspension was collected into sterile, plastic PETG bottles. The final suspension was then filtered sequentially via Pall 0.45 um PES and Pall 0.2 um PES filters for terminal sterilization prior to vial filling.
[0382] The obtained LNPs were characterized in terms of particle size, Zeta potential, alcohol content, total lipid content, nucleic acid encapsulated, and total nucleic acid concentration.
D. Synthesis of novel cationic lipids
[0383] Synthesis of the novel cationic lipids is a linear process starting from lipid acid (i). Coupling to N,O-dimethyl hydroxylamine gives the Weinreb amide ii. Grignard addition generates ketone iii. Titanium mediated reductive amination gives final products of type iv.
GENERAL SCHEME 1
0 00
HO L, CH 3 NH(OCH 3 ) N L, L2 MgBr, THF LkL, EDC, HOBT,TEA,DCM | ii iii
R 1R 2NH, Ti(OiPr) 4 , NaBH 4 N R2
L2 L,
iv
[0384] Synthesis of the single carbon homologated cationic lipids v is a linear process starting from lipid ketone (iii). Conversion of the ketone to the nitrile (iv) is accomplished via treatment with TOSMIC and potassium tert-butoxide. Reduction of the nitrile to the primary amine followed by reductive amination provides final cationic lipids v.
GENERAL SCHEME 2
0 CN TOSMIC 1. LAH R L2 L1 K'tBuO-, DME L2 L1 2. RCHO, Na(AcO) 3BH Li Lj iii iv V
[0385] Synthesis of two carbon homologated cationic lipids viii is a linear process starting from lipid ketone (iii). Conversion of the ketone to the af,p-unsaturated amide vi is accomplished under Peterson conditions. Conjugate reduction of the aP-unsaturation is performed using LS-Selectride to give amide vii. Reduction of the amide with lithium aluminum hydride provides final cationic lipids viii.
GENERAL SCHEME 3
00 0
0N N. N O N IN LS-Selectride N LAH N L2 L, nBuLi, THF L L, L2 L, L2 L,
iii vi vii viii
[0386] Cyclopropyl containing lipids are prepared according to General Scheme 4. Unsaturated Weinreb amides ii are subjected to Simmons-Smith cyclopropanation conditions to give cyclopropyl containing Weinreb amides ix. These are carried on to final products as outlined in General Schemes 1-3.
GENERAL SCHEME 4 0 0
ON (Et) 2Zn, CH 2 12 ON n n TFA, DCM n n ii ix
[0387] Synthesis of allylic amine cationic lipids xv is a linear process starting with aldehyde x. Addition of t-butyl aceate generates p-hydroxy ester xi. Conversion of the hydroxyl functionality to a fluoro group followed by acid treatment generates p-fluoro acid xii. Conversion of the acid to the Weinreb amide followed by Grignard addition gives the P-fluoro ketone xiv. Reductive amination results in simultaneous elimination to generate the desired allylic amine xv.
GENERAL SCHEME 5
O 10 OH 0 F t-butyl acetate 1. Deoxofluor CH 3 NH(OCH 3
) H R LDA, THF 0 R 2. HCI HO0' R EDC, HOBT, TEA, DCM x xi xii
O F 0 F RRNHTiCVD\ NIDJ R N R2
N R L2 MgBr, THF L2 R 12' iir 4 ,a 4 R / Xiii XIV XV
,23-nonacosadien-10-amine, N,N-dimethyl-, (20Z,23Z) (Compound 1)
0
CH 3 NH(OCH 3
ii
[0388] 11,14-Eicosadienoic acid, (11Z,14Z)- (50 g, 162 mmol), N,0 Dimethylhydroxylamine hydrochloride (31.6 g, 324 mmol), HOAt (44.1 g, 324 mmol), Et 3 N (45.2 mL, 324 mmol), and EDC (62.1 g, 324 mmol) were mixed in DCM (810 mL) and stirred overnight at ambient temperature. Reaction was then washed 5 x 700 mL water, then washed 1 x
600 mL 1 M NaOH, dried with sodium sulfate, filtered through celite and evaporated to obtain 53.06 g (93%) 11,14-eicosadienamide, N-methoxy-N-methyl-, (11Z,14Z) as a clear golden oil. H NMR (400 MHz, CDC 3) 65.35 (m, 4H), 3.68 (s, 3H), 3.18 (s, 3H), 2.77 (m, 2H), 2.41 (t, J=
7 Hz, 2H), 2.05 (m, 4H), 1.63 (m, 2H), 1.40-1.26 (m, 18H), 0.89 (t, J= 7 Hz, 3H).
O0N 1. C 9H1 9 MgBr, THF 2. (CH 3) 2 NH, Ti(OiPr)4 , NaBH 4 ii
[0389] 11,14-eicosadienamide, N-methoxy-N-methyl-, (11Z,14Z)- 1 (4 g, 11.38 mmol) was dissolved in dry THF (50.0 ml) in a 250 mL flask then1 M nonylmagnesium bromide (22.76 ml, 22.76 mmol) was added under nitrogen at ambient temperature. After 10 min, the reaction was slowly quenched with excess sat. aq NH 4Cl. The reaction was washed into a separatory funnel with hexane and water, shaken, the lower aqueous layer discarded, the upper layer dried with sodium sulfate, filtered, and evaporated to give crude ketone as a golden oil. To the above crude ketone was added dimethylamine (2 M in THF) (14.22 ml, 28.4 mmol) followed by Ti(O-i-Pr) 4 (6.67 ml, 22.76 mmol) and let stir overnight. The next day, added EtOH (50 ml) followed by NaBH 4 (0.646 g, 17.07 mmol). After 5 min of stirring, directly injected entire reaction onto a 40 g silica column that was in line with a 330 g silica column. Eluted 10 min 100% DCM, then 30 min 0-15% MeOH/DCM, collected 20,23-nonacosadien-10-amine, N,N-dimethyl-, (20Z,23Z) (1) (2.45 g, 5.47 mmol, 48.1 % yield) as a faintly golden oil. 'H NMR (400 MHz, CDCl 3) 6 5.35 (m, 4H), 2.78 (m, 2H), 2.23 (m, 1H), 2.21 (s, 6H), 2.05 (m, 4H), 1.45-1.16 (m, 38H), 0.89 (m, 6H). HRMS calcd for C31H61N 448.4877, found 448.4872.
[0390] Compounds 2-30 are novel cationic lipids and were prepared according to the General Scheme 1 above.
Compound Structure HRMS N
2 calcd C28H56N 406.4407, found 406.4405.
3 calcd C27H54N 392.4251, found 392.4250.
4 calcd C24H48N 350.3781, found 350.3770.
calcd C23H46N 336.3625, found 336.3613.
6 calcd C25H50N 364.3938, found 364.3941.
7 calcd C26H52N 378.4094, found 378.4081.
8 calcd C29H58N 420.4564, found 420.4562.
9 calcd C26H52N 378.4094, found 378.4089.
calcd C25H50N 364.3938, found 364.3931.
11 calcd C30H60N 434.4720, found 434.4717.
12 calcd C29H58N 420.4564, found 420.4561.
13 calcd C28H56N 406.4407, found 406.4404.
14 calcd C27H54N 392.4251, found 392.4245.
calcd C33H66N 476.5190, found 476.5196.
16 calcd C32H64N 462.5033, found 462.5045.
17 calcd C29H59N 422.4720, found 422.4726.
18 calcd C28H57N 408.4564, found408.4570.
19 calcd C30H59N 434.4720, found 434.4729.
calcd C29H61N 424.4877, found 424.4875.
21 calcd C32H64N 462.5033, found 462.5023.
22 calcd C33H64N 474.5033, found 474.5033.
23 calcd C29H60N 422.4720, found 422.4716.
24 calcd C29H60N 422.4720, found 422.4718.
25 calcd C31H64N 450.5033, found 450.5031.
26 calcd C31H64N 450.5033, found 450.5034.
27 calcd C35H72N 506.5659, found 506.5635.
28 calcd C31H64N 450.5033, found 450.5037.
29 calcd C33H68N 478.5346, found 478.5358.
30 calcd C27H56N 394.4407, found 394.4407.
(12Z,15Z)-N,N-dimethyl-2-nonylhenicosa-12,15-dien-1-amine (Compound 31)
0 TOSMIC K tBuO, DME
iv
[0391] A solution of keton iii (4.0g, 9.55mmol), TOSMIC (2.4g, 12.4mmol) in dimethoxyethane (45 mL) was cooled to 0 C and treated with potassium tert-butoxide (19.1mmol, 19.1mL of a IM solution in tBuOH). After 90 minutes, the reaction was partitioned between hexanes and water. The organics were washed with water, dried over sodium sulfate, filtered and evaporated in vacuo. This material was purified by flash chromatography (0-5%
EtOAc/hexanes) to give desired product (containing ~20% of s.m.). This mixture was carried into next step as is. LC/MS (M+H) = 430.6.
iv
H2N
[0392] Lithium aluminum hydride (23.9mmol, 23.9mL of a IM solution in THF) was added directly to nitrile iv (3.42g, 8mmol) at ambient temperature and the reaction was stirred for 20 minutes. The reaction was diluted with 1OOmL THF, cooled to 0 C and carefully quenched with sodium sulfate decahydrate solution. The solids were filtered off and washed with THF. The filtrate was evaporated in vacuo and carried directly into next reaction crude. LC/MS (M+H)= 434.6.
H2N HCHO, Na(AcO) 3BH
31
[03931 A solution of primary amine (3.45g, 6.2mmol) in dichloroethane (OOmL) was treated with formaldehyde (1.6mL, 21.7mmol) followed by sodium triacetoxyborohydride (6.6g, 31mmol). After 5 minutes, the reaction was partitioned between dichloromethane and IN
NaOH. The organics were dried over sodium sulfate, filtered and evaporated in vacuo. The crude mixture was purified by reverse phase preparative chromatography (C8 column) to provide (12Z,15Z)-N,N-dimethyl-2-nonylhenicosa-12,15-dien-1-amine. HRMS calc'd 462.5033, found 462.5026. 'H NMR (400 MHz, CDC 3) 6 5.35 (m, 4H), 2.78 (2H, t, J=5.6Hz), 2.18 (s, 6H), 2.05 (m, 6H), 1.3 (m, 39H), 0.89 (m, 6H).
(13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine (Compound 32)
0
iiii nBui TH
[0394] The silyl amide Peterson reagent (3.lg, 16.7mmol) was dissolved in THF (35mL) and cooled to -63 C. To this solution was added nBuLi (16.7mmol, 6.7mL of a 2.5M solution). The reaction was warmed to ambient temperature for 30 minutes. The ketone (5.0g, 11.9mmol) was dissolved in THF (25mL) in a second flask. The Peterson reagent was transferred to the ketone solution at -60 C. The reaction was warmed to -40 C for 1 hour, then warmed to 0 C for 30 minutes. The reaction was quenched with sodium bicarbonate, diluted with additional water and partitioned between water/hexanes. The organics were washed with brine, dried over sodium sulfate, filtered and evaporated in vacuo. Purification by flash chromatography (0-40% MTBE/hexanes) gave c,p-unsatured amide vi. H NMR (400 MHz, CDC 3) 6 5.75 (s, 1H), 5.36 (m, 4H), 3.01 (s, 3H), 2.99 (s, 3H), 2.78 (t, 2H), 2.28 (t, 2H), 2.05 (m, 6H), 1.35 (m, 34H), 0.89 (m, 6H).
N LS-Selectride
Vi
Vii
[0395] cp-unsatured amide vi(g, 2.1mmol) and LS-Selectride (4.1mmol, 4.lmL of a IM solution) were combined in a sealed tube and heated to 60 C for 24 hours. The reaction was cooled to ambient temperature and partitioned between ammonium chloride solution and heptane. The organics were dried over sodium sulfate, filtered and evaporated in vacuo to give amide vii. This intermediate was carried directly into next reaction crude.
Vii
32
[0396] To a solution of amide vii (2.85g, 5.8mmol) was added lithium aluminum hydride (8.7mmol, 8.7mL of a IM solution). The reaction was stirred at ambient temperature for 10 minutes then quenched by slow addition of sodium sulfate decahydrate solution. The solids were filtered and washed with THF and the filtrate evaporated in vacuo. The crude mixture was
purified by reverse phase preparative chromatography (C8 column) to provide (13Z,16Z)-NN dimethyl-3-nonyldocosa-13,16-dien-1-amine (Compound 32) as an oil. HRMS (M+H) calc'd
476.5190, found 476.5189. 'H NMR (400 MHz, CDCl3) 6 5.37 (m, 4H), 2.78 (t, 2H), 2.42 (m, 8H), 2.05 (q, 4H), 1.28 (m, 41H), 0.89 (m, 6H).
N,N-dimethyl-1-(2-octylcyclopropyl)heptadecan-8-amine (Compound 33)
0 CH 3NH(OCH 3
0 NN
ii
[0397] To a solution of oleic acid (1g, 3.5mmol) in DCM (500 mL) cooled to 0 C was added CDI (0.63g, 3.9mmol). The reaction was warmed to ambient temperature for 30 minutes before cooling to 0 C and treating first with triethylamine (0.39g, 3.9mmol) and then dimethyl hydroxylamine hydrochloride (0.38g, 3.9mmol). After 1 hour the reaction was partitioned between water and heptane. The organics were dried over magnesium sulfate, filtered and evaporate in vacuo to give crude Weinreb amide ii which was carried directly into next reaction.
0
N (Et) 2Zn, CH 2 12 TFA,DCM ii
0
ix
[0398] A solution of diethylzinc (70.3mmol, 70.3mL of a IM solution) in dichloromethane (130mL) was cooled to -1 C and treated dropwise with TFA (8.0g, 70.3mmol). After 30 minutes, diiodomethane (18.8g, 70.3mmol) was added and this was aged for 30 minutes in the ice bath. To this solution was added Weinreb amide ii (7.6g, 23.4mmol). The reaction was warmed to ambient temperature and stirred for 1 hour. The reaction was quenched with ammonium chloride solution (100mL) and organic layer partitioned off, washed with 10% sodium thiosulfate, dried over magnesium sulfate, filtered and evaporated in vacuo. Purification was flash chromatography (0-30% MTBE/heptane) gave desired product ix. 1H NMR (400 MHz, CDC 3) 6 3.72 (s, 3H), 3.22 (s, 3H), 2.48 (t, 2H), 1.65 (in, 2H), 1.39 (in,22H), 1.18 (in, 2H), 0.91 (t, 3H), 0.68 (in, 2H), 0.59 (in,1H), -0.32 (in,1H).
0
33
[0399] Conversion of Weinreb amide ix to Compound 33 was carried out in a manner analogous to that described for Compound 1 above (nonyl Grignard addition followed by reductive amination). LC/MS (M+H) = 436.6. 1H NMR (400 MHz, CDCl 3) 6 2.25 (s, 6H), 1.30 (in, 45H), 0.91 (in, 6H), 0.68 (in, 2H), 0.59 (in,1H), -0.31 (in,1H).
[0400] Compounds 34-43 are novel cationic lipids and were prepared according to General Schemes 1-4 above.
Compound Structure HRMS N
34 calcd C30H62N 436.4877, found 436.4872.
35 calcd C32H66N 464.5190, found 464.5186.
36 calcd C34H70N 492.5503, found 492.5496.
37 calcd C33H66N 476.5190, found 476.5174.
38 calcd C29H60N 422.4720, found 422.4701.
39 calcd C30H62N 436.4877, found 436.4880.
40 calcd C32H66N 464.5190, found 464.5199.
41 calcd C30H62N 436.4877, found 436.4877.
42 calcd C30H62N 436.4877, found 436.4875.
43 LC/MS (M+H) 408.6.
(11E,20Z,23Z)-NN-dimethylnonacosa-11,20,23-trien-10-amine (Compound 44)
0 t-butyl acetate LDA, THF x
xi
[0401] To a solution of LDA (95mmol, 47.5mL of a 2M solution) in THF (127mL) cooled to -78 C was added t-butyl acetate. The reaction was stirred for 15 minutes followed by addition of aldehyde x. The reaction was immediately quenched with ammonium chloride solution, warmed to ambient temperature and partitioned between water/pentane. The organics were dried over sodium sulfate, filtered and evaporated in vacuo. LC/MS (M+H-tBu)= 325.4.
O OH 1. Deoxofluor 2. HCI xi
xii
[0402] Hydroxy ketone xi (7g, 18.4mmol) was dissolved in dichloromethane (150mL) and cooled to 0 C and treated with deoxofluor (7.3g, 33.1mmol). The reaction was warmed to ambient temperature with stirring for 16 hours followed by quenching with sodium bicarbonate solution. The reaction was partitioned and the organics dried over sodium sulfate, filtered and evaporate in vacuo. Flash column chromotagraphy (0-5% ethyl acetate/hexanes) gave the fluoro ester.
[0403] Fluoro ester intermediate (6g, 15.6mmol) in dichloromethane was treated with hydrogen chloride (157mmol, 39.2mL of a 4M solution in dioxane) and the reaction was stirred at ambient temperature for 16 hours. The reaction was evaporated in vacuo to give desiredp fluoro acid xii. LC/MS (M+H) = 327.3.
O F CH 3 NH(OCH 3 )
EDC, HOBT, TEA, DCM xii
N xiii
[0404] Fluoro carboxylic acid xii (5.1g, 15.7mmol), EDC (6.0g, 31.4mmol), N,O dimethylhydroxylamine hydrochloride (3.1g, 31.4mmol), trimethylamine (4.0g, 39.2mmol), and HOAt (4.3g, 31.4mmol) were combined in DCM (78mL) and stirred at ambient temperature for 16 hours. The reaction was partitioned between water/DCM and the organics were washed with water (3x) and NaOH solution (lx), dried over sodium sulfate, filtered and evaporated in vacuo.
Crude material was purified by reverse phase preparative chromatography to give desired Weinreb amide xiii. LC/MS (M+H)= 370.4.
O F C9 HggMgBr, THF N xiii
[0405] A solution of Weinreb amide xiii (4.3g, 11.7mmol) in THF (50mL) was treated with nonylmagnesium bromide (23.4mmol, 23.4mL of a IM solution) at ambient temperature. The reaction was quenched with ammonium chloride solution after 1 hour and partitioned between water and pentane. The organics were dried over sodium sulfate, filtered and evaporated in vacuo. This material was carried into next step crude.
O F (CH 3 )2NH, Ti(OiPr)4 , NaBH 4
xiv
[0406] Ketone xiv (5.lg, 11.7mmol) was treated with dimethylamine (29.3mmol, 14.7mL of a 2M solution in THF) and titanium(IV) isopropoxide (6.7g, 23.5mmol) and the reaction was stirred at ambient temperature for 16 hours. To the reaction mixture was added ethanol (50mL) followed by sodium borohydride (0.67g, 17.6mmol). The reaction was loaded directly onto a silica column and purified by flash chromatography (0-15% MeOH/DCM). The material required a second purification by preparative reverse phase chromatography to give (11E,20Z,23Z)-NN-dimethylnonacosa-11,20,23-trien-10-amine. HRMS calc'd446.4720, found
446.4724. 'H NMR (400 MHz, CDC 3) 65.48 (in, H), 5.37 (in, 4H), 5.23 (in, H), 2.78 (t, 2H), 2.58 (in, 1H), 2.22 (s, 6H), 2.04 (in, 6H), 1.56 (in, H), 1.30 (in, 31H), 0.89 (in, 6H).
[0407] Compound 45 is DLinKC2DMA as described in Nature Biotechnology, 2010, 28, 172-176, WO 2010/042877 Al, WO 2010/048536 A2, WO 2010/088537 A2, and WO 2009/127060 Al.
0 (45)
[0408] Compound 46 is MC3 as described in WO 2010/054401, and WO 2010/144740 Al.
(46)
E. LipidNanoparticleCompositions
[0409] The following lipid nanoparticle compositions (LNPs) of the instant invention are useful for the delivery of oligonucleotides, specifically siNA molecules of the invention:
Cationic Lipid / Cholesterol / PEG-DMG 56.6/38/5.4; Cationic Lipid / Cholesterol / PEG-DMG 60/38/2; Cationic Lipid/ Cholesterol /PEG-DMG 67.3/29/3.7; Cationic Lipid / Cholesterol /PEG-DMG 49.3/47/3.7; Cationic Lipid / Cholesterol /PEG-DMG 50.3/44.3/5.4; Cationic Lipid / Cholesterol /PEG-C-DMA / DSPC 40/48/2/10; Cationic Lipid / Cholesterol /PEG-DMG / DSPC 40/48/2/10; and Cationic Lipid / Cholesterol /PEG-DMG / DSPC 58/30/2/10.
[0410] One skilled in the art would readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those inherent therein. The methods and compositions described herein, as presently representative of preferred embodiments, are exemplary and are not intended as limitations on the scope of the invention. Changes therein and other uses will occur to those skilled in the art, which are encompassed within the spirit of the invention, are defined by the scope of the claims.
Table 7: HBV Accession Numbers
X02763 - SEQ ID NO:504 Hepatitis b virus genome (serotype adw2) X02763.1 GI:59418
X69798 - SEQ ID NO:505 Hepatitis B virus, subtype adw4 genes X69798.1 GI:59422
X65259 - SEQ ID NO:506 Hepatitis B virus (ayw,patient E) genes PreS1, PreS2, PreC, C, X and polymerase X65259.1 GI:59439
X04615 - SEQ ID NO:507 Hepatitis B virus genome, subtype ayr X04615.1 GI:59585
D00329 - SEQ ID NO:508 Hepatitis B virus subtype ADW genomic DNA, complete genome, clone: pJDW233 D00329.1 GI:221497
M32138 - SEQ ID NO:509 Hepatitis B virus variant HBV-alphal, complete genome M32138.1 GI:329667
X75657 - SEQ ID NO:510 Human hepatitis virus (genotype E, Bas) preS1, preS2, S, C, X, antigens, core antigen, X protein and polymerase X75657.1 GI:452617
M12906 - SEQ ID NO:511 Hepatitis B virus subtype adr, complete genome. M12906.1 GI:474959
X85254 - SEQ ID NO:512 Hepatitis B virus genome (PreS1, PreS2, S, PreC, C, X genes and polymerase) X85254.1 GI:736003
X51970 - SEQ ID NO:513 Hepatitis B virus (HBV 991) complete genome. X51970.1 GI:1155012
AB014381 - SEQ ID NO:514 Hepatitis B virus genomic DNA, complete sequence, isolate 22Y4HCC AB014381.1 GI:3582357
AF100309 - SEQ ID NO:515 Hepatitis B virus strain 56, complete genome AF100309.1 GI:4323201
AF090842 - SEQ ID NO:516 Hepatitis B virus strain G5.27295, complete genome AF090842.1 GI:5114084
AB033554 - SEQ ID NO:517 Hepatitis B virus DNA, complete genome, isolate:RTB299 AB033554.1 GI:6063442
AB032431 - SEQ ID NO:518 Hepatitis B virus genomic DNA, complete sequence, isolate:HBV/E-Ch195 AB032431.1 GI:6691492
AF160501 - SEQ ID NO:519 Hepatitis B virus strain IG29227, complete genome AF160501.1 GI:6983934
AB036910 - SEQ ID NO:520 Hepatitis B virus (genotype F) genomic DNA, complete genome, isolate:VNZ8251 AB036910.1 GI:11191875
AF223965 - SEQ ID NO:521 Hepatitis B virus strain C-1858 isolate sa16, complete genome AF223965.1 GI:12247041
AB064310 - SEQ ID NO:522 Hepatitis B virus DNA, complete genome, clone:USG769 AB064310.1 GI:18146661
AF405706 - SEQ ID NO:523 Hepatitis B virus isolate 235/01, complete genome AF405706.1 GI:19849032
AY090454 - SEQ ID NO:524 Hepatitis B virus strain 1853Nic, complete genome AY090454.1 GI:22135696
AY090457 - SEQ ID NO:525 Hepatitis B virus strain 2928Nic, complete genome AY090457.1 GI:22135711
AY090460 - SEQ ID NO:526 Hepatitis B virus strain LAS2523, complete genome
AY090460.1 GI:22135726
Table 8
Non-limiting examples of Stabilization Chemistries for chemically modified siNA constructs
Chemistry pyrimidine purine caps p=S Strand
"Stab 00" Ribo Ribo TT at 3'- S/AS ends "Stab 1" Ribo Ribo - 5 at 5'-end S/AS 1 at 3'-end "Stab 2" Ribo Ribo - All linkages Usually AS "Stab 3" 2'-fluoro Ribo - 4 at 5'-end Usually S 4 at 3'-end "Stab 4" 2'-fluoro Ribo 5' and3'- - Usually S ends "Stab 5" 2'-fluoro Ribo - 1at 3'-end Usually AS "Stab 6" 2'-O-Methyl Ribo 5' and3'- - Usually S ends "Stab 7" 2'-fluoro 2'-deoxy 5' and3'- - Usually S ends "Stab 8" 2'-fluoro 2'-0- - 1 at 3'-end S/AS Methyl "Stab 9" Ribo Ribo 5' and3'- - Usually S ends "Stab 10" Ribo Ribo - 1at 3'-end Usually AS "Stab 11" 2'-fluoro 2'-deoxy - 1at 3'-end Usually AS "Stab 12" 2'-fluoro LNA 5' and 3'- Usually S ends "Stab 13" 2'-fluoro LNA 1 at 3'-end Usually AS "Stab 14" 2'-fluoro 2'-deoxy 2 at 5'-end Usually AS 1 at 3'-end "Stab 15" 2'-deoxy 2'-deoxy 2 at 5'-end Usually AS 1 at 3'-end "Stab 16" Ribo 2'-0- 5' and 3'- Usually S Methyl ends "Stab 17" 2'-O-Methyl 2'-0- 5' and 3'- Usually S Methyl ends "Stab 18" 2'-fluoro 2'-0- 5' and 3'- Usually S Methyl ends "Stab 19" 2'-fluoro 2'-0- 3'-end S/AS Methyl "Stab 20" 2'-fluoro 2'-deoxy 3'-end Usually AS "Stab 21" 2'-fluoro Ribo 3'-end Usually AS "Stab 22" Ribo Ribo 3'-end Usually AS "Stab 23" 2'-fluoro* 2'-deoxy* 5' and 3'- Usually S ends "Stab 24" 2'-fluoro* 2'-0-- 1 at 3'-end S/AS Methyl* "Stab 25" 2'-fluoro* 2'-0-- 1 at 3'-end S/AS Methyl* "Stab 26" 2'-fluoro* 2'-0-- S/AS Methyl* "Stab 27" 2'-fluoro* 2'-0- 3'-end S/AS Methyl* "Stab 28" 2'-fluoro* 2'-0- 3'-end S/AS Methyl* "Stab 29" 2'-fluoro* 2'-0- 1 at 3'-end S/AS Methyl* "Stab 30" 2'-fluoro* 2'-0- S/AS Methyl* "Stab 31" 2'-fluoro* 2'-0- 3'-end S/AS Methyl* "Stab 32" 2'-fluoro 2'-0- S/AS Methyl "Stab 33" 2'-fluoro 2'-deoxy* 5' and3'- - Usually S ends "Stab 34" 2'-fluoro 2'-0- 5' and 3'- Usually S Methyl* ends "Stab 35" 2'-fluoro*t 2'-0- Usually AS Methyl*t "Stab 36" 2'-fluoro*t 2'-0- Usually AS Methyl*t "Stab04H" 2'-fluoro$ Ribo$ 5' and 3'- 1 at 3'-end Ususally S ends "Stab06C" 2'-0-Methyll Ribo$ 5' and 3'- Ususally S ends "Stab07H" 2'-fluoro$ 2'-deoxy$ 5' and 3'- 1 at 3'-end Ususally S ends "Stab07mU" 2'-fluorol 2'-deoxyl 5' and 3'- Ususally S ends "StabO9H" Ribo$ Ribo$ 5' and 3'- 1 at 3'-end Ususally S ends "Stabl6C" Ribo$ 2'-0- 5' and 3'- Ususally S Methyll ends "Stabl6H" Ribo$ 2'-0- 5' and 3'- 1 at 3'-end Ususally S Methyll ends "Stabl8C" 2'-fluorol 2'-0- 5' and 3'- Ususally S Methyll ends "Stabl8H" 2'-fluoro$ 2'-0- 5' and 3'- 1 at 3'-end Ususally S Methyll ends "Stab52H" 2'-O-Methyl Ribo 5' and 3'- 1 at 3'-end Ususally S ends "Stab05C" 2'-fluoro Ribo Ususally AS "StabO5N" 2'-fluoro Ribo 1 at 3'-end Ususally AS "StablOC' Ribo Ribo Ususally AS "StablON" Ribo$ Ribo$ 1 at 3'-end Ususally AS "Stab35G*" 2'-fluorol 2'-0- Ususally AS Methyl "Stab35N*" 2'-fluorol 2'-0- 1 at 3'-end Ususally AS Methyl "Stab35rev*" 2'-O-Methyl 2'-fluoro Ususally AS "Stab50*" Ribo$ 2'-0- Ususally AS Methyll "Stab53*" I 2'-O-Methyl Ribo Ususally AS "Stab53N*" 2'-O-Methyl Ribo 1 at 3'-end Ususally AS Stab54 Ribo 2'-fluoro Ususally AS
CAP = any terminal cap, see for example Figure 6. All Stab chemistries can be used in combination with each other for duplexes of the invention (e.g., as combinations of sense and antisense strand chemistries), or alternately can be used in isolation, e.g., for single stranded nucleic acid molecules of the invention. All Stab chemistries can comprise 3'-overhang nucleotides having 2'--alkyl, 2'-deoxy-2'-fluoro, 2'-deoxy, LNA or other modified nucleotides or non-nucleotides. All Stab chemistries typically comprise about 19-21 nucleotides, but can vary as described herein. All Stab chemistries can also include a single ribonucleotide in the sense or passenger strand at the 1 1 th base paired position of the double-stranded nucleic acid duplex as determined from the '-end of the antisense or guide strand. All Stab chemistries can also have in place of the Stab designation above a 2'-deoxy-2'-fluoro modification at position 14 from the 5' end of the antisense strand regardless of whether it is a purine or pyrimidine at that position. All Stab chemistries of the antisense strand presented above can have a thymidine in place of a 2'-deoxy uridine at position 1, 2, and/or 3 from the 5' end of the antisense strand. S = sense strand. AS = antisense strand *Stab 23 has a single ribonucleotide adjacent to 3'-CAP. *Stab 24 and Stab 28 have a single ribonucleotide at 5'-terminus. *Stab 25, Stab 26, Stab 27, Stab 35, Stab 35G*, Stab 35N*, Stab 35rev*, Stab 36, Stab 50*, Stab53*, Stab 53N*, and Stab 54 have three ribonucleotides at 5'-terminus. *Stab 29, Stab 30, Stab 31, Stab 33, and Stab 34 any purine at first three nucleotide positions from 5'-terminus are ribonucleotides. p = phosphorothioate linkage. tStab 35 has 2'-O-methyl U at3'-overhangs and three ribonucleotides at 5'-terminus. tStab 36 has 2'-O-methyl overhangs that are complementary to the target sequence. (naturally occurring overhangs) and three ribonucleotides at 5'-terminus.
- Stab 04H, Stab 06C, Stabl07H, Stab07mU, Stab09H, Stabl6C, Stab 16H, Stabl8C, Stab 18H, Stab 52H, Stab 05C, Stab05N, StablOC, StablON, Stab35G*, Stab35N*, Stab35N*, Stab35rev*, Stab 50*, Stab 53*, Stab 53N*, Stab 54 have two 2'--methyl U3'-overhangs. Stab35G*, Stab 35N*, Stab35rev*, Stab50*, Stab53*, and Stab53N* do not allow for a 2'-0 methyl modification at position 14 of the guide strand as determined from the 5'-end.
Table 9
A. 2.5 pmol Synthesis Cycle ABI 394 Instrument Reagent Equivalents Amount Wait Time* Wait Time* 2'-O- Wait DNA methyl Time*RNA
Phosphorami 6.5 163 pL 45 sec 2.5 min 7.5 min dites S-Ethyl 23.8 238 pL 45 sec 2.5 min 7.5 min Tetrazole Acetic 100 233 pL 5 sec 5 sec 5 sec Anhydride N-HBVhyl 186 233 pL 5 sec 5 sec 5 sec Imidazole TCA 176 2.3 mL 21 sec 21 sec 21 sec Iodine 11.2 1.7 mL 45 sec 45 sec 45 sec Beaucage 12.9 645 pL 100 sec 300 sec 300 sec Acetonitrile NA 6.67 mL NA NA NA B. 0.2 pmol Synthesis Cycle ABI 394 Instrument Reagent Equivalents Amount Wait Time* Wait Time* 2'-O- Wait DNA methyl Time*RNA
Phosphorami 15 31 pL 45 sec 233 sec 465 sec dites S-Ethyl 38.7 31 pL 45 sec 233 min 465 sec Tetrazole Acetic 655 124 pL 5 sec 5 sec 5 sec Anhydride N-HBVhyl 1245 124 pL 5 sec 5 sec 5 sec Imidazole TCA 700 732 pL 10 sec 10 sec 10 sec Iodine 20.6 244 pL 15 sec 15 sec 15 sec Beaucage 7.7 232 pL 100 sec 300 sec 300 sec Acetonitrile NA 2.64 mL NA NA NA
C. 0.2 pmol Synthesis Cycle 96 well Instrument Reagent Equivalents: Amount: Wait Time* Wait Time* Wait Time* DNA/2'-O- DNA/2'-O- DNA 2'-O-methyl Ribo methyl/Ribo methyl/Ribo
Phosphorami 22/33/66 40/60/120 pL 60 sec 180 sec 360sec dites S-Ethyl 70/105/210 40/60/120 pL 60 sec 180 min 360 sec Tetrazole Acetic 265/265/265 50/50/50 pL 10 sec 10 sec 10 sec Anhydride N-HBVhyl 502/502/502 50/50/50 pL 10 sec 10 sec 10 sec Imidazole TCA 238/475/475 250/500/500 pL 15 sec 15 sec 15 sec Iodine 6.8/6.8/6.8 80/80/80 pL 30 sec 30 sec 30 sec Beaucage 34/51/51 80/120/120 100 sec 200 sec 200 sec Acetonitrile NA 1150/1150/1150 NA NA NA pL • Wait time does not include contact time during delivery.
• Tandem synthesis utilizes double coupling of linker molecule
Table 10: Cmposition of Select Lipid Nanoparticle Formulations LNP Lipid Components and Molar Ratios siNA N/P Identifier Duplex LNP-1 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380648 (50%) 00E LNP-2 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380783 (50%) OOOR LNP-3 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380665 (50%) 00N LNP-4 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380645 (50%) OOOD LNP-5 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380408 (50%) OOOR LNP-6 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380633 (50%) 00OU LNP-7 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380767 (50%) OOOR LNP-8 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380730 (50%) 000C LNP-9 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380777 (50%) 000H LNP-10 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380740 (50%) 000V LNP-ll Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380558 (50%) 000M LNP-12 Compound Cholesterol DSPC PEG-DMG R-008380406- 6 32 (30%) (10%) (2%) OOOY
(50%)6 LNP-13 Compou6nd Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380764 (50%) OOOP LNP-14 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380772 (50%) OOOP LNP-15 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380389 (50%) OOOD LNP-16 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380362 (50%) 000H LNP-17 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380363 (50%) 000S LNP-18 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380340 (50%) OOOF LNP-19 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380689 (50%) 000H LNP-20 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380756 (50%) OOOP LNP-21 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380736 (50%) 00E LNP-22 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380757 (50%) OOOY LNP-23 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380753 (50%) 00N LNP-24 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380737 (50%) 00N LNP-25 Compound Cholesterol DSPC PEG-DMG R-008380734- 6 000M
32 (30%) (10%) (2%) (50%) LNP-26 Compound Cholesterol DSPC PEG-DMG 6 32 (30%) (10%) (2%) R-008380791 (50%) OOOR N/P ratio = Nitrogen:Phosphorous ratio between cationic lipid and nucleic acid
Table 11: ChemicalStructures ofLipids in Formulationsof Table 10
Lipid ChemicalStructure Compound N 32
Cholesterol
DSPC 0 o
12 0
% 0N' O,, ON y 0 012 HH
2K 12'
SIRIFD00001WOPCTSEQ 13 Nov 2019
<110> Steve Bartz Duncan Brown Michael Robinson
<120> RNA Interference Mediated Inhibition of Hepatitis B Virus (HBV) Gene Expression using Short Interfering Nucleic Acid (siNA)
<130> SIR-IFD-00001 2019264591
<160> 526
<170> PatentIn version 3.5
<210> 1 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 1 ucguggugga cuucucuca 19
<210> 2 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 2 gugguggacu ucucucaau 19
<210> 3 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 3 gccgauccau acugcggaa 19
<210> 4 Page 1
SIRIFD00001WOPCTSEQ 13 Nov 2019
<211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 4 ccgauccaua cugcggaac 19 2019264591
<210> 5 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 5 cauccugcug cuaugccuc 19
<210> 6 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 6 ugcugcuaug ccucaucuu 19
<210> 7 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 7 gguggacuuc ucucaauuu 19
<210> 8 <211> 19 <212> RNA <213> Artificial Sequence
Page 2
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<400> 8 ugguggacuu cucucaauu 19
<210> 9 <211> 19 <212> RNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<400> 9 uagacucgug guggacuuc 19
<210> 10 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 10 uccucugccg auccauacu 19
<210> 11 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 11 ugccgaucca uacugcgga 19
<210> 12 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 12 Page 3
SIRIFD00001WOPCTSEQ 13 Nov 2019
uggauguguc ugcggcguu 19
<210> 13 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<400> 13 cgauccauac ugcggaacu 19
<210> 14 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 14 cgcaccucuc uuuacgcgg 19
<210> 15 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 15 cugccgaucc auacugcgg 19
<210> 16 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 16 cgugguggac uucucucaa 19
<210> 17 Page 4
SIRIFD00001WOPCTSEQ 13 Nov 2019
<211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 17 cugcugcuau gccucaucu 19 2019264591
<210> 18 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 18 ccugcugcua ugccucauc 19
<210> 19 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 19 cuagacucgu gguggacuu 19
<210> 20 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 20 uccugcugcu augccucau 19
<210> 21 <211> 19 <212> RNA <213> Artificial Sequence
Page 5
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<400> 21 gacucguggu ggacuucuc 19
<210> 22 <211> 19 <212> RNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<400> 22 auccauacug cggaacucc 19
<210> 23 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 23 cucugccgau ccauacugc 19
<210> 24 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 24 gauccauacu gcggaacuc 19
<210> 25 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 25 Page 6
SIRIFD00001WOPCTSEQ 13 Nov 2019
gaagaacucc cucgccucg 19
<210> 26 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<400> 26 aagccuccaa gcugugccu 19
<210> 27 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 27 agaagaacuc ccucgccuc 19
<210> 28 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 28 ggagugugga uucgcacuc 19
<210> 29 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 29 ccucugccga uccauacug 19
<210> 30 Page 7
SIRIFD00001WOPCTSEQ 13 Nov 2019
<211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 30 caagccucca agcugugcc 19 2019264591
<210> 31 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 31 uccauacugc ggaacuccu 19
<210> 32 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 32 cagagucuag acucguggu 19
<210> 33 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 33 aagaagaacu cccucgccu 19
<210> 34 <211> 19 <212> RNA <213> Artificial Sequence
Page 8
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<400> 34 gaguguggau ucgcacucc 19
<210> 35 <211> 19 <212> RNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<400> 35 ucuagacucg ugguggacu 19
<210> 36 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 36 gcugcuaugc cucaucuuc 19
<210> 37 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 37 agucuagacu cguggugga 19
<210> 38 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 38 Page 9
SIRIFD00001WOPCTSEQ 13 Nov 2019
cuccucugcc gauccauac 19
<210> 39 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<400> 39 uggcucaguu uacuagugc 19
<210> 40 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 40 gucuagacuc gugguggac 19
<210> 41 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 41 uucaagccuc caagcugug 19
<210> 42 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 42 cuaugggagu gggccucag 19
<210> 43 Page 10
SIRIFD00001WOPCTSEQ 13 Nov 2019
<211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 43 cucguggugg acuucucuc 19 2019264591
<210> 44 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 44 ccuaugggag ugggccuca 19
<210> 45 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 45 aagaacuccc ucgccucgc 19
<210> 46 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 46 ucugccgauc cauacugcg 19
<210> 47 <211> 19 <212> RNA <213> Artificial Sequence
Page 11
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<400> 47 agagucuaga cucguggug 19
<210> 48 <211> 19 <212> RNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<400> 48 gaagaagaac ucccucgcc 19
<210> 49 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 49 ucaagccucc aagcugugc 19
<210> 50 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 50 agccuccaag cugugccuu 19
<210> 51 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 51 Page 12
SIRIFD00001WOPCTSEQ 13 Nov 2019
agacucgugg uggacuucu 19
<210> 52 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 52 ucguggugga cuucucucat t 21
<210> 53 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 53 ugagagaagu ccaccacgau u 21
<210> 54 <211> 21 <212> DNA <213> Artificial Sequence Page 13
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence 2019264591
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy
<220> <221> misc_feature Page 14
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 54 gugguggacu ucucucaaut t 21 2019264591
<210> 55 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 15
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (20)..(21) <223> 2'-O-methyl
<400> 55 auugagagaa guccaccacu u 21
<210> 56 <211> 21 <212> DNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 56 gccgauccau acugcggaat t 21
<210> 57 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 57 uuccgcagua uggaucggcu u 21
Page 16
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 58 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature 2019264591
<222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 58 gugguggacu ucucucaaut t 21
<210> 59 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 59 auugagagaa guccaccacu u 21
<210> 60 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic Page 17
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) 2019264591
<223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
Page 18
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(18) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 60 ccgauccaua cugcggaact t 21
<210> 61 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(9) <223> 2'-O-methyl
Page 19
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<400> 61 guuccgcagu auggaucggu u 21
<210> 62 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence Page 20
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap 2019264591
<400> 62 cauccugcug cuaugccuct t 21
<210> 63 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 63 gaggcauagc agcaggaugu u 21
<210> 64 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> Page 21
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 64 ugcugcuaug ccucaucuut t 21
<210> 65 <211> 21 2019264591
<212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 65 aagaugaggc auagcagcau u 21
<210> 66 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(3) Page 22
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(14) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 66 gguggacuuc ucucaauuut t 21
<210> 67 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(16) Page 23
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(19) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 67 aaauugagag aaguccaccu u 21
<210> 68 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy
<220> <221> misc_feature Page 24
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(7) <223> 2'-deoxy
<220> <221> misc_feature 2019264591
<222> (8)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 68 ugguggacuu cucucaauut t 21
<210> 69 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(12) <223> 2'-O-methyl
<220> <221> misc_feature Page 25
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (13)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (17)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<400> 69 aauugagaga aguccaccau u 21
<210> 70 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 70 uagacucgug guggacuuct t 21
<210> 71 <211> 21 Page 26
SIRIFD00001WOPCTSEQ 13 Nov 2019
<212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl 2019264591
<400> 71 gaaguccacc acgagucuau u 21
<210> 72 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 72 uccucugccg auccauacut t 21
<210> 73 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> Page 27
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 73 aguauggauc ggcagaggau u 21
<210> 74 <211> 21 2019264591
<212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
Page 28
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (10)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (13)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 74 uagacucgug guggacuuct t 21
<210> 75 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
Page 29
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<400> 75 gaaguccacc acgagucuau u 21
<210> 76 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence Page 30
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap 2019264591
<400> 76 ugccgaucca uacugcggat t 21
<210> 77 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 77 uccgcaguau ggaucggcau u 21
<210> 78 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> Page 31
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 78 uggauguguc ugcggcguut t 21
<210> 79 <211> 21 2019264591
<212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 79 aacgccgcag acacauccau u 21
<210> 80 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 80 cgauccauac ugcggaacut t 21
Page 32
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 81 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> 2019264591
<221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 81 aguuccgcag uauggaucgu u 21
<210> 82 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 82 cgcaccucuc uuuacgcggt t 21
<210> 83 <211> 21 <212> RNA <213> Artificial Sequence
<220> Page 33
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 83 ccgcguaaag agaggugcgu u 21 2019264591
<210> 84 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 84 cugccgaucc auacugcggt t 21
<210> 85 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
Page 34
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 85 ccgcaguaug gaucggcagu u 21
<210> 86 <211> 21 <212> DNA <213> Artificial Sequence
<220> 2019264591
<223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy Page 35
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (9)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 86 uggauguguc ugcggcguut t 21
<210> 87 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic Page 36
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(18) <223> 2'-deoxy-2'-fluoro
<220> Page 37
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<400> 87 aacgccgcag acacauccau u 21
<210> 88 <211> 21 2019264591
<212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 88 ugguggacuu cucucaauut t 21
<210> 89 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 89 aauugagaga aguccaccau u 21
Page 38
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 90 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> 2019264591
<221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 90 cgugguggac uucucucaat t 21
<210> 91 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 91 uugagagaag uccaccacgu u 21
<210> 92 <211> 21 <212> DNA <213> Artificial Sequence
<220> Page 39
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature 2019264591
<222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 92 cugcugcuau gccucaucut t 21
<210> 93 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 93 agaugaggca uagcagcagu u 21
<210> 94 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence Page 40
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap 2019264591
<400> 94 ccugcugcua ugccucauct t 21
<210> 95 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 95 gaugaggcau agcagcaggu u 21
<210> 96 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> Page 41
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 96 gguggacuuc ucucaauuut t 21
<210> 97 <211> 21 2019264591
<212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 97 aaauugagag aaguccaccu u 21
<210> 98 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(5) Page 42
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) 2019264591
<223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(16) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 98 cuagacucgu gguggacuut t 21
<210> 99 <211> 21 <212> RNA <213> Artificial Sequence
<220> Page 43
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> Synthetic
<220> <221> misc_feature <222> (4)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<400> 99 aaguccacca cgagucuagu u 21
<210> 100 <211> 21 <212> DNA <213> Artificial Sequence
<220> Page 44
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature 2019264591
<222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 100 uccugcugcu augccucaut t 21
<210> 101 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 101 augaggcaua gcagcaggau u 21
<210> 102 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence Page 45
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy-2'-fluoro
<220> Page 46
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (17)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 102 ugccgaucca uacugcggat t 21
<210> 103 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> Page 47
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (11)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<400> 103 uccgcaguau ggaucggcau u 21
<210> 104 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
Page 48
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap 2019264591
<400> 104 cuagacucgu gguggacuut t 21
<210> 105 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 105 aaguccacca cgagucuagu u 21
<210> 106 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature Page 49
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (21)..(21) <223> inverted abasic cap
<400> 106 gacucguggu ggacuucuct t 21
<210> 107 <211> 21 <212> RNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 107 gagaagucca ccacgagucu u 21
<210> 108 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (2)..(4) <223> 2'-deoxy-2'-fluoro Page 50
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 108 auccauacug cggaacucct t 21
<210> 109 <211> 21 Page 51
SIRIFD00001WOPCTSEQ 13 Nov 2019
<212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (5)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> Page 52
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 109 ggaguuccgc aguauggauu u 21
<210> 110 <211> 21 2019264591
<212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(12) <223> 2'-deoxy-2'-fluoro
Page 53
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 110 cucugccgau ccauacugct t 21
<210> 111 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
Page 54
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(21) <223> 2'-O-methyl
<400> 111 gcaguaugga ucggcagagu u 21
<210> 112 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
Page 55
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap 2019264591
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 112 gauccauacu gcggaacuct t 21
<210> 113 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 113 gaguuccgca guauggaucu u 21
<210> 114 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> Page 56
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 114 2019264591
gaagaacucc cucgccucgt t 21
<210> 115 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 115 cgaggcgagg gaguucuucu u 21
<210> 116 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(3) Page 57
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(11) 2019264591
<223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 116 aagccuccaa gcugugccut t 21
<210> 117 <211> 21 <212> RNA <213> Artificial Sequence
<220> Page 58
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 117 aggcacagcu uggaggcuuu u 21
<210> 118 Page 59
SIRIFD00001WOPCTSEQ 13 Nov 2019
<211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) 2019264591
<223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 118 agaagaacuc ccucgccuct t 21
<210> 119 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 119 gaggcgaggg aguucuucuu u 21
<210> 120 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
Page 60
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap 2019264591
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy
<220> Page 61
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(17) <223> 2'-deoxy
<220> 2019264591
<221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 120 cgauccauac ugcggaacut t 21
<210> 121 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(10) <223> 2'-O-methyl
<220> Page 62
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<400> 121 aguuccgcag uauggaucgu u 21
<210> 122 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
Page 63
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 122 ggagugugga uucgcacuct t 21
<210> 123 2019264591
<211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 123 gagugcgaau ccacacuccu u 21
<210> 124 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 64
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (6)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature 2019264591
<222> (9)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 124 ccucugccga uccauacugt t 21
Page 65
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 125 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature 2019264591
<222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(21) <223> 2'-O-methyl
<400> 125 caguauggau cggcagaggu u 21
Page 66
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 126 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature 2019264591
<222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) Page 67
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(19) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 126 caagccucca agcugugcct t 21
<210> 127 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(13) Page 68
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(18) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<400> 127 ggcacagcuu ggaggcuugu u 21
<210> 128 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 128 uccauacugc ggaacuccut t 21
<210> 129 <211> 21 <212> RNA Page 69
SIRIFD00001WOPCTSEQ 13 Nov 2019
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl 2019264591
<400> 129 aggaguuccg caguauggau u 21
<210> 130 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 130 cagagucuag acucguggut t 21
<210> 131 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature Page 70
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (20)..(21) <223> 2'-O-methyl
<400> 131 accacgaguc uagacucugu u 21
<210> 132 <211> 21 <212> DNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 132 Page 71
SIRIFD00001WOPCTSEQ 13 Nov 2019
aagaagaacu cccucgccut t 21
<210> 133 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 133 aggcgaggga guucuucuuu u 21
<210> 134 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) Page 72
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 134 gaguguggau ucgcacucct t 21 2019264591
<210> 135 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 135 ggagugcgaa uccacacucu u 21
<210> 136 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
Page 73
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 136 ucuagacucg ugguggacut t 21
<210> 137 <211> 21 <212> RNA <213> Artificial Sequence
<220> 2019264591
<223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 137 aguccaccac gagucuagau u 21
<210> 138 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy
<220> <221> misc_feature Page 74
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (8)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(11) <223> 2'-deoxy
<220> <221> misc_feature 2019264591
<222> (12)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 138 uccucugccg auccauacut t 21
<210> 139 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature Page 75
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature 2019264591
<222> (6)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(21) <223> 2'-O-methyl
<400> 139 aguauggauc ggcagaggau u 21
<210> 140 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> Page 76
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 140 2019264591
caagccucca agcugugcct t 21
<210> 141 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 141 ggcacagcuu ggaggcuugu u 21
<210> 142 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(3) Page 77
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(6) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
Page 78
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 142 ccugcugcua ugccucauct t 21
<210> 143 2019264591
<211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
Page 79
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(21) <223> 2'-O-methyl 2019264591
<400> 143 gaugaggcau agcagcaggu u 21
<210> 144 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 144 cucugccgau ccauacugct t 21
<210> 145 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature Page 80
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (20)..(21) <223> 2'-O-methyl
<400> 145 gcaguaugga ucggcagagu u 21
<210> 146 <211> 21 <212> DNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(14) <223> 2'-deoxy-2'-fluoro
<220> Page 81
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (17)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 146 cagagucuag acucguggut t 21
<210> 147 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(8) <223> 2'-O-methyl
<220> Page 82
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (9)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (15)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<400> 147 accacgaguc uagacucugu u 21
<210> 148 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy-2'-fluoro
Page 83
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 148 gcugcuaugc cucaucuuct t 21
<210> 149 <211> 21 <212> RNA Page 84
SIRIFD00001WOPCTSEQ 13 Nov 2019
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(5) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(18) <223> 2'-O-methyl
<220> <221> misc_feature Page 85
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 149 gaagaugagg cauagcagcu u 21 2019264591
<210> 150 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 150 ccucugccga uccauacugt t 21
<210> 151 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl Page 86
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 151 caguauggau cggcagaggu u 21
<210> 152 <211> 21 <212> DNA <213> Artificial Sequence 2019264591
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) Page 87
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(17) 2019264591
<223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 152 ucuagacucg ugguggacut t 21
<210> 153 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) Page 88
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(13) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(21) <223> 2'-O-methyl
<400> 153 aguccaccac gagucuagau u 21
<210> 154 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 89
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (2)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature 2019264591
<222> (7)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap Page 90
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 154 cauccugcug cuaugccuct t 21
<210> 155 <211> 21 <212> RNA <213> Artificial Sequence 2019264591
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro Page 91
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (15)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<400> 155 gaggcauagc agcaggaugu u 21
<210> 156 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(8) Page 92
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) 2019264591
<223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 156 agucuagacu cgugguggat t 21
<210> 157 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) Page 93
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 157 uccaccacga gucuagacuu u 21
<210> 158 <211> 21 <212> DNA <213> Artificial Sequence
<220> Page 94
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature 2019264591
<222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro Page 95
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(19) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 158 ggagugugga uucgcacuct t 21
<210> 159 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(12) <223> 2'-deoxy-2'-fluoro Page 96
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 159 gagugcgaau ccacacuccu u 21
<210> 160 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) Page 97
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(4) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(19) <223> 2'-deoxy-2'-fluoro
Page 98
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 160 ugcugcuaug ccucaucuut t 21
<210> 161 2019264591
<211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
Page 99
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(17) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<400> 161 aagaugaggc auagcagcau u 21
<210> 162 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy Page 100
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 162 uccauacugc ggaacuccut t 21
<210> 163 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic Page 101
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (4)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(9) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(21) <223> 2'-O-methyl
<400> 163 aggaguuccg caguauggau u 21
<210> 164 <211> 21 Page 102
SIRIFD00001WOPCTSEQ 13 Nov 2019
<212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this 2019264591
sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 164 cuccucugcc gauccauact t 21
<210> 165 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 165 guauggaucg gcagaggagu u 21
<210> 166 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> Page 103
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap 2019264591
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 104
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (13)..(16) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature 2019264591
<222> (21)..(21) <223> inverted abasic cap
<400> 166 gauccauacu gcggaacuct t 21
<210> 167 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 105
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature 2019264591
<222> (15)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 167 gaguuccgca guauggaucu u 21
<210> 168 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(6) <223> 2'-deoxy
<220> Page 106
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (7)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> 2019264591
<221> misc_feature <222> (15)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 168 gaagaacucc cucgccucgt t 21
<210> 169 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(13) <223> 2'-O-methyl
<220> Page 107
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (14)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 169 2019264591
cgaggcgagg gaguucuucu u 21
<210> 170 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy
Page 108
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(10) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (11)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 170 ucguggugga cuucucucat t 21
<210> 171 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
Page 109
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (14)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<400> 171 ugagagaagu ccaccacgau u 21
<210> 172 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy Page 110
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (6)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 172 Page 111
SIRIFD00001WOPCTSEQ 13 Nov 2019
uccugcugcu augccucaut t 21
<210> 173 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<220> <221> misc_feature <222> (4)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> Page 112
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (16)..(21) <223> 2'-O-methyl
<400> 173 augaggcaua gcagcaggau u 21
<210> 174 <211> 21 2019264591
<212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 174 auccauacug cggaacucct t 21
<210> 175 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 175 ggaguuccgc aguauggauu u 21
Page 113
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 176 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> 2019264591
<221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 176 uggcucaguu uacuagugct t 21
<210> 177 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 177 gcacuaguaa acugagccau u 21
<210> 178 <211> 21 <212> DNA <213> Artificial Sequence
<220> Page 114
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature 2019264591
<222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 178 aagccuccaa gcugugccut t 21
<210> 179 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 179 aggcacagcu uggaggcuuu u 21
<210> 180 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence Page 115
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(7) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (8)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 180 agaagaacuc ccucgccuct t 21
<210> 181 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro Page 116
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (6)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(19) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 181 gaggcgaggg aguucuucuu u 21
<210> 182 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (2)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(7) Page 117
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) 2019264591
<223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 182 gucuagacuc gugguggact t 21
<210> 183 <211> 21 <212> RNA <213> Artificial Sequence
<220> Page 118
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
Page 119
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 183 guccaccacg agucuagacu u 21
<210> 184 <211> 21 <212> DNA <213> Artificial Sequence
<220> 2019264591
<223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy Page 120
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (12)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 184 cugcugcuau gccucaucut t 21
<210> 185 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(8) <223> 2'-O-methyl Page 121
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<400> 185 agaugaggca uagcagcagu u 21
<210> 186 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic Page 122
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) 2019264591
<223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
Page 123
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (14)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 186 cugccgaucc auacugcggt t 21
<210> 187 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
Page 124
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (10)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<400> 187 ccgcaguaug gaucggcagu u 21
<210> 188 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence Page 125
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(3) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (4)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 188 Page 126
SIRIFD00001WOPCTSEQ 13 Nov 2019
uucaagccuc caagcugugt t 21
<210> 189 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<220> <221> misc_feature <222> (4)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(21) <223> 2'-O-methyl
<400> 189 cacagcuugg aggcuugaau u 21
<210> 190 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this Page 127
SIRIFD00001WOPCTSEQ 13 Nov 2019
sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) 2019264591
<223> inverted abasic cap
<400> 190 agucuagacu cgugguggat t 21
<210> 191 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 191 uccaccacga gucuagacuu u 21
<210> 192 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
Page 128
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 192 gucuagacuc gugguggact t 21
<210> 193 2019264591
<211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 193 guccaccacg agucuagacu u 21
<210> 194 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 194 cuaugggagu gggccucagt t 21 Page 129
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 195 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 195 cugaggccca cucccauagu u 21
<210> 196 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 196 uucaagccuc caagcugugt t 21
<210> 197 <211> 21 <212> RNA <213> Artificial Sequence
Page 130
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 197 cacagcuugg aggcuugaau u 21 2019264591
<210> 198 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> Page 131
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (8)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (11)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 198 gacucguggu ggacuucuct t 21
<210> 199 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> Page 132
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (11)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 199 gagaagucca ccacgagucu u 21
<210> 200 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
Page 133
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 200 gcugcuaugc cucaucuuct t 21
<210> 201 2019264591
<211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 201 gaagaugagg cauagcagcu u 21
<210> 202 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 134
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (2)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature 2019264591
<222> (4)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 202 cgugguggac uucucucaat t 21
<210> 203 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature Page 135
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (4)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature 2019264591
<222> (15)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<400> 203 uugagagaag uccaccacgu u 21
<210> 204 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> Page 136
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy
<220> 2019264591
<221> misc_feature <222> (4)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(19) Page 137
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 204 uggcucaguu uacuagugct t 21 2019264591
<210> 205 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(18) Page 138
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<400> 205 gcacuaguaa acugagccau u 21 2019264591
<210> 206 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(12) <223> 2'-deoxy
<220> <221> misc_feature Page 139
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (13)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy
<220> <221> misc_feature 2019264591
<222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 206 cuccucugcc gauccauact t 21
<210> 207 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(7) <223> 2'-O-methyl
<220> <221> misc_feature Page 140
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (8)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(11) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(21) <223> 2'-O-methyl
<400> 207 guauggaucg gcagaggagu u 21
<210> 208 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(3) <223> 2'-deoxy
<220> Page 141
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy
<220> 2019264591
<221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 208 gaguguggau ucgcacucct t 21
Page 142
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 209 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> 2019264591
<221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(19) Page 143
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 209 ggagugcgaa uccacacucu u 21 2019264591
<210> 210 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy
<220> <221> misc_feature Page 144
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (5)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> <221> misc_feature 2019264591
<222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 210 cgcaccucuc uuuacgcggt t 21
<210> 211 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 145
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature 2019264591
<222> (7)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<400> 211 ccgcguaaag agaggugcgu u 21
<210> 212 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature Page 146
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> 2019264591
<221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 212 ccgauccaua cugcggaact t 21
<210> 213 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 213 guuccgcagu auggaucggu u 21
<210> 214 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature <222> (1)..(1) Page 147
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(3) 2019264591
<223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 214 cuaugggagu gggccucagt t 21
<210> 215 Page 148
SIRIFD00001WOPCTSEQ 13 Nov 2019
<211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(6) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<400> 215 cugaggccca cucccauagu u 21
<210> 216 <211> 21 <212> DNA <213> Artificial Sequence
<220> Page 149
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence
<220> <221> misc_feature 2019264591
<222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 216 aagaagaacu cccucgccut t 21
<210> 217 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 217 aggcgaggga guucuucuuu u 21
<210> 218 <211> 21 <212> DNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(19) <223> ribonucleotide unmodified or modified as described for this sequence Page 150
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> Page 151
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(19) <223> 2'-deoxy
<220> 2019264591
<221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 218 gccgauccau acugcggaat t 21
<210> 219 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> Page 152
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<400> 219 uuccgcagua uggaucggcu u 21
<210> 220 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> Page 153
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(4) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (5)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) Page 154
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) 2019264591
<223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 220 ccgauccaua cugcggaacu u 21
<210> 221 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) Page 155
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) 2019264591
<223> phosphorothioate linkage
<400> 221 guuccgcagu auggaucggu u 21
<210> 222 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (3)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(18) Page 156
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) 2019264591
<223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 222 ccgauccaua cugcggaacu u 21
<210> 223 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (2)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(17) Page 157
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) 2019264591
<223> phosphorothioate linkage
<400> 223 guuccgcagu auggaucggu u 21
<210> 224 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) Page 158
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 224 gccgauccau acugcggaau u 21
<210> 225 <211> 21 <212> RNA <213> Artificial Sequence
<220> Page 159
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy-2'-fluoro
Page 160
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage 2019264591
<400> 225 uuccgcagua uggaucggcu u 21
<210> 226 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (2)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
Page 161
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage 2019264591
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 226 gccgauccau acugcggaau u 21
<210> 227 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy-2'-fluoro
Page 162
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage 2019264591
<400> 227 uuccgcagua uggaucggcu u 21
<210> 228 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
Page 163
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage 2019264591
<400> 228 uuccgcagua uggaucggcu u 21
<210> 229 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
Page 164
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (12)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 229 gccgauccau acugcggaau u 21
<210> 230 <211> 21 <212> RNA Page 165
SIRIFD00001WOPCTSEQ 13 Nov 2019
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 166
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (17)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature 2019264591
<222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 230 uuccgcagua uggaucggcu u 21
<210> 231 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 231 gccgauccau acugcggaau u 21 Page 167
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 232 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 232 uuccgcagua uggaucggcu u 21
<210> 233 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(4) <223> 2'-O-methyl
Page 168
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(8) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (9)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 233 gugguggacu ucucucaauu u 21
<210> 234 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
Page 169
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(14) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 234 auugagagaa guccaccacu u 21
<210> 235 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
Page 170
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap 2019264591
<400> 235 gugguggacu ucucucaauu u 21
<210> 236 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 236 auugagagaa guccaccacu u 21
<210> 237 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) Page 171
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(4) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 237 ucguggugga cuucucucau u 21
<210> 238 Page 172
SIRIFD00001WOPCTSEQ 13 Nov 2019
<211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(9) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
Page 173
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 238 ugagagaagu ccaccacgau u 21
<210> 239 <211> 21 <212> RNA <213> Artificial Sequence
<220> 2019264591
<223> Synthetic
<220> <221> misc_feature <222> (4)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
Page 174
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 239 ugagagaagu ccaccacgau u 21
<210> 240 <211> 21 <212> RNA <213> Artificial Sequence
<220> 2019264591
<223> Synthetic
<220> <221> misc_feature <222> (10)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 240 ugagagaagu ccaccacgau u 21
<210> 241 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) Page 175
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(4) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 241 ucguggugga cuucucucau u 21
<210> 242 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(9) Page 176
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 242 ugagagaagu ccaccacgau u 21
<210> 243 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(15) Page 177
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 243 ugagagaagu ccaccacgau u 21
<210> 244 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(21) Page 178
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) 2019264591
<223> inverted abasic cap
<400> 244 ucguggugga cuucucucau u 21
<210> 245 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(9) Page 179
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(12) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 245 ccgauccaua cugcggaacu u 21
<210> 246 Page 180
SIRIFD00001WOPCTSEQ 13 Nov 2019
<211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(5) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
Page 181
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 246 guuccgcagu auggaucggu u 21
<210> 247 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
Page 182
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (14)..(14) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(21) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 247 gccgauccau acugcggaau u 21
<210> 248 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
Page 183
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (17)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 248 uuccgcagua uggaucggcu u 21
<210> 249 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-deoxy-2'-fluoro
Page 184
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 249 gccgauccau acugcggaau u 21
<210> 250 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(16) <223> 2'-O-methyl
Page 185
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage 2019264591
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 250 gugguggacu ucucucaauu u 21
<210> 251 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
Page 186
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 251 auugagagaa guccaccacu u 21
<210> 252 <211> 21 <212> RNA <213> Artificial Sequence
<220> 2019264591
<223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
Page 187
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 252 gugguggacu ucucucaauu u 21
<210> 253 <211> 21 <212> RNA <213> Artificial Sequence
<220> 2019264591
<223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
Page 188
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 253 ucguggugga cuucucucau u 21
<210> 254 <211> 21 <212> RNA <213> Artificial Sequence
<220> 2019264591
<223> Synthetic
<220> <221> misc_feature <222> (4)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 254 guuccgcagu auggaucggu u 21
<210> 255 Page 189
SIRIFD00001WOPCTSEQ 13 Nov 2019
<211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (6)..(6) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 255 guuccgcagu auggaucggu u 21
<210> 256 <211> 21 <212> RNA <213> Artificial Sequence
<220> Page 190
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> Synthetic
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(17) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 256 auugagagaa guccaccacu u 21
<210> 257 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(8) Page 191
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 257 gugguggacu ucucucaauu u 21
<210> 258 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(4) Page 192
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(8) 2019264591
<223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 258 gugguggacu ucucucaauu u 21
<210> 259 <211> 21 <212> RNA <213> Artificial Sequence
<220> Page 193
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> Synthetic
<220> <221> misc_feature <222> (4)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(14) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 259 auugagagaa guccaccacu u 21
<210> 260 <211> 21 <212> RNA <213> Artificial Sequence
<220> Page 194
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> Synthetic
<220> <221> misc_feature <222> (4)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 260 ugagagaagu ccaccacgau u 21
<210> 261 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(7) Page 195
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(12) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 261 ccgauccaua cugcggaacu u 21
<210> 262 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) Page 196
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 262 ccgauccaua cugcggaacu u 21
<210> 263 <211> 21 <212> RNA <213> Artificial Sequence
<220> Page 197
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> Synthetic
<220> <221> misc_feature <222> (4)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 263 auugagagaa guccaccacu u 21
<210> 264 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) Page 198
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> inverted abasic cap
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) 2019264591
<223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 264 ucguggugga cuucucucau u 21
<210> 265 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 265 ugagagaagu ccaccacgau u 21
<210> 266 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> Page 199
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (4)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) Page 200
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 266 guuccgcagu auggaucggu u 21 2019264591
<210> 267 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) Page 201
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> phosphorothioate linkage
<400> 267 uuccgcagua uggaucggcu u 21
<210> 268 <211> 21 <212> RNA <213> Artificial Sequence 2019264591
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 268 auugagagaa guccaccacu u 21
<210> 269 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> Page 202
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) Page 203
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 269 ucguggugga cuucucucau u 21 2019264591
<210> 270 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 270 ccgauccaua cugcggaacu u 21
<210> 271 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> Page 204
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 271 2019264591
guuccgcagu auggaucggu u 21
<210> 272 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> Page 205
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (11)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 272 ccgauccaua cugcggaacu u 21
<210> 273 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> Page 206
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (1)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) Page 207
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 273 guuccgcagu auggaucggu u 21 2019264591
<210> 274 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(8) Page 208
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(12) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
Page 209
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 274 ccgauccaua cugcggaacu u 21
<210> 275 2019264591
<211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
Page 210
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 275 guuccgcagu auggaucggu u 21
<210> 276 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
Page 211
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 276 guuccgcagu auggaucggu u 21
<210> 277 <211> 21 <212> RNA Page 212
SIRIFD00001WOPCTSEQ 13 Nov 2019
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap 2019264591
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> <221> misc_feature Page 213
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy
<220> <221> misc_feature 2019264591
<222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 277 ccgauccaua cugcggaacu u 21
<210> 278 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature Page 214
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (3)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(6) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage Page 215
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 278 uuccgcagua uggaucggcu u 21
<210> 279 <211> 21 <212> RNA <213> Artificial Sequence 2019264591
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl Page 216
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(15) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 279 gccgauccau acugcggaau u 21
<210> 280 <211> 21 Page 217
SIRIFD00001WOPCTSEQ 13 Nov 2019
<212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap 2019264591
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(11) <223> 2'-deoxy
<220> Page 218
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (14)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 280 gugguggacu ucucucaauu u 21
<210> 281 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> Page 219
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (2)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) Page 220
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> phosphorothioate linkage
<400> 281 auugagagaa guccaccacu u 21
<210> 282 <211> 21 <212> RNA <213> Artificial Sequence 2019264591
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) Page 221
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 282 auugagagaa guccaccacu u 21
<210> 283 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(4) Page 222
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
Page 223
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 283 gugguggacu ucucucaauu u 21
<210> 284 <211> 21 <212> RNA <213> Artificial Sequence
<220> 2019264591
<223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
Page 224
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(16) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 284 gugguggacu ucucucaauu u 21
<210> 285 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(2) <223> 2'-O-methyl
Page 225
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (3)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature Page 226
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (20)..(21) <223> phosphorothioate linkage
<400> 285 auugagagaa guccaccacu u 21
<210> 286 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(7) <223> 2'-deoxy
<220> <221> misc_feature Page 227
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (8)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 286 ucguggugga cuucucucau u 21
Page 228
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 287 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature 2019264591
<222> (1)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro Page 229
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 287 ugagagaagu ccaccacgau u 21
<210> 288 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro Page 230
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 288 guuccgcagu auggaucggu u 21
<210> 289 <211> 21 Page 231
SIRIFD00001WOPCTSEQ 13 Nov 2019
<212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap 2019264591
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> Page 232
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 289 ccgauccaua cugcggaacu u 21
<210> 290 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> Page 233
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(3) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(18) Page 234
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) 2019264591
<223> phosphorothioate linkage
<400> 290 uuccgcagua uggaucggcu u 21
<210> 291 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) Page 235
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
Page 236
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 291 gccgauccau acugcggaau u 21
<210> 292 2019264591
<211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
Page 237
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (12)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 292 uuccgcagua uggaucggcu u 21
<210> 293 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
Page 238
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(6) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy
<220> <221> misc_feature Page 239
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 293 gccgauccau acugcggaau u 21
<210> 294 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature Page 240
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(9) <223> 2'-deoxy
<220> <221> misc_feature 2019264591
<222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 294 gccgauccau acugcggaau u 21
Page 241
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 295 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature 2019264591
<222> (1)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(18) <223> 2'-O-methyl Page 242
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 295 uuccgcagua uggaucggcu u 21
<210> 296 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl Page 243
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 296 gugguggacu ucucucaauu u 21
<210> 297 <211> 21 Page 244
SIRIFD00001WOPCTSEQ 13 Nov 2019
<212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> Page 245
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (17)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 297 auugagagaa guccaccacu u 21
<210> 298 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> Page 246
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (8)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 298 auugagagaa guccaccacu u 21
<210> 299 <211> 21 <212> RNA <213> Artificial Sequence Page 247
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> 2019264591
<221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) Page 248
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(19) 2019264591
<223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 299 gugguggacu ucucucaauu u 21
<210> 300 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) Page 249
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(4) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
Page 250
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 300 gugguggacu ucucucaauu u 21
<210> 301 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
Page 251
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (4)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(9) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 301 auugagagaa guccaccacu u 21
<210> 302 <211> 21 <212> RNA Page 252
SIRIFD00001WOPCTSEQ 13 Nov 2019
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap 2019264591
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-O-methyl
<220> <221> misc_feature Page 253
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(17) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (18)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 302 ucguggugga cuucucucau u 21
<210> 303 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 254
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (5)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (9)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 303 ugagagaagu ccaccacgau u 21
<210> 304 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature Page 255
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl Page 256
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 304 ugagagaagu ccaccacgau u 21
<210> 305 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(8) <223> 2'-deoxy-2'-fluoro Page 257
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(13) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (14)..(14) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 305 ucguggugga cuucucucau u 21
<210> 306 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic Page 258
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(4) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 306 Page 259
SIRIFD00001WOPCTSEQ 13 Nov 2019
ugagagaagu ccaccacgau u 21
<210> 307 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(16) <223> 2'-deoxy-2'-fluoro
<220> Page 260
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (18)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 307 ucguggugga cuucucucau u 21
<210> 308 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> Page 261
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (4)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(8) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (10)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 308 ccgauccaua cugcggaacu u 21
<210> 309 <211> 21 <212> RNA <213> Artificial Sequence Page 262
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (2)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) Page 263
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 309 guuccgcagu auggaucggu u 21 2019264591
<210> 310 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(9) Page 264
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) 2019264591
<223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
Page 265
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 310 ccgauccaua cugcggaacu u 21
<210> 311 <211> 21 <212> RNA <213> Artificial Sequence
<220> 2019264591
<223> Synthetic
<220> <221> misc_feature <222> (1)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
Page 266
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 311 guuccgcagu auggaucggu u 21
<210> 312 2019264591
<211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
Page 267
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (14)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 312 ccgauccaua cugcggaacu u 21
<210> 313 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(3) <223> 2'-O-methyl
Page 268
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature Page 269
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (20)..(21) <223> phosphorothioate linkage
<400> 313 guuccgcagu auggaucggu u 21
<210> 314 <211> 21 <212> RNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature Page 270
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(21) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (20)..(21) <223> phosphorothioate linkage
<400> 314 guuccgcagu auggaucggu u 21
<210> 315 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(9) <223> 2'-deoxy
<220> <221> misc_feature Page 271
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (10)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> <221> misc_feature 2019264591
<222> (14)..(14) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 315 ccgauccaua cugcggaacu u 21
<210> 316 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature Page 272
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy
<220> <221> misc_feature 2019264591
<222> (3)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage Page 273
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 316 gccgauccau acugcggaau u 21 2019264591
<210> 317 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl Page 274
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 317 uuccgcagua uggaucggcu u 21
<210> 318 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(6) <223> 2'-O-methyl Page 275
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (13)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 318 auugagagaa guccaccacu u 21
<210> 319 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy Page 276
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (3)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (6)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> Page 277
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 319 gugguggacu ucucucaauu u 21
<210> 320 <211> 21 2019264591
<212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(16) <223> 2'-deoxy
<220> Page 278
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (17)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> 2019264591
<221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 320 gugguggacu ucucucaauu u 21
<210> 321 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(10) <223> 2'-deoxy-2'-fluoro
<220> Page 279
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(16) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (17)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 321 auugagagaa guccaccacu u 21
<210> 322 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> Page 280
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (4)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (6)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) Page 281
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(18) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 322 gugguggacu ucucucaauu u 21
<210> 323 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(3) Page 282
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(7) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
Page 283
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 323 auugagagaa guccaccacu u 21
<210> 324 <211> 21 <212> RNA <213> Artificial Sequence
<220> 2019264591
<223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(17) <223> 2'-deoxy-2'-fluoro
Page 284
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage 2019264591
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 324 ccgauccaua cugcggaacu u 21
<210> 325 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
Page 285
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 325 guuccgcagu auggaucggu u 21
<210> 326 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
Page 286
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature Page 287
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (16)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 326 gccgauccau acugcggaau u 21
<210> 327 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 288
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (9)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature 2019264591
<222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 327 uuccgcagua uggaucggcu u 21
<210> 328 <211> 21 <212> RNA <213> Artificial Sequence
Page 289
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature 2019264591
<222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl Page 290
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 328 uuccgcagua uggaucggcu u 21
<210> 329 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy Page 291
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> Page 292
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 329 gccgauccau acugcggaau u 21
<210> 330 <211> 21 2019264591
<212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> Page 293
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 330 uuccgcagua uggaucggcu u 21
<210> 331 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> Page 294
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (5)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (7)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) Page 295
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> inverted abasic cap
<400> 331 gccgauccau acugcggaau u 21
<210> 332 <211> 21 <212> RNA <213> Artificial Sequence 2019264591
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) Page 296
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 332 auugagagaa guccaccacu u 21
<210> 333 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(7) Page 297
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(10) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(14) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 333 gugguggacu ucucucaauu u 21
<210> 334 Page 298
SIRIFD00001WOPCTSEQ 13 Nov 2019
<211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy-2'-fluoro
Page 299
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (15)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 334 ugagagaagu ccaccacgau u 21
<210> 335 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy
Page 300
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (6)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 335 ucguggugga cuucucucau u 21 Page 301
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 336 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(11) <223> 2'-deoxy
<220> <221> misc_feature Page 302
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (15)..(16) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 336 ucguggugga cuucucucau u 21
<210> 337 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(2) <223> 2'-O-methyl
<220> <221> misc_feature Page 303
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (3)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature 2019264591
<222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl Page 304
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 337 ugagagaagu ccaccacgau u 21 2019264591
<210> 338 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy Page 305
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(14) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 338 Page 306
SIRIFD00001WOPCTSEQ 13 Nov 2019
ucguggugga cuucucucau u 21
<210> 339 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> Page 307
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 339 2019264591
ugagagaagu ccaccacgau u 21
<210> 340 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> Page 308
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy
<220> 2019264591
<221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 340 ucguggugga cuucucucau u 21
<210> 341 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> Page 309
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (2)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (10)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 341 ugagagaagu ccaccacgau u 21
<210> 342 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> Page 310
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) Page 311
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 342 guuccgcagu auggaucggu u 21 2019264591
<210> 343 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) Page 312
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) 2019264591
<223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
Page 313
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 343 ccgauccaua cugcggaacu u 21
<210> 344 <211> 21 <212> RNA <213> Artificial Sequence
<220> 2019264591
<223> Synthetic
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(18) <223> 2'-O-methyl
Page 314
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 344 guuccgcagu auggaucggu u 21
<210> 345 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(6) <223> 2'-O-methyl
Page 315
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 345 ccgauccaua cugcggaacu u 21 Page 316
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 346 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 317
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (14)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy
<220> <221> misc_feature 2019264591
<222> (17)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 346 gccgauccau acugcggaau u 21
<210> 347 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature Page 318
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (2)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage Page 319
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 347 uuccgcagua uggaucggcu u 21
<210> 348 <211> 21 <212> RNA <213> Artificial Sequence 2019264591
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl Page 320
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (14)..(14) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 348 gccgauccau acugcggaau u 21
<210> 349 <211> 21 Page 321
SIRIFD00001WOPCTSEQ 13 Nov 2019
<212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy-2'-fluoro
<220> Page 322
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 349 uuccgcagua uggaucggcu u 21
<210> 350 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> Page 323
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(9) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (10)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 350 auugagagaa guccaccacu u 21
<210> 351 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> Page 324
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> 2019264591
<221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) Page 325
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) 2019264591
<223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 351 gugguggacu ucucucaauu u 21
<210> 352 <211> 21 <212> RNA <213> Artificial Sequence
<220> Page 326
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(17) <223> 2'-O-methyl
Page 327
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 352 gugguggacu ucucucaauu u 21
<210> 353 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(8) <223> 2'-O-methyl
Page 328
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (14)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 353 auugagagaa guccaccacu u 21
<210> 354 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(7) <223> 2'-O-methyl
Page 329
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 354 ugagagaagu ccaccacgau u 21
<210> 355 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
Page 330
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature Page 331
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 355 ucguggugga cuucucucau u 21
<210> 356 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy
<220> <221> misc_feature Page 332
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
<220> <221> misc_feature 2019264591
<222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage Page 333
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 356 gccgauccau acugcggaau u 21 2019264591
<210> 357 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl Page 334
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (14)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 357 uuccgcagua uggaucggcu u 21
<210> 358 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy-2'-fluoro Page 335
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> Page 336
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 358 2019264591
gugguggacu ucucucaauu u 21
<210> 359 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> Page 337
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 359 auugagagaa guccaccacu u 21
<210> 360 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(10) <223> 2'-deoxy-2'-fluoro
<220> Page 338
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 360 auugagagaa guccaccacu u 21
<210> 361 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> Page 339
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) Page 340
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(19) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 361 gugguggacu ucucucaauu u 21
<210> 362 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(7) Page 341
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
Page 342
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 362 auugagagaa guccaccacu u 21
<210> 363 <211> 21 <212> RNA <213> Artificial Sequence
<220> 2019264591
<223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
Page 343
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (11)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 363 gugguggacu ucucucaauu u 21
<210> 364 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(5) <223> 2'-deoxy
Page 344
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature Page 345
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (21)..(21) <223> inverted abasic cap
<400> 364 ucguggugga cuucucucau u 21
<210> 365 <211> 21 <212> RNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature Page 346
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(18) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 365 ugagagaagu ccaccacgau u 21
<210> 366 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 347
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (4)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature 2019264591
<222> (8)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl Page 348
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap 2019264591
<400> 366 ucguggugga cuucucucau u 21
<210> 367 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro Page 349
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 367 ugagagaagu ccaccacgau u 21
<210> 368 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(4) <223> 2'-deoxy Page 350
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> Page 351
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> 2019264591
<221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 368 ccgauccaua cugcggaacu u 21
<210> 369 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(10) <223> 2'-O-methyl
<220> Page 352
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (12)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 369 guuccgcagu auggaucggu u 21
<210> 370 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> Page 353
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(8) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (9)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 370 guuccgcagu auggaucggu u 21
<210> 371 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> Page 354
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> 2019264591
<221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) Page 355
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) 2019264591
<223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 371 ccgauccaua cugcggaacu u 21
<210> 372 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) Page 356
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(6) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 372 guuccgcagu auggaucggu u 21
<210> 373 <211> 21 <212> RNA <213> Artificial Sequence
<220> Page 357
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (2)..(2) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy
Page 358
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 373 ccgauccaua cugcggaacu u 21
<210> 374 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
Page 359
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-deoxy
<220> <221> misc_feature Page 360
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature 2019264591
<222> (17)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 374 gccgauccau acugcggaau u 21
<210> 375 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 361
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (3)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(7) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage Page 362
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 375 uuccgcagua uggaucggcu u 21
<210> 376 <211> 21 <212> RNA <213> Artificial Sequence 2019264591
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl Page 363
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (17)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 376 uuccgcagua uggaucggcu u 21
<210> 377 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl Page 364
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 377 Page 365
SIRIFD00001WOPCTSEQ 13 Nov 2019
gccgauccau acugcggaau u 21
<210> 378 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (2)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> Page 366
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (17)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 378 gccgauccau acugcggaau u 21
<210> 379 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(3) <223> 2'-O-methyl
<220> Page 367
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (8)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 379 uuccgcagua uggaucggcu u 21
<210> 380 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> Page 368
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy
<220> 2019264591
<221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) Page 369
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 380 ucguggugga cuucucucau u 21 2019264591
<210> 381 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(14) Page 370
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 381 ugagagaagu ccaccacgau u 21
<210> 382 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) Page 371
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(10) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 382 ugagagaagu ccaccacgau u 21
<210> 383 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) Page 372
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
Page 373
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 383 ucguggugga cuucucucau u 21
<210> 384 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
Page 374
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(11) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 384 guuccgcagu auggaucggu u 21
<210> 385 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
Page 375
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 376
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (17)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 385 ccgauccaua cugcggaacu u 21
<210> 386 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 377
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (10)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature 2019264591
<222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 386 guuccgcagu auggaucggu u 21
<210> 387 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy
<220> <221> misc_feature Page 378
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy
<220> <221> misc_feature 2019264591
<222> (7)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 387 ccgauccaua cugcggaacu u 21
<210> 388 <211> 21 <212> RNA <213> Artificial Sequence
Page 379
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature 2019264591
<222> (2)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl Page 380
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 388 guuccgcagu auggaucggu u 21 2019264591
<210> 389 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro Page 381
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (11)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 389 ccgauccaua cugcggaacu u 21
<210> 390 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic Page 382
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(16) <223> 2'-deoxy-2'-fluoro
<220> Page 383
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy
<220> 2019264591
<221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 390 ccgauccaua cugcggaacu u 21
<210> 391 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> Page 384
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (5)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(8) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (10)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 391 guuccgcagu auggaucggu u 21
<210> 392 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> Page 385
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (2)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) Page 386
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) 2019264591
<223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 392 gccgauccau acugcggaau u 21
<210> 393 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) Page 387
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 393 uuccgcagua uggaucggcu u 21
<210> 394 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) Page 388
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(5) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-deoxy
Page 389
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 394 gccgauccau acugcggaau u 21
<210> 395 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
Page 390
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (10)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(17) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 395 uuccgcagua uggaucggcu u 21
<210> 396 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(10) <223> 2'-O-methyl
Page 391
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (12)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (16)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 396 uuccgcagua uggaucggcu u 21
<210> 397 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy
Page 392
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(6) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature Page 393
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 397 gccgauccau acugcggaau u 21 2019264591
<210> 398 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(10) <223> 2'-O-methyl
<220> <221> misc_feature Page 394
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (11)..(16) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (18)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 398 gccgauccau acugcggaau u 21
<210> 399 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 395
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(8) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 399 uuccgcagua uggaucggcu u 21
<210> 400 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature Page 396
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (2)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy
<220> <221> misc_feature 2019264591
<222> (6)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(18) <223> 2'-O-methyl Page 397
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage 2019264591
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 400 gugguggacu ucucucaauu u 21
<210> 401 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro Page 398
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (16)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 401 auugagagaa guccaccacu u 21
<210> 402 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(7) <223> 2'-deoxy Page 399
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (8)..(8) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> Page 400
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 402 gugguggacu ucucucaauu u 21
<210> 403 <211> 21 2019264591
<212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(13) <223> 2'-deoxy-2'-fluoro
<220> Page 401
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(17) <223> 2'-O-methyl
<220> 2019264591
<221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 403 auugagagaa guccaccacu u 21
<210> 404 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy-2'-fluoro
<220> Page 402
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (10)..(10) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(14) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (16)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 404 auugagagaa guccaccacu u 21
<210> 405 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy
<220> Page 403
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy
<220> 2019264591
<221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(16) Page 404
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) 2019264591
<223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 405 gugguggacu ucucucaauu u 21
<210> 406 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) Page 405
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(6) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
Page 406
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 406 ucguggugga cuucucucau u 21
<210> 407 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
Page 407
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (11)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature Page 408
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (20)..(21) <223> phosphorothioate linkage
<400> 407 ugagagaagu ccaccacgau u 21
<210> 408 <211> 21 <212> RNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy
<220> <221> misc_feature Page 409
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> <221> misc_feature 2019264591
<222> (14)..(14) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 408 ucguggugga cuucucucau u 21
<210> 409 <211> 21 <212> RNA <213> Artificial Sequence
Page 410
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (2)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage Page 411
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 409 ugagagaagu ccaccacgau u 21
<210> 410 <211> 21 <212> RNA <213> Artificial Sequence 2019264591
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro Page 412
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(18) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 410 ugagagaagu ccaccacgau u 21
<210> 411 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(7) <223> 2'-deoxy-2'-fluoro Page 413
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (8)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> Page 414
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 411 ucguggugga cuucucucau u 21
<210> 412 <211> 21 2019264591
<212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(3) <223> 2'-O-methyl
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> Page 415
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (10)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy
<220> 2019264591
<221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 412 ucguggugga cuucucucau u 21
Page 416
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 413 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> 2019264591
<221> misc_feature <222> (1)..(1) <223> 2'-O-methyl
<220> <221> misc_feature <222> (2)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(17) Page 417
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) 2019264591
<223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 413 ugagagaagu ccaccacgau u 21
<210> 414 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (2)..(2) <223> 2'-O-methyl
<220> <221> misc_feature <222> (3)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(8) Page 418
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (9)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) 2019264591
<223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(18) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 414 ugagagaagu ccaccacgau u 21
<210> 415 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) Page 419
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(5) 2019264591
<223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy
Page 420
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (17)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (19)..(21) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 415 ucguggugga cuucucucau u 21
<210> 416 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
Page 421
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (6)..(7) <223> 2'-deoxy
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (9)..(11) <223> 2'-deoxy
<220> <221> misc_feature <222> (12)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 416 cucguggugg acuucucucu u 21
<210> 417 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(8) <223> 2'-O-methyl
Page 422
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (9)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (13)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 417 gagagaaguc caccacgagu u 21
<210> 418 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
Page 423
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature Page 424
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (21)..(21) <223> inverted abasic cap
<400> 418 ccuaugggag ugggccucau u 21
<210> 419 <211> 21 <212> RNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(21) <223> 2'-O-methyl
<220> <221> misc_feature Page 425
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (20)..(21) <223> phosphorothioate linkage
<400> 419 ugaggcccac ucccauaggu u 21
<210> 420 <211> 21 <212> RNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature Page 426
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature 2019264591
<222> (21)..(21) <223> inverted abasic cap
<400> 420 aagaacuccc ucgccucgcu u 21
<210> 421 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature Page 427
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (20)..(21) <223> phosphorothioate linkage
<400> 421 gcgaggcgag ggaguucuuu u 21
<210> 422 <211> 21 <212> RNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(3) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(8) <223> 2'-deoxy
<220> <221> misc_feature <222> (9)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-deoxy
<220> <221> misc_feature Page 428
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> <221> misc_feature 2019264591
<222> (15)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 422 ucugccgauc cauacugcgu u 21
<210> 423 <211> 21 <212> RNA <213> Artificial Sequence
Page 429
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(5) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (6)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-O-methyl
<220> <221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(11) <223> 2'-O-methyl
<220> <221> misc_feature <222> (12)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (17)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage Page 430
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 423 cgcaguaugg aucggcagau u 21
<210> 424 <211> 21 <212> RNA <213> Artificial Sequence 2019264591
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(4) <223> 2'-deoxy
<220> <221> misc_feature <222> (5)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(17) <223> 2'-deoxy Page 431
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 424 agagucuaga cucguggugu u 21
<210> 425 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(6) <223> 2'-deoxy-2'-fluoro Page 432
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (7)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(12) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(15) <223> 2'-O-methyl
<220> <221> misc_feature <222> (16)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 425 caccacgagu cuagacucuu u 21
<210> 426 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap Page 433
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(9) <223> 2'-deoxy
<220> <221> misc_feature <222> (10)..(16) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 426 gaagaagaac ucccucgccu u 21
<210> 427 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(10) <223> 2'-O-methyl Page 434
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (11)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 427 ggcgagggag uucuucuucu u 21
<210> 428 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (3)..(5) <223> 2'-deoxy
<220> <221> misc_feature <222> (6)..(10) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (11)..(13) <223> 2'-deoxy Page 435
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (14)..(15) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (16)..(16) <223> 2'-deoxy 2019264591
<220> <221> misc_feature <222> (17)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(18) <223> 2'-deoxy
<220> <221> misc_feature <222> (19)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 428 ucaagccucc aagcugugcu u 21
<210> 429 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic Page 436
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (4)..(4) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (5)..(6) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (7)..(9) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (10)..(13) <223> 2'-O-methyl
<220> <221> misc_feature <222> (15)..(17) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (18)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 429 gcacagcuug gaggcuugau u 21
<210> 430 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap Page 437
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (1)..(2) <223> 2'-deoxy
<220> <221> misc_feature <222> (3)..(7) <223> 2'-deoxy-2'-fluoro 2019264591
<220> <221> misc_feature <222> (8)..(10) <223> 2'-deoxy
<220> <221> misc_feature <222> (11)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy
<220> <221> misc_feature <222> (14)..(14) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(15) <223> 2'-deoxy
<220> <221> misc_feature <222> (16)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> Page 438
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 430 agccuccaag cugugccuuu u 21
<210> 431 <211> 21 2019264591
<212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (4)..(4) <223> 2'-O-methyl
<220> <221> misc_feature <222> (5)..(5) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (6)..(6) <223> 2'-O-methyl
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (8)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(12) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (13)..(13) <223> 2'-O-methyl
<220> Page 439
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (15)..(17) <223> 2'-O-methyl
<220> <221> misc_feature <222> (18)..(19) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 431 aaggcacagc uuggaggcuu u 21
<210> 432 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (1)..(3) <223> 2'-deoxy
<220> <221> misc_feature <222> (4)..(6) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (7)..(7) <223> 2'-deoxy
<220> Page 440
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (8)..(8) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (9)..(10) <223> 2'-deoxy
<220> 2019264591
<221> misc_feature <222> (11)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(14) <223> 2'-deoxy
<220> <221> misc_feature <222> (15)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 432 agacucgugg uggacuucuu u 21
<210> 433 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> Page 441
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (4)..(5) <223> 2'-O-methyl
<220> <221> misc_feature <222> (6)..(8) <223> 2'-deoxy-2'-fluoro
<220> 2019264591
<221> misc_feature <222> (9)..(9) <223> 2'-O-methyl
<220> <221> misc_feature <222> (10)..(11) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (12)..(12) <223> 2'-O-methyl
<220> <221> misc_feature <222> (13)..(13) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (15)..(16) <223> 2'-O-methyl
<220> <221> misc_feature <222> (17)..(19) <223> 2'-deoxy-2'-fluoro
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 433 agaaguccac cacgagucuu u 21
Page 442
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 434 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> 2019264591
<221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 434 gaagaagaac ucccucgccu u 21
<210> 435 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 435 Page 443
SIRIFD00001WOPCTSEQ 13 Nov 2019
ggcgagggag uucuucuucu u 21
<210> 436 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 436 agacucgugg uggacuucuu u 21
<210> 437 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage Page 444
SIRIFD00001WOPCTSEQ 13 Nov 2019
<400> 437 agaaguccac cacgagucuu u 21
<210> 438 <211> 21 <212> RNA <213> Artificial Sequence 2019264591
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 438 ucugccgauc cauacugcgu u 21
<210> 439 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature Page 445
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (20)..(21) <223> phosphorothioate linkage
<400> 439 cgcaguaugg aucggcagau u 21
<210> 440 <211> 21 <212> RNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 440 aagaacuccc ucgccucgcu u 21
<210> 441 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
Page 446
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 441 gcgaggcgag ggaguucuuu u 21
<210> 442 2019264591
<211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 442 ucaagccucc aagcugugcu u 21
<210> 443 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) Page 447
SIRIFD00001WOPCTSEQ 13 Nov 2019
<223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 443 gcacagcuug gaggcuugau u 21 2019264591
<210> 444 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 444 agccuccaag cugugccuuu u 21
<210> 445 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> Page 448
SIRIFD00001WOPCTSEQ 13 Nov 2019
<221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 445 2019264591
aaggcacagc uuggaggcuu u 21
<210> 446 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 446 cucguggugg acuucucucu u 21
<210> 447 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic Page 449
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage 2019264591
<400> 447 gagagaaguc caccacgagu u 21
<210> 448 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 448 ccuaugggag ugggccucau u 21
<210> 449 <211> 21 <212> RNA <213> Artificial Sequence
Page 450
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature 2019264591
<222> (20)..(21) <223> phosphorothioate linkage
<400> 449 ugaggcccac ucccauaggu u 21
<210> 450 <211> 21 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (1)..(1) <223> inverted abasic cap
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<220> <221> misc_feature <222> (21)..(21) <223> inverted abasic cap
<400> 450 agagucuaga cucguggugu u 21
<210> 451 <211> 21 <212> RNA Page 451
SIRIFD00001WOPCTSEQ 13 Nov 2019
<213> Artificial Sequence
<220> <223> Synthetic
<220> <221> misc_feature <222> (20)..(21) <223> 2'-O-methyl 2019264591
<220> <221> misc_feature <222> (20)..(21) <223> phosphorothioate linkage
<400> 451 caccacgagu cuagacucuu u 21
<210> 452 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 452 ugagagaagu ccaccacga 19
<210> 453 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 453 auugagagaa guccaccac 19
<210> 454 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 454 Page 452
SIRIFD00001WOPCTSEQ 13 Nov 2019
uuccgcagua uggaucggc 19
<210> 455 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<400> 455 guuccgcagu auggaucgg 19
<210> 456 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 456 gaggcauagc agcaggaug 19
<210> 457 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 457 aagaugaggc auagcagca 19
<210> 458 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 458 aaauugagag aaguccacc 19
<210> 459 Page 453
SIRIFD00001WOPCTSEQ 13 Nov 2019
<211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 459 aauugagaga aguccacca 19 2019264591
<210> 460 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 460 gaaguccacc acgagucua 19
<210> 461 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 461 aguauggauc ggcagagga 19
<210> 462 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 462 uccgcaguau ggaucggca 19
<210> 463 <211> 19 <212> RNA <213> Artificial Sequence
Page 454
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<400> 463 aacgccgcag acacaucca 19
<210> 464 <211> 19 <212> RNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<400> 464 aguuccgcag uauggaucg 19
<210> 465 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 465 ccgcguaaag agaggugcg 19
<210> 466 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 466 ccgcaguaug gaucggcag 19
<210> 467 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 467 Page 455
SIRIFD00001WOPCTSEQ 13 Nov 2019
uugagagaag uccaccacg 19
<210> 468 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<400> 468 agaugaggca uagcagcag 19
<210> 469 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 469 gaugaggcau agcagcagg 19
<210> 470 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 470 aaguccacca cgagucuag 19
<210> 471 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 471 augaggcaua gcagcagga 19
<210> 472 Page 456
SIRIFD00001WOPCTSEQ 13 Nov 2019
<211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 472 gagaagucca ccacgaguc 19 2019264591
<210> 473 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 473 ggaguuccgc aguauggau 19
<210> 474 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 474 gcaguaugga ucggcagag 19
<210> 475 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 475 gaguuccgca guauggauc 19
<210> 476 <211> 19 <212> RNA <213> Artificial Sequence
Page 457
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<400> 476 cgaggcgagg gaguucuuc 19
<210> 477 <211> 19 <212> RNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<400> 477 aggcacagcu uggaggcuu 19
<210> 478 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 478 gaggcgaggg aguucuucu 19
<210> 479 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 479 gagugcgaau ccacacucc 19
<210> 480 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 480 Page 458
SIRIFD00001WOPCTSEQ 13 Nov 2019
caguauggau cggcagagg 19
<210> 481 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<400> 481 ggcacagcuu ggaggcuug 19
<210> 482 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 482 aggaguuccg caguaugga 19
<210> 483 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 483 accacgaguc uagacucug 19
<210> 484 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 484 aggcgaggga guucuucuu 19
<210> 485 Page 459
SIRIFD00001WOPCTSEQ 13 Nov 2019
<211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 485 ggagugcgaa uccacacuc 19 2019264591
<210> 486 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 486 aguccaccac gagucuaga 19
<210> 487 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 487 gaagaugagg cauagcagc 19
<210> 488 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 488 uccaccacga gucuagacu 19
<210> 489 <211> 19 <212> RNA <213> Artificial Sequence
Page 460
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<400> 489 guauggaucg gcagaggag 19
<210> 490 <211> 19 <212> RNA 2019264591
<213> Artificial Sequence
<220> <223> Synthetic
<400> 490 gcacuaguaa acugagcca 19
<210> 491 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 491 guccaccacg agucuagac 19
<210> 492 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 492 cacagcuugg aggcuugaa 19
<210> 493 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 493 Page 461
SIRIFD00001WOPCTSEQ 13 Nov 2019
cugaggccca cucccauag 19
<210> 494 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic 2019264591
<400> 494 gagagaaguc caccacgag 19
<210> 495 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 495 ugaggcccac ucccauagg 19
<210> 496 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 496 gcgaggcgag ggaguucuu 19
<210> 497 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 497 cgcaguaugg aucggcaga 19
<210> 498 Page 462
SIRIFD00001WOPCTSEQ 13 Nov 2019
<211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 498 caccacgagu cuagacucu 19 2019264591
<210> 499 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 499 ggcgagggag uucuucuuc 19
<210> 500 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 500 gcacagcuug gaggcuuga 19
<210> 501 <211> 19 <212> RNA <213> Artificial Sequence
<220> <223> Synthetic
<400> 501 aaggcacagc uuggaggcu 19
<210> 502 <211> 19 <212> RNA <213> Artificial Sequence
Page 463
SIRIFD00001WOPCTSEQ 13 Nov 2019
<220> <223> Synthetic
<400> 502 agaaguccac cacgagucu 19
<210> 503 <211> 2270 <212> DNA 2019264591
<213> Hepatitis B virus
<400> 503 gagaacatca catcaggatc cctaggaccc ctgctcgtgt tacaggcggg gtttttcttg 60
ttgacaagaa tcctcacaat accgcagagt ctcgactcgt ggtggacttc tctcaatttt 120
ctagggggat cacccgtgtg tcttggccaa aattcgcagt ccccaacctc caatcactca 180
ccaacctcct gtcctccaat ttgtcctggt tatcgctgga tgtgtctgcg gcgttttatc 240
atattcctct tcatcctgct gctatgcctc atcttcttat tggtacttct ggattatcaa 300
ggtatgttgc ccgtttgtcc tctaattcca ggaaccacaa caaccagtac gggaccctgc 360
aaaacctgca cgactcctgc tcaaggcaac tctatgtttc cctcatgttg ctgtacaaaa 420
cctacggatg gaaattgcac ctgtattccc atcccatcat cttgggcttt cgcaaaatac 480
ctatgggaat gggcctcagt ccgtttctca tggctcagtt cactagtgcc atttgttcag 540
tggttcgtag ggctttcccc cactgtttgg ctttcagtta tgtggatgat gtggtattgg 600
gggccaagtc tgtacaacat cttgagtccc tttataccgc tattaccaat tttcttttgt 660
ctttgggtat acatctaaac cctaacaaaa caaagagatg gggttactcc ctaaacttca 720
tgggatatgt aattggaagt tggggaacat taccacagga tcatattgta caaaaactta 780
aacactgttt tagaaaactt cctgttaata ggcctattga ttggaaagta tgtcaacgaa 840
ttgtgggtct tttgggcttt gccgctcctt ttacacaatg tggataccct gccttaatgc 900
ctttgtatgc atgtatacaa gctaaacagg cttttacttt ctcgccaact tacaaggcct 960
ttctaagtaa acagtatatg aacctttacc ccgttgcccg gcaacggcct ggtctgtgcc 1020
aagtgtttgc tgatgcaacc cccactggct ggggcttggc cattggccat cagcgcatgc 1080
gcggaacctt tgcggctcct ctgccgatcc atactgcgga actcctagca gcttgttttg 1140
ctcgcagcag gtctggagca aaactcatcg ggactgataa ttctgtcgtc ctttctcgga 1200 Page 464
SIRIFD00001WOPCTSEQ 13 Nov 2019
aatatacatc ttttccatgg ctgctaggtt gtactgccaa ctggattctt cgcgggacgt 1260
cctttgttta cgtcccgtcg gcgctgaatc ccgcggacga cccctctcgg ggtcgcttgg 1320
gactctatcg tccccttctc cgtctgccgt tccagccgac cacggggcgc acctctcttt 1380
acgcggtctc cccgtctgtg ccttctcatc tgccggaccg tgtgcacttc gcttcacctc 1440
tgcacgttgc atggagacca ccgtgaacgc ccatcagagc ctgcccaagg tcttacataa 1500 2019264591
gaggactctt ggactctcat caatgtcaac gaccgacctt gaagcttact tcaaagactg 1560
tgtgtttaaa gactgggaag agtcggggga ggaaattagg ttaaaggttt atttattagg 1620
aggctgtagg cataaattgg tctgcgcacc agcaccatgc aactttttca cctctgccta 1680
atcatctctt gtacatgtcc cactgttcaa gcctccaagc tgtgccttgg gtggctttgg 1740
ggcatggaca ttgaccccta taaagaattt ggagctactg tggagttact ctcatttttg 1800
ccttctgact tctttccttc cgtccgggat ctccttgata ccgcctcagc tctgtatcgg 1860
gaagccttag agtctcctga gcattgctca cctcaccata cagcactcag gcaagccatt 1920
ctctgctggg gggaattaat gactctagct acctgggtgg gtaataattt gcaagatcca 1980
gcatccaggg atctagtagt caattatgtt aatactaaca tgggcctaaa gatcaggcaa 2040
ctattgtggt ttcatatttc ttgtcttact tttggaagag atactgtgct tgagtatttg 2100
gtttctttcg gagtgtggat tcgcactcct cctgcctata gaccaccaaa tgcccctatc 2160
ttatcaacac ttccggaaac tactgttgtt agacgaaggg accgaggtag gtcccctaga 2220
agaagaactc cctcgcctcg cagacgaaga tctcaatcgc cgcgtcgcag 2270
<210> 504 <211> 3221 <212> RNA <213> Hepatitis B virus
<400> 504 caugcaacuu uuucaccucu gccuaaucau cucuuguaca ugucccacug uucaagccuc 60
caagcugugc cuuggguggc uuuggggcau ggacauugac ccuuauaaag aauuuggagc 120
uacuguggag uuacucucgu uuuugccuuc ugacuucuuu ccuuccguca gagaucuccu 180
agacaccgcc ucagcucugu aucgagaagc cuuagagucu ccugagcauu gcucaccuca 240
Page 465
SIRIFD00001WOPCTSEQ 13 Nov 2019
ccauacugca cucaggcaag ccauucucug cuggggggaa uugaugacuc uagcuaccug 300
gguggguaau aauuuggaag auccagcauc uagggaucuu guaguaaauu auguuaauac 360
uaacgugggu uuaaagauca ggcaacuauu gugguuucau auaucuugcc uuacuuuugg 420
aagagagacu guacuugaau auuuggucuc uuucggagug uggauucgca cuccuccagc 480
cuauagacca ccaaaugccc cuaucuuauc aacacuuccg gaaacuacug uuguuagacg 540 2019264591
acgggaccga ggcagguccc cuagaagaag aacucccucg ccucgcagac gcagaucucc 600
aucgccgcgu cgcagaagau cucaaucucg ggaaucucaa uguuaguauu ccuuggacuc 660
auaagguggg aaacuuuacg gggcuuuauu ccucuacagu accuaucuuu aauccugaau 720
ggcaaacucc uuccuuuccu aagauucauu uacaagagga cauuauuaau aggugucaac 780
aauuuguggg cccucucacu guaaaugaaa agagaagauu gaaauuaauu augccugcua 840
gauucuaucc uacccacacu aaauauuugc ccuuagacaa aggaauuaaa ccuuauuauc 900
cagaucaggu aguuaaucau uacuuccaaa ccagacauua uuuacauacu cuuuggaagg 960
cugguauucu auauaagcgg gaaaccacac guagcgcauc auuuugcggg ucaccauauu 1020
cuugggaaca agagcuacag caugggaggu uggucaucaa aaccucgcaa aggcaugggg 1080
acgaaucuuu cuguucccaa uccucuggga uucuuucccg aucaucaguu ggacccugca 1140
uucggagcca acucaaacaa uccagauugg gacuucaacc ccgucaagga cgacuggcca 1200
gcagccaacc aaguaggagu gggagcauuc gggccaaggc ucaccccucc acacggcggu 1260
auuuuggggu ggagcccuca ggcucagggc auauugacca cagugucaac aauuccuccu 1320
ccugccucca ccaaucggca gucaggaagg cagccuacuc ccaucucucc accucuaaga 1380
gacagucauc cucaggccau gcaguggaau uccacugccu uccaccaaac ucugcaggau 1440
cccagaguca ggggucugua ucuuccugcu gguggcucca guucaggaac aguaaacccu 1500
gcuccgaaua uugccucuca caucucguca aucuccgcga ggacugggga cccugugacg 1560
aacauggaga acaucacauc aggauuccua ggaccccugc ucguguuaca ggcgggguuu 1620
uucuuguuga caagaauccu cacaauaccg cagagucuag acucguggug gacuucucuc 1680
aauuuucuag ggggaucucc cgugugucuu ggccaaaauu cgcagucccc aaccuccaau 1740
cacucaccaa ccuccugucc uccaauuugu ccugguuauc gcuggaugug ucugcggcgu 1800
Page 466
SIRIFD00001WOPCTSEQ 13 Nov 2019
uuuaucauau uccucuucau ccugcugcua ugccucaucu ucuuauuggu ucuucuggau 1860
uaucaaggua uguugcccgu uuguccucua auuccaggau caacaacaac caguacggga 1920
ccaugcaaaa ccugcacgac uccugcucaa ggcaacucua uguuucccuc auguugcugu 1980
acaaaaccua cggauggaaa uugcaccugu auucccaucc caucguccug ggcuuucgca 2040
aaauaccuau gggagugggc cucaguccgu uucucuuggc ucaguuuacu agugccauuu 2100 2019264591
guucaguggu ucguagggcu uucccccacu guuuggcuuu cagcuauaug gaugaugugg 2160
uauugggggc caagucugua cagcaucgug agucccuuua uaccgcuguu accaauuuuc 2220
uuuugucucu ggguauacau uuaaacccua acaaaacaaa aagauggggu uauucccuaa 2280
acuucauggg cuacauaauu ggaaguuggg gaacuuugcc acaggaucau auuguacaaa 2340
agaucaaaca cuguuuuaga aaacuuccug uuaacaggcc uauugauugg aaaguauguc 2400
aaagaauugu gggucuuuug ggcuuugcug cuccauuuac acaaugugga uauccugccu 2460
uaaugccuuu guaugcaugu auacaagcua aacaggcuuu cacuuucucg ccaacuuaca 2520
aggccuuucu aaguaaacag uacaugaacc uuuaccccgu ugcucggcaa cggccugguc 2580
ugugccaagu guuugcugac gcaaccccca cuggcugggg cuuggccaua ggccaucagc 2640
gcaugcgugg aaccuuugug gcuccucugc cgauccauac ugcggaacuc cuagccgcuu 2700
guuuugcucg cagccggucu ggagcaaagc ucaucggaac ugacaauucu gucguccucu 2760
cgcggaaaua uacaucguuu ccauggcugc uaggcuguac ugccaacugg auccuucgcg 2820
ggacguccuu uguuuacguc ccgucggcgc ugaaucccgc ggacgacccc ucucggggcc 2880
gcuugggacu cucucguccc cuucuccguc ugccguucca gccgaccacg gggcgcaccu 2940
cucuuuacgc ggucuccccg ucugugccuu cucaucugcc gguccgugug cacuucgcuu 3000
caccucugca cguugcaugg agaccaccgu gaacgcccau cagauccugc ccaaggucuu 3060
acauaagagg acucuuggac ucccagcaau gucaacgacc gaccuugagg ccuacuucaa 3120
agacugugug uuuaaggacu gggaggagcu gggggaggag auuagguuaa aggucuuugu 3180
auuaggaggc uguaggcaca aauuggucug cgcaccagca c 3221
<210> 505 <211> 3215 <212> RNA Page 467
SIRIFD00001WOPCTSEQ 13 Nov 2019
<213> Hepatitis B virus
<400> 505 cucaacccag uuccaccaag cucuguugga ucccagggua agggcucugu acuucccugc 60
ugguggcucc aguucaggga cacagaaccc ugcuccgacu auugccucuc ucacaucauc 120
aaucuucucg aagacugggg gcccugcuau gaacauggac aacauuacau caggacuccu 180
aggaccccug cucguguuac aggcggugug uuucuuguug acaaaaaucc ucacaauacc 240 2019264591
acagagucua gacucguggu ggacuucucu caauuuucua gggggacuac ccgggugucc 300
uggccaaaau ucgcaguccc caaccuccaa ucacuuacca accuccuguc cuccaacuug 360
uccuggcuau cguuggaugu gucugcggcg uuuuaucauc uuccucuuca uccugcugcu 420
augccucauc uucuuguugg uucuucugga cuaucaaggu auguugcccg uuuguccucu 480
acuuccagga uccacgacca ccagcacggg acccugcaaa accugcacaa cucuugcaca 540
aggaaccucu auguuucccu ccuguugcug uuccaaaccc ucggacggaa acugcacuug 600
uauucccauc ccaucauccu gggcuuuagg aaaauaccua ugggaguggg ccucagcccg 660
uuucuccugg cucaguuuac uagugcaauu uguucagugg ugcgucgggc uuucccccac 720
uguuuggcuu uuaguuauau ggaugaucug guauuggggg ccaaaucugu gcagcaucuu 780
gagucccuuu auaccgcugu uaccaauuuu cuguuaucug uggguaucca uuuaaauacc 840
ucuaaaacaa aaagaugggg uuacucccua cauuuuaugg guuaugucau ugguaguugg 900
ggaucauuac cccaagauca cauuguacac aaaaucaagg aaugcuuucg aaaacugccu 960
guaaaucguc caauugauug gaaaguuugu caacgcauag ugggucuuuu gggcuuugcu 1020
gccccuuuca cccaaugcgg uuauccugcu cucaugccuc uguaugccug uauuacugcu 1080
aaacaggcuu uugucuucuc gccaaccuac aaggccuuuc uguguaaaca auacaugaac 1140
cuuuacccgg uugcucggca acggccaggc cugugccaag uguuugcuga cgcaaccccc 1200
acugguuggg gcuuggccau uggccaucag cgcaugcgug gaaccuuugu ggcuccucug 1260
ccgauccaua cugcggaacu ccuagcagcu uguuucgcuc gcagcagguc uggagcgacu 1320
cucaucggca cggacaauuc uguuguccuc ucuaggaagu acaccuccuu uccauggcug 1380
cucggaugug cugcaaacug gauccugcgc gggacguccu uuguuuacgu cccaucggcg 1440
cugaaucccg cggacgaccc cucccggggc cgcuuggggc uguaccgccc ucuucuccgu 1500 Page 468
SIRIFD00001WOPCTSEQ 13 Nov 2019
cugccguucc agccgacgac gggucgcacc ucucuuuacg cggacucccc gucuguuccu 1560
ucucaucugc cggaccgugu gcacuucgcu ucaccucugc acgucgcaug gagaccaccg 1620
ugaacgcccc cuggaguuug ccaacagucu uacauaagcg gacucuugga cuuucaggau 1680
ggucaaugac cuggaucgaa gaauacauca aagacugugu auuuaaggac ugggaggagu 1740
ugggggagga gauuagguua aaggucuuug uauuaggagg cuguaggcau aaauuggucu 1800 2019264591
guucaccagc accaugcaac uuuuucaccu cugccuaauc aucuuuuguu caugucccac 1860
uguucaagcc uccaagcugu gccuugggug gcuuuggggc auggacauug acccuuauaa 1920
agaauuugga gcuucugugg aguuacucuc guuuuugccu ucugauuucu ucccaucggu 1980
ucgggaccua cucgacaccg cuucagcucu uuaccgggau gcuuuagagu caccugaaca 2040
uugcacuccc aaccauacug cucucaggca agcuauuuug uguuggggug aguuaaugac 2100
uuuggcuucc ugggugggca auaauuugga ggacccugca gcuagggauu uaguaguuaa 2160
cuauguuaac acuaacaugg gccuaaaaau uagacaacug uugugguuuc acauuuccug 2220
ccuuacuuuu ggaagagaaa caguucuaga guauuuggug uccuuuggag uguggauucg 2280
cacuccuccu gcuuacagac caccaaaugc cccuauccua uccacacuuc cggaaacuac 2340
uguuguuaga cgacgaggca gguccccuag aagaagaacu cccucgccuc gcagacgaag 2400
aucucaaucg ccgcgucgcc gcagaucuca aucuccagcu ucccaauguu aguauuccuu 2460
ggacucauaa ggugggaaac uuuacggggc uuuacucuuc uacugugccu gcuuuuaauc 2520
cuaacugguc cacuccuucu uuuccugaua uucauuugca ucaagaccug auuucuaaau 2580
gugaacaauu uguaggccca cuuacuaaaa augaauuacg aagauuaaaa uugguuaugc 2640
cagcuagauu uuauccuaag guuaccaaau auuuucccau ggauaaaggc aucaaacccu 2700
auuauccuga gcaugcaguu aaucauuacu uuaaaaccag acauuauuug cauacuuuau 2760
ggaaggcggg aauuuuauau aagagagaau ccacacguag cgccucauuu ugugggucac 2820
cauauuccug ggaacaagag cuacagcaug ggagcaccuc ucucaacgac acgaagaggc 2880
augggacaga aucucucugu gcccaauccu cugggauucu uuccagacca ucagcuggau 2940
ccgcuauuca gagcaaauuc cagcaguccc gacugggacu ucaacacaaa caaggacagu 3000
uggccaaugg caaacaaggu aggaguggga ggcuacgguc caggguucac acccccacac 3060 Page 469
SIRIFD00001WOPCTSEQ 13 Nov 2019
gguggccugc ugggguggag cccucaagca caaggugugu uaacaaccuu gccagcagau 3120
ccgccuccug cuuccaccaa ucggcggucc gggagaaagc caaccccagu cucuccaccu 3180
cuaagagaca cacauccaca ggcaaugcag uggaa 3215
<210> 506 <211> 3182 2019264591
<212> RNA <213> Hepatitis B virus
<400> 506 aacuccacaa ccuuccacca aacucugcaa gaucccagag ugagaggccu guauuucccu 60
gcugguggcu ccaguucagg aacaguaaac ccuguucuga cuacugccuc ucccuuaucg 120
ucaaucuccg cgaggacugg ggacccugca cugaacaugg agaacaucac aucaggauuc 180
cuaggacccc ugcucguguu acaggcgggg uuuuucuugu ugacaagaau ccucacaaua 240
ccgcagaguc uagacucgug guggacuucu cucaauuuuc uagggggaac uaccgugugu 300
cuuggccaaa auucgcaguc cccaaccucc aaucacucac caaccucuug uccuccaacu 360
uguccugguu aucgcuggau gugucugcgg cguuuuauca ucuuccucuu cauccugcug 420
cuaugccuca ucuucuuguu gguucuucug gacuaucaag guauguugcc cguuuguccu 480
cuaauuccag gaucuucaac aaccagcacg ggaccaugca gaaccugcac gacuccugcu 540
caaggaaccu cuauguaucc cuccuguugc uguaccaaac cuucggacgg aaauugcacc 600
uguauuccca ucccaucauc uugggcuuuc ggaaaauucc uaugggagug ggccucagcc 660
cguuucuccu ggcucaguuu acuagugcca uuuguucagu gguucguagg gcuuuccccc 720
acuguuuggc uuucaguuau auggaugaug ugguauuggg ggccaagucu guacagcauc 780
uugagucccu uuuuaccgcu guuaccaauu uucuuuuguc ucuggguaua cauuuaaacc 840
cuaacaaaac aaaaagaugg gguuauuccc uaaacuucau ggguuacaua auuggaaguu 900
ggggaacguu gccacaagau cauauuguac aaaagaucaa agaauguuuu agaaaacuuc 960
cuguuaacag gccuauugau uggaaaguau ggcaacgaau ugugggucuu uugggcuuug 1020
cugcuccauu uacacaaugu ggauauccug ccuuaaugcc uuuguaugcc uguauacaag 1080
cuaaacaggc uuucacuuuc ucgccaacuu acaaggccuu ucuaaguaaa caguacauga 1140
Page 470
SIRIFD00001WOPCTSEQ 13 Nov 2019
accuuuaccc cguugcucgg caacggccug gucugugcca aguguuugcu gacgcaaccc 1200
ccacuggcug gggcuuggca auaggcaauc agcgcaugcg uggaaccauu guggcuccuc 1260
ugccgaucca uacugcggaa cuccuagccg cuuguuuugc ucgcagccgg ucuggggcaa 1320
agcucaucgg aacugacaau ucuguugucc ucucgcggaa auauacaucg uuuccauggc 1380
ugcuagguug uacugccaac uggauccuuc gcgggacguc cuuuguuuac gucccgucgg 1440 2019264591
cgcugaaucc cgcggacgac ccuucucggg gccgcuuggg acucucucgu ccccuucucc 1500
gucugccguu ccagccgacc acggggcgca ccucucuuua cgcggucucc ccgucugugc 1560
cuucucaucu gccgguccgu gugcacuucg cuucaccucu gcacauugca uggagaccac 1620
cgugaacgcc caucagauua ugcccaaggu uuuacauaag aggacucuug gacucccagc 1680
aaugucaacg accgaccuug aggccuacuu caaagacugu guguuuaagg acugggagga 1740
gcugggggag gagauuaggu uaaaggucuu uguauuagga ggcuguaggc auaaauuggu 1800
cugcgcacca gcaccaugca acuuuuucac cucugccuaa ucaucucuug uucauguccu 1860
acuguucaag ccuccaagcu gugccuuggg uggcuuuagg gcauggacau ugacccuuau 1920
aaacaauuug gagcuacugu ggaguuacuc ccguauuugc cuucugacuu cuuucucuac 1980
guacgagauc uccuagauac cgccucagcu cuguaucggg aagccuuaga gucuccugag 2040
cauuguucac cucaccauac ugcacucagg caagcaauuc uuugcugggg ggaacuaaug 2100
acucuagcua ccuggguggg uguuaauuug gaagauccag caucuagaga cuuaguaguc 2160
aguuauguca acacuaauau gggccuaaag uucaggcaac ucuugugguu ucacauuucu 2220
ugucucacuu uuggaagaga aacaguuaua gaguauuugg ugucuuucgg aguguggauu 2280
cgcacuccuc cagcuuauag accaccaaau gccccuaucc uaucaacacu uccggagacu 2340
acuguuguua cacgacgagg cagguccccu agaagaagaa cucccucgcc ugccagacca 2400
aggucucaau cgccgcgucg cagaagaucu caaucucggg aaucucaaug uuaguauucc 2460
uuggacucau aaggugggga acuuuacggg gcuuuauucu ucuacuguuc cugucuuuaa 2520
uccucauugg aaaacaccuu cuuuuccuaa uauacauuua caccaagaca uuaucaaaaa 2580
augugaacaa uuuguaggcc cacucacagu caaugagaaa agaagacugc aauugauuau 2640
gccugcuagg uuuuauccaa augucaccaa auauuugcca uuggauaagg guauuaaacc 2700
Page 471
SIRIFD00001WOPCTSEQ 13 Nov 2019
uuauuaucca gagcaucuag uuaaucauua cuuccaaacc agacauuauu uacacacucu 2760
auggaaggcg gguauauuau auaagagaga aacaacacau agcgccucau uuuguggguc 2820
accauauucu ugggaacaag agcuacagca uggggcagaa ucuuuccacc agcaauccuc 2880
ugggauucuu ucccgaccac caguuggauc cagccuucag agcaaacacc gcaaauccag 2940
auugggacuu caaucccaac aaggacaccu ggccagacgc caacaaggua ggagcuggag 3000 2019264591
cauucgggcu ggguuucacc ccaccgcacg gaggucuuuu gggguggagc ccucaggcuc 3060
agggcauacu acauaccgug ccagcaaauc cgccuccugc cucuaccaau cgccagucag 3120
gaaggcagcc uaccccucug ucuccaccuu ugagaaacac ucauccucag gccaugcagu 3180
gg 3182
<210> 507 <211> 3215 <212> RNA <213> Hepatitis B virus
<400> 507 cuccacaaca uuccaccaag cucugcuaga ucccagagug aggggccuau auuuuccugc 60
ugguggcucc aguuccggaa caguaaaccc uguuccgacu acugccucac ccauaucguc 120
aaucuucucg aggacugggg acccugcacc gaacauggag agcacaacau caggauuccu 180
aggaccccug cucguguuac aggcgggguu uuucuuguug acaagaaucc ucacaauacc 240
acagagucua gacucguggu ggacuucucu caauuuucua gggggagcac ccacgugucc 300
uggccaaaau ucgcaguccc caaccuccaa ucacucacca accucuuguc cuccaacuug 360
uccuggcuau cgcuggaugu gucugcggcg uuuuaucaua uuccucuuca uccugcugcu 420
augccucauc uucuuguugg uucuucugga cuaccaaggu auguugcccg uuuguccucu 480
acuuccagga acaucaacua ccagcacggg accaugcaga accugcacga uuccugcuca 540
aggaaccucu auguuucccu cuuguugcug uacaaaaccu ucggacggaa acugcacuug 600
uauucccauc ccaucauccu gggcuuucgc aagauuccua ugggaguggg ccucaguccg 660
uuucuccugg cucaguuuac uagugccauu uguucagugg uucguagggc uuucccccac 720
uguuuggcuu ucagcuauau ggaugaugug guauuggggg ccaagucugu acaacaucuu 780
gagucccuuu uuaccucuau uaccaauuuu cuuuugucuu uggguauaca uuugaacccu 840 Page 472
SIRIFD00001WOPCTSEQ 13 Nov 2019
aauaaaacca aacguugggg cuacucccuu aacuucaugg gauauguaau uggaaguugg 900
gguacuuuac cgcaggaaca uauuguacaa aaacucaagc aauguuuucg aaaauugccu 960
guaaauagac cuauugauug gaaaguaugu caaagaauug ugggucuuuu gggcuuugcu 1020
gccccuuuua cacaaugugg cuauccugcc uugaugccuu uauaugcaug uauacaaucu 1080
aagcaggcuu ucacuuucuc gccaacuuac aaggccuuuc uguguaaaca auaucuaaac 1140 2019264591
cuuuaccccg uugcccggca acggucaggu cucugccaag uguuugcuga cgcaaccccc 1200
acggguuggg gcuuggccau aggccaucgg cgcaugcgug gaaccuuugu ggcuccucug 1260
ccgauccaua cugcggaacu ccuagcagcu uguuuugcuc gcagccgguc uggagcgaaa 1320
cuuaucggaa ccgacaacuc aguuguccuc ucucggaaau acaccuccuu uccauggcug 1380
cuaggcugug cugccaacug gauccugcgc gggacguccu uugucuacgu cccgucggcg 1440
cugaaucccg cggacgaccc gucucggggc cguuugggcc ucuaccgucc ccuucuucau 1500
cugccguucc ggccgaccac ggggcgcacc ucucuuuacg cggucucccc gucugugccu 1560
ucucaucugc cggaccgugu gcacuucgcu ucaccucugc acguagcaug gagaccaccg 1620
ugaacgccca ccaggucuug cccaaggucu uacacaagag gacucuugga cucucagcaa 1680
ugucaacgac cgaccuugag gcauacuuca aagacuguuu guuuaaagac ugggaggagu 1740
ugggggagga gauuagguua aaggucuuug uacuaggagg cuguaggcau aaauuggucu 1800
guucaccagc accaugcaac uuuuuccccu cugccuaauc aucucauguu cauguccuac 1860
uguucaagcc uccaagcugu gccuugggug gcuuuggggc auggacauug acccguauaa 1920
agaauuugga gcuucugugg aguuacucuc uuuuuugccu ucugacuucu uuccuucuau 1980
ucgagaucuc cucgacaccg ccucugcucu guaucgggag gccuuagagu cuccggaaca 2040
uuguucaccu caccauacag cacucaggca agcuauucug uguuggggug aguugaugaa 2100
ucuggccacc ugggugggaa guaauuugga agacccagca uccagggaau uaguagucag 2160
cuaugucaau guuaauaugg gccuaaaaau uagacaacua uugugguuuc acauuuccug 2220
ccuuacuuuu ggaagagaaa cuguccuuga guauuuggug ucuuuuggag uguggauucg 2280
cacuccuccc gcuuacagac caccaaaugc cccuaucuua ucaacacuuc cggaaacuac 2340
uguuguuaga cgacgaggca gguccccuag aagaagaacu cccucgccuc gcagacgaag 2400 Page 473
SIRIFD00001WOPCTSEQ 13 Nov 2019
gucucaaucg ccgcgucgca gaagaucuca aucucgggaa ucucaauguu aguaucccuu 2460
ggacucauaa ggugggaaac uuuacugggc uuuauucuuc uacuguaccu gucuuuaauc 2520
cugauuggaa aacucccucc uuuccucaca uucauuuaca ggaggacauu auuaauagau 2580
gucaacaaua ugugggcccu cugacaguua augaaaaaag gagauuaaaa uuaauuaugc 2640
cugcuagguu cuauccuaac cuuaccaaau auuugcccuu ggacaaaggc auuaaaccgu 2700 2019264591
auuauccuga auaugcaguu aaucauuacu ucaaaacuag gcauuauuua cauacucugu 2760
ggaaggcugg cauucuauau aagagagaaa cuacacgcag cgccucauuu ugugggucac 2820
cauauucuug ggaacaagag cuacagcaug ggagguuggu cuuccaaacc ucgacaaggc 2880
auggggacga aucuuucugu ucccaauccu cugggauucu uucccgauca ccaguuggac 2940
ccugcguucg gagccaacuc aaacaaucca gauugggacu ucaaccccaa caaggaucac 3000
uggccagagg caaaucaggu aggagcggga gcauuugguc caggguucac cccaccacac 3060
ggaggccuuu ugggguggag cccucaggcu cagggcauau ugacaacacu gccagcagca 3120
ccuccuccug ccuccaccaa ucggcaguca ggaagacagc cuacucccau cucuccaccu 3180
cuaagagaca gucauccuca ggccaugcag uggaa 3215
<210> 508 <211> 3215 <212> RNA <213> Hepatitis B virus subtype adw
<400> 508 cuccaccacu uuccaccaaa cucuucaaga ucccagaguc agggcucugu acuuuccugc 60
ugguggcucc aguucaggaa caguaagccc ugcucagaau acugucucag ccauaucguc 120
aaucuuaucg aagacugggg acccugugcc gaacauggag aacaucgcau caggacuccu 180
aggaccccug cucguguuac aggcgggguu uuucuuguug acaaaaaucc ucacaauacc 240
acagagucua gacucguggu ggacuucucu caauuuucua gggggaacac ccgugugucu 300
uggccaaaau ucgcaguccc aaaucuccag ucacucacca accuguuguc cuccaauuug 360
uccugguuau cgcuggaugu gucugcggcg uuuuaucauc uuccucugca uccugcugcu 420
augccucauc uucuuguugg uucuucugga cuaucaaggu auguugcccg uuuguccucu 480
Page 474
SIRIFD00001WOPCTSEQ 13 Nov 2019
aauuccagga ucaucaacca ccagcacggg accaugcaag accugcacaa cuccugcuca 540
aggaaccucu auguuucccu cauguugcug uacaaaaccu auggauggaa acugcaccug 600
uauucccauc ccaucaucuu gggcuuucgc aaaauaccua ugggaguggg ccucaguccg 660
uuucucuugg cucaguuuac uagugccauu uguucagugg uucguagggc uuucccccac 720
ugucuggcuu ucaguuauau ggaugaugug guauuggggg ccaagucugu acaacaucuu 780 2019264591
gagucccuuu augccgcugu uaccaauuuu cuuuugucuu uggguauaca uuuaaacccu 840
aacaaaacaa aaagaugggg auauucccuu aacuucaugg gauauguaau ugggaguugg 900
ggcacauggc cacaggauca uauuguacaa aacuucaaac uauguuuuag aaaacuuccu 960
guaaacaggc cuauugauug gaaaguuugu caacgaauug ugggucuuuu gggguuugcu 1020
gccccuuuua cgcaaugugg auauccugcu uuaaugccuu uauaugcaug uauacaagca 1080
aaacaggcuu uuacuuucuc gccaacuuac aaggccuuuc ucaguaaaca guauaugacc 1140
cuuuaccccg uugcucggca acggccuggu cugugccaag uguuugcuga cgcaaccccc 1200
acugguuggg gcuuggccau aggccaucag cgcaugcgug gaaccuuugu gucuccucug 1260
ccgauccaua cugcggaacu ccuagccgcu uguuuugcuc gcagcagguc uggagcaaac 1320
cucaucggga ccgacaauuc ugucguacuc ucccgcaagu auacaucguu uccauggcug 1380
cuaggcugug cugccaacug gauccugcgc gggacguccu uuguuuacgu cccgucggcg 1440
cugaaucccg cggacgaccc cucccggggc cgcuuggggc ucuaccgccc gcuucuccgu 1500
cugccguacc guccgaccac ggggcgcacc ucucuuuacg cggacucccc gucugugccu 1560
ucucaucugc cggaccgugu gcacuucgcu ucaccucugc acgucgcaug gagaccaccg 1620
ugaacgccca ccggaaccug cccaaggucu ugcauaagag gacucuugga cuuucagcaa 1680
ugucaacgac cgaccuugag gcauacuuca aagacugugu guuuaaugag ugggaggagc 1740
ugggggagga gauuagguua aaggucuuug uacucggagg cuguaggcau aaauuggucu 1800
guucaccagc accaugcaac uuuuucaccu cugccuaguc aucucuuguu cauguccuac 1860
uguucaagcc uccaagcugu gccuugggug gcuuuggggc auggacauug accccuauaa 1920
agaauuugga gcuacugugg aguuacucuc uuuuuugccu ucugacuucu uuccgucggu 1980
gcgggaccuc cuagauaccg ucucugcucu guaucgggaa gccuuaaaau cuccugagca 2040
Page 475
SIRIFD00001WOPCTSEQ 13 Nov 2019
uugcucaccu caccacacag cacucaggca agcuauucug ugcugggggg aauuaaugac 2100
ucuagcuacc ugggugggua auaauuugga agauccagca ucccgggauc uaguagucaa 2160
uuauguuaac acuaacaugg gccuaaagau caggcaacua uggugguuuc acauuuccug 2220
ucuuacuuuu ggaagagaaa cuguucugga auauuuggua ucuuuuggag uguggauucg 2280
cacuccuccu gccuacagac caccaaaugc cccuaucuua ucaacacuuc cggaaacuac 2340 2019264591
uguuguuaga cgacgaggca gguccccuag aagaagaacu cccucgccuc gcagacgaag 2400
gucucaauca ccgcgucgca gaagaucuca aucucgggaa ucccaauguu aguaucccuu 2460
ggacucauaa ggugggaaac uuuacggggc uuuauucuuc uacaguaccu gucuuuaauc 2520
cugaauggca aacuccuucu uuuccagaca uucauuuaca ggaggacauu guugauagau 2580
guaagcaauu ugugggaccc cuuacaguaa augaaaacag gagacuaaaa uuaauaaugc 2640
cugcuagauu uuaucccaau guuaccaaau auuugcccuu agauaaaggu aucaaaccuu 2700
auuauccaga gcauguaguu aaucauuacu uccagacuag acauuauuug cauacucuuu 2760
ggaaggcggg uaucuuauau aaaagagagu caacacauag cgccucauuu ugcgggucac 2820
cuuauucuug ggaacaagau cuacagcaug ggagguuggu cuuccaaacc ucgaaaaggc 2880
auggggacaa aucuuucugu ccccaauccc cugggauucu uccccgauca ucaguuggac 2940
ccugcauuca aagccaacuc agaaaaucca gauugggacc ucaacccaca caaggacaac 3000
uggccggacg cccacaaggu gggaguggga gcauucgggc caggauucac cccuccccau 3060
gggggacugu ugggguggag cccucaggcu cagggcauac ucacaucugu gccagcagcu 3120
ccuccuccug ccuccaccaa ucggcaguca ggacggcagc cuacuccccu aucuccaccu 3180
cuaagggaca cucauccuca ggccaugcag uggaa 3215
<210> 509 <211> 3182 <212> RNA <213> Hepatitis B virus
<400> 509 aacuccacaa ccuuccacca aacucugcaa gaucccagag ugagaggccu guaucucccu 60
gcugguggcu ccaguucagg aacaguaaac ccuguuccga cuacugucuc ucccauaucg 120
ucaaucuucu cgaggauugg ggacccugcg cugaacaugg agaacaucac aucaggauuc 180 Page 476
SIRIFD00001WOPCTSEQ 13 Nov 2019
cuaggacccc ugcucguguu acaggcgggg uuuuucuugu ugacaaaaau ccucacaaua 240
ccgaagaguc uagacucgug guggacuucu cucaauuuuc uagggggaac caccgugugu 300
cuuggccaaa auucgcaguc cccaaccucc aaucacucac caaccuccug uccuccaacu 360
uguccugguu aucgcuggau gugucugcgg cguuuuauca ucuuccucuu cauccugcug 420
cuaugccuca ucuucuuguu gguucuucug gacuaucaag guauguugcc cguuuguccu 480 2019264591
cugauuccag gaucuucaac caccagcacg ggaccaugca gaaccugcac gacuccugcu 540
caaggaaccu cuauguaucc cuccuguugc uguaccaaac cuucggacgg aaauugcacc 600
uguauuccca ucccaucauc cugggcuuuc ggaaaauucc uaugggagug ggccucagcc 660
cguuucuccu ggcucaguuu acuagugcca uuuguucagu gguucguagg gcuuuccccc 720
acuguuuggc uuuuaguuau auggaugaug ugguauuggg ggccaaaacu guucaccauc 780
uugagucccu uuuuaccgcu guuaccaauu uucuuuuguc uuuggguaua caucuaaacc 840
cuaacaaaac aaaaagaugg gguuacucuu uacauuuuau gggcuauguc auuggauguu 900
augggucuuu gccacaagau cacaucauac agaaaaucaa agaauguuuu agaaaacuuc 960
cuguuaacag gccuauugau uggaaagucu gucaacguau ugugggucuu uugggauuug 1020
cugcuccuuu uacacaaugu gguuauccug cuuuaaugcc cuuguaugca uguauucaau 1080
cuaagcaggc uuucacuuuc ucgccaacuu acaaggccuu ucuguguaaa caauaccuga 1140
accuuuaccc cguugcccgg caacgcccag gucugugcca aguguuugcu gacgcaaccc 1200
ccacuggcug gggcuugguc augggccauc agcgcaugcg uggaaccuuu caggcuccuc 1260
ugccgaucca uacugcggaa cuccuagccg cuuguuuugc ucgcagccgg ucuggagcaa 1320
acauucucgg gacggauaac ucuguuguuc ucucccgcaa auauacgucg uuuccauggc 1380
ugcuaggcug ugcugccaac uggauccugc gcgggacguc cuuuguuuac gucccgucgg 1440
cgcugaaucc cgcggacgac ccuucucggg gccgcuuggg acucucucgu ccccuucucc 1500
gucugccguu ucgaccgacc acggggcgca ccucucuuua cgcggacucc ccgucugugc 1560
cuucucaucu gccggaccgu gugcacuucg cuucaccucu gcacgucgca uggagaccac 1620
cgugaacgcc caccaauucu ugcccaaggu cuuacauaag aggacucuug gacucucugu 1680
aaugucaacg accgaccuug aggcauacuu caaagacugu uuguuuaagg acugggagga 1740 Page 477
SIRIFD00001WOPCTSEQ 13 Nov 2019
gucgggggag gagauuagau uaaugaucuu uguacuagga ggcuguaggc auaaauuggu 1800
cugcgcacca gcaccaugca acuuuuucac cucugccuaa ucaucucuug uucauguccu 1860
acuguucaag ccuccaagcu gugccuuggg uggcuuuagg acauggacau ugauccuuau 1920
aaagaauuug gagcuacugu ggaguuacuc ucguuucugc cuucugacuu cuuuccuuca 1980
guacgagauc uucuagauac cgccucagcu cuauaucggg aagccuuaga aucuccugag 2040 2019264591
cauuguucac cucaccauac ugcacucagg caagcaauuc ucugcugggg ggaucuaaua 2100
acucuaucca ccuggguggg ugguaauuug gaagauccaa caucuaggga ccuaguaguc 2160
aguuauguua acacuaauau gggccuaaag uucaggcaac uauugugguu ucacauuucu 2220
ugucucacuu uuggaagaga aacggucaua gaguauuugg ugucuuuugg aguguggauu 2280
cgcacuccuc cagcuuauag accaccaaau gccccuaucu uaucaacacu uccggagacu 2340
acuguuguua gacgacgagg cagguccccu agaagaagaa cucccucgcc ucgcagacga 2400
aggucucaau cgccacgucg cagaagaacu caaucucggg aaucucaaug uuaguauucc 2460
cuggacucau aaggugggaa acuuuacggg gcuuuauucu ucuacuguuc cugucuuuaa 2520
cccucauugg aaaacacccu cuuuuccuaa uauacauuua caccaagaca uuaucaaaaa 2580
augugaacaa uuuguaggcc cacucacagu caaugagaaa agaagacugc aauugauuau 2640
gccugcuagg uuuuauccaa agguuaccaa auauuugcca uuggauaagg guauuaaacc 2700
uuauuaucca gaacaucuag uuaaucauua cuuccaaacc agacauuauu uacacacucu 2760
auggaaggcg gguguauuau auaagagaga aacuacacau agcgccucau uuuguggguc 2820
accauauucc ugggaacaag agcuacagca uggggcagaa ucuuuccacc agcaauccuc 2880
ugggauucuu ucccgaccac caguuggauc cagccuucag agcaaacacu gcaaauccag 2940
auugggacuu caaucccaac aaggacuccu ggccagacgc caacaaggua ggagcuggag 3000
cauucgggcu gggauucacc ccaccgcacg gaggccuuuu gggguggagc ccucaggcuc 3060
agggcauacu acaaaccuug ccagcaaauc cgccuccugc cuccaccaau cgccagucag 3120
gaaggcaacc uaccccucug ucuccaccuu ugagaaacac ucauccucag gccaugcagu 3180
gg 3182
Page 478
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 510 <211> 3212 <212> RNA <213> Hepatitis B virus
<400> 510 uuccacaaca uuccaccaag cucugcagga ucccagagua agaggccugu auuuuccugc 60
ugguggcucc aguuccggaa cagugaaccc uguuccgacu acugccucac ucaucucguc 120 2019264591
aaucuucucg aggauugggg acccugcacc gaacauggaa agcaucacau caggauuccu 180
aggaccccug cucguguuac aggcgggguu uuucuuguug acaaaaaucc ucacaauacc 240
gcagagucua gacucguggu ggacuucucu caauuuucua gggggagcuc ccgugugucu 300
uggccaaaau ucgcaguccc caaccuccag ucacucacca accucuuguc cuccaauuug 360
uccuggcuau cgcuggaugu gucugcggcg uuuuaucauc uuccucuuca uccugcugcu 420
augccucauc uucuuguugg uucuucugga cuaucaaggu auguugcccg uuuguccucu 480
aauuccagga ucaucaacca ccaguacggg acccugccga accugcacga cucuugcuca 540
aggaaccucu auguuucccu cauguugcug uucaaaaccu ucggacggaa auugcacuug 600
uauucccauc ccaucaucau gggcuuucgg aaaauuccua ugggaguggg ccucagcccg 660
uuucuccugg cucaguuuac uagugccauu uguucagugg uucgccgggc uuucccccac 720
ugucuggcuu ucaguuauau ggaugaugug guauuggggg ccaagucugu acaacaucuu 780
gagucccuuu auaccgcugu uaccaauuuu cuuuugucuu uggguauaca uuuaaauccc 840
aacaaaacaa aaagaugggg cuauucccuu aauuucaugg guuauguaau uggaaguugg 900
ggcucauuac cacaggaaca caucauacaa aaaaucaaag acuguuuuag aaaacucccu 960
guuaaccggc cuauugauug gaaaguaugu caaagaauug ugggucuuuu gggcuuugcu 1020
gcccccuuua cacaaugugg auauccugcu uuaaugccuc uguaugcaug uacucaaucu 1080
aagcaggcuu ucacuuucuc gccaacuuac aaggccuuuc uguguaaaca auaccugaac 1140
cuuuaccccg uugcccggca acggccaggu cugugccaag uguuugcuga ugcaaccccc 1200
acuggcuggg gcuuggccau aggcauucag cgcaugcgcg gaaccuuugu ggcuccucug 1260
ccgauccaua cugcggaacu ccuagccgcu uguuuugcuc gcagcagguc uggagcaaaa 1320
cuuaucggga ccgauaauuc ugucguucuc ucccggaagu auacauccuu uccauggcug 1380
Page 479
SIRIFD00001WOPCTSEQ 13 Nov 2019
cuaggcugug cugccaacug gauccugcga gggacguccu uugucuacgu cccgucagcg 1440
cugaauccug cggacgaccc gucucggggu cgcuugggga ucuaucgucc ccuucuccgu 1500
cugccguucc agccgaccac ggggcgcacc ucucuuuacg cggucucccc gucuguuccu 1560
ucucaucugc cggaccgugu gcacuucgcu ucaccucugc acgucgcaug gagaccaccg 1620
ugaacgccca ccaaauauug cccaaggucu uacauaagag gacucuugga cucucugcaa 1680 2019264591
ugucaacgac cgaccuugag gcauacuuca aagacuguuu guuuaaagac ugggaggagu 1740
cgggggagga gauuagauua aaggucuuug uacuaggagg cuguaggcau aaauuggucu 1800
gcgcaccagc accaugcaac uuuuucaccu cugccuaauc aucucuuguu cauguccuac 1860
uguucaagcc uccaagcugu gccuugggug gcuuuggggc auggacauug acccuuauaa 1920
agaauuugga gcuacugugg aguuacucuc guuuuugccu ucugacuucu uuccuucagu 1980
aagagaucuu cuagauaccg ccucagcucu guaucgggau gccuuagagu cuccugagca 2040
uuguucaccu caccacacug cacucaggca agccauucuu ugcuggggag aacuaaugac 2100
ucuagcuacc ugggugggug uaaauuugga agauccagca uccagggacc uaguagucag 2160
uuaugucaau acuaauaugg gccuaaaguu caggcaauua uugugguuuc acauuucuug 2220
ucucacuuuu ggaagagaaa ccgucauaga guauuuggug ucuuuuggag uguggauucg 2280
cacuccucca gcuuauagac caccaaaugc cccuaucuua ucaacacuuc cggagaauac 2340
uguuguuaga cgaagaggca gguccccuag aagaagaacu cccucgccuc gcagacgaag 2400
aucucaaucg ccgcgucgca gaagaucuca aucuccagcu ucccaauguu aguauuccuu 2460
ggacucauaa ggugggaaau uuuacggggc uuuacucuuc uacuauaccu gucuuuaauc 2520
cuaacuggaa aacuccaucu uuuccugaua uucauuugca ccaggacauu auuaacaaau 2580
gugaacaauu uguagguccu cuaacuguaa augaaaaacg aagauuaaac uuagucaugc 2640
cugcuagauu uuuucccauc ucuacgaaau auuugccccu agagaaaggu auaaaaccuu 2700
auuauccaga uaauguaguu aaucauuacu uccaaaccag acacuauuua cauacccuau 2760
ggaaggcggg caucuuauau aaaagagaaa cuacacguag cgccucauuu ugugggucac 2820
cuuauucuug ggaacaagag cuacaucaug gggcuuucuu ggacgguccc ucucgaaugg 2880
gggaagaaua uuuccaccac caauccucug ggauuuuuuc ccgaccacca guuggaucca 2940
Page 480
SIRIFD00001WOPCTSEQ 13 Nov 2019
gcauucagag caaacaccag aaauccagau ugggaccaca aucccaacaa agaccacugg 3000
acagaagcca acaagguagg agugggagca uucgggccug gguucacucc cccacacgga 3060
ggccuuuugg gguggagccc ucaggcucaa ggcaugcuaa aaacauugcc agcagauccg 3120
ccuccugccu ccaccaaucg gcagucagga aggcagccua ccccaaucac uccaccuuug 3180
agagacacuc auccucaggc caugcagugg aa 3212 2019264591
<210> 511 <211> 3215 <212> RNA <213> Hepatitis B virus
<400> 511 cucgaggacu ggggacccug caccgaacau ggagagcaca acaucaggau uccuaggacc 60
ccugcucgug uuacaggcgg gguuuuucuu guugacaaga auccucacaa uaccacagag 120
ucuagacucg ugguggacuu cucucaauuu ucuaggggga gcacccacgu guccuggcca 180
aaauucgcag uccccaaccu ccaaucacuc accaaccucu uguccuccaa uuuguccugg 240
uuaucgcugg augugucugc ggcguuuuau cauauuccuc uucauccugc ugcuaugccu 300
caucuucuug uugguucuuc uggacuacca agguauguug cccguuuguc cucuacuucc 360
aggaacauca acuaccagca cgggaccaug caagaccugc acgauuccug cucaaggaac 420
cucuauguuu cccucuuguu gcuguacaaa accuucggac ggaaacugca cuuguauucc 480
caucccauca uccugggcuu ucgcaagauu ccuaugggag ugggccucag uccguuucuc 540
cuggcucagu uuacuagugc cauuuguuca gugguucgca gggcuuuccc ccacuguuug 600
gcuuucaguu auauggauga ugugguauug ggggccaagu cuguacaaca ucuugagucc 660
cuuuuuaccu cuauuaccaa uuuucuuuug ucuuugggua uacauuugaa cccuaauaaa 720
accaaacguu ggggcuacuc ccuuaacuuc augggauaug uaauuggaag uugggguacu 780
uuaccacagg aacauauugu auuaaaacuc aagcaauguu uucgaaaacu gccuguaaau 840
agaccuauug auuggaaagu augucaaaga auuguggguc uuuugggcuu ugcugccccu 900
uuuacacaau guggcuaucc ugccuugaug ccuuuguaug cauguauaca aucuaagcag 960
gcuuucacuu ucucgccaac uuauaaggcc uuucuguguc aacaauaccu gcaccuuuac 1020
cccguugccc ggcaacgguc aggucucugc caaguguuug cugacgcaac ccccacugga 1080 Page 481
SIRIFD00001WOPCTSEQ 13 Nov 2019
uggggcuugg ccauaggcca ucggcgcaug cguggaaccu uugugguucc ucugccgauc 1140
cauacugcgg aacuccuagc agcuuguuuu gcucgcgacc ggucuggagc aaaacuuauc 1200
gggacugaca acucgguugu ccucucucgg aaauacaccu ccuucccaug gcugcucggg 1260
ugugcugcca acuggauccu gcgcgggacg uccuuugucu acgucccguc ggcgcugaau 1320
cccgcggacg acccgucucg gggccguuug ggccucuacc gucccuugcu uucucugccg 1380 2019264591
uuccagccga ccacggggcg caccucucuu uacgcggucu ccccgucugu gccuucucau 1440
cugccggacc gugugcacuu cgcuucaccu cugcacgucg cauggagacc accgugaacg 1500
gccaccaggu cuugcccaag cucuuacaua agaggacucu uggacucuca gcaaugucaa 1560
caaccgaccu ugaggcauac uucaaagacu guuuguuuaa agacugggag gaguuggggg 1620
aggagauuag guuaaagguc uuuguacuag gaggcuguag gcauaaauug gucuguucac 1680
cagcaccaug caacuuuuuc accucugccu aaucaucuca uguucauguc cuacuguuca 1740
agccuccaag cugugccuug gguggcuuug gggcauggac auugacccgu auaaagaauu 1800
uggagcuucu guggaguuac ucucuuuuuu gccuucugac uucuuuccuu cuauucgaga 1860
ucuccucgac accgccucug cucuguaucg ggaggccuua gagucuccgg aacauuguuc 1920
accucaccau acagcacuca ggcaagcuau ucuguguugg ggugaguuga ugaauuuggc 1980
caccugggug ggaaguaauu uggaagaccc agcauccagg gaauuaguag ucagcuaugu 2040
caauguuaau augggccuaa aaaucagaca acuauugugg uuucauauuu ccugucuuac 2100
uuuuggaaga gaaacuguuc uugaguauuu ggugucuuuu ggagugugga uucgcacucc 2160
ucccgcuuac agaccaccaa augccccuau cuuaucaaca cuuccggaaa cuacuguugu 2220
uagacgacga ggcagguccc cuagaagaag aacucccucg ccucgcagac gaaggucuca 2280
aucgccgcgu cgcagaagau cucaaucucg ggaaucucaa uguuaguauc ccuuggacuc 2340
auaagguggg aaacuuuacu gggcuuuauu cuucuacugu accuguccuu aauccugagu 2400
cccaaacucc cuccuuuccu aacauucauu uacaggagga cauuauuaau agaugucaac 2460
aauauguggg cccucuuaca guuaaugaaa aaaggagauu aaaauuaauu augccugcua 2520
gguucuaucc uaaccuuacc aaauauuugc ccuuggauaa aggcauuaaa ccuuauuauc 2580
cugaacaugc aguuaaucau uacuucaaaa cuaggcauua uuuacauacu cuguggaagg 2640 Page 482
SIRIFD00001WOPCTSEQ 13 Nov 2019
cuggcauucu auauaaaaga gaaacuacac gcagcgcuuc auuuuguggg ucaccauauu 2700
cuugggaaca agagcuacgg caugggaggu uggucuucca aaccucgaca aggcaugggg 2760
acgaaucuuu cuguucccaa uccucuggga uucuuucccg aucaccaguu ggacccugcg 2820
uucggagcca acucaaacaa uccagauugg gacuucaacc ccaacaagga ucacuggcca 2880
gaggcaauca agguaggagc gggagacuuc gggccagggu ucaccccacc acacggcggu 2940 2019264591
cuuuuggggu ggagcccuca ggcucagggc auauugacaa cagugccagc agcgccuccu 3000
ccuguuucca ccaaucggca gucaggaaga cagccuacuc ccaucucucc accucuaaga 3060
gacagucauc cucaggccau gcaguggaac uccacaacau uccaccaagc ucugcuagau 3120
cccagaguga ggggccuaua uuuuccugcu gguggcucca guuccggaac aguaaacccu 3180
guuccgacua cugucucacc cauaucguca aucuu 3215
<210> 512 <211> 3182 <212> RNA <213> Hepatitis B virus
<400> 512 aacuccacaa ccuuccaaca cacucugcaa gaucccagag ugaaaggccu guaucucccu 60
gcugguggcu ccaguucagg aacaguaaac ccuguuccga ccacugccuc ucccuuaucg 120
ucaaucuucu cgaggauugg ggacccugcg cugaacaugg agaacaucac aucaggauuc 180
cuaggacccc uucucguguu acaggcgggg uuuuucuugu ugacaagaau ccucacaaua 240
ccgcagaguc uagacucgug guggacuucu cucaauuuuc uagggggaac uaccgugugu 300
cuuggccaaa auucgcaguc cccaaccucc aaucacucac caaccuccug uccuccaacu 360
uguccugguu aucgcuggau gugucugcgg cguuuuauca ucuuccucuu cauccugcug 420
cuaugccuca ucuucuuguu gguucuucug gacuaucaag guauguugcc cguuuguccu 480
cuaauuccag gauccucaac caccagcacg ggaccaugcc gcaccugcac gacuccugcu 540
caaggaaccu cuauguaucc cuccuguugc uguaccaaac cuucggacgg aaauugcacc 600
uguauuccca ucccaucauc uugggcuuuc ggaaaauucc uaugggagug ggccucagcc 660
cguuucuccu ggcucaguuu acuagugcca uuuguucagu gguucguagg gcuuuccccc 720
Page 483
SIRIFD00001WOPCTSEQ 13 Nov 2019
acuguuuggc uuucaguuau auggaugaug ugguauuggg ggccaagucu guacagcauc 780
uugagucccu uuuuaccgcu guaaccaauu uucuucuguc uuuggguaua cauuuaaacc 840
cuaacaaaac aaagagaugg gguuacucuc uaaauuuuau gggguauauc auuggauguc 900
auggguccuu gccacaagaa cacaucguac aaaaaaucaa agaauguuuu aggaaacuuc 960
cuauuaacag gccuauugau uggaaagugu gucaacgaau ugugggucuu uuggguuuug 1020 2019264591
cugccccuuu uacacaaugu gguuacccug cuuuaaugcc cuuguaugca uguauccaau 1080
cuaagcaggc uuucacuuuc ucgucaacuu acaaggccuu ucuguguaaa caauaccuga 1140
accuuuaccc cguugcccgg caacggccag gucugugcca aguguuugcu gacgcaaccc 1200
ccacugguug gggcuuggcc augggccauc agcgcaugcg uggaaccuuu cuggcuccuc 1260
ugccgaucca uacugcggaa cuccuagccg cuuguuuugc ucgcagcagg ucuggagcaa 1320
acauuaucgg gacugauaac ucuguugucc ucucccgcaa auauacaucg uuuccauggc 1380
ugcuaggcug ugcugccaac uggauccugc gcgggacguc cuuuguuuac gucccgucgg 1440
cgcugaaucc cgcggacgac cccucucggg gucgcuuggg acuuucucgu ccccuucucc 1500
gucugccguu ccgaccgacc acggggcgca ccucucuuua cgcggacucc ccgucugugc 1560
cuucucaucu gccggaccga gugcacuuag guucaccucu gcacgucgca uggagaccac 1620
cgugaacgcc caccaaaucu ugcccaaggu cuuacauaag aggacucuug gacuuucugc 1680
aaugucaacg accgaccuug aggcauacuu caaagacugu uuguuuaaag acugggagga 1740
guugggggag gagauuuggu uaaaggucuu uguacuagga ggcuguaggc auaaauuggu 1800
cugcgcacca gcaccaugcg acuuuuucac cucugccuaa ucaucucuug uucauguccu 1860
acuguucaag ccucuaagcu gugccuuagg uggcuuuagg gcauggacau ugacccuuau 1920
aaagaauuug gagcuucugu ggaguuacuc ucguuuuugc cuucugacuu cuuuccuuca 1980
guacgagauc uucuagauac cgccucagcu cuguaucggg aagccuuaga gucuccugag 2040
cacuguacac cucaccauac ugcacucagg caagcaauuc uuugcugggg ggaccuaaug 2100
acucuagcua ccuggguggg uguuaauuug gaagauccag caucuagaga ccuaguaguu 2160
aguuauguca acacuaauau gggccuaaag uucaggcaac uuuugugguu ucacguuucu 2220
ugucucacuu uuggaagaga aacaguugua gaguauuugg ugucuuucgg aguguggauu 2280
Page 484
SIRIFD00001WOPCTSEQ 13 Nov 2019
cgcacuccuc cagcuuauag accaccaaau gccccuaucc uaucaacacu uccggagacu 2340
acuguuguua gacgacgaug cagguccccu agaagaagaa cucccucgcc ucgcagacga 2400
aggucucaau cgccgcgucg cagaagaucu caaucucggg aaucucaaug uuaguauucc 2460
uuggacucau aaggugggga acuuuacugg gcuuuauucu ucuacuguac cugucuuuaa 2520
uccucauugg aaaacaccau cuuuuccuaa uauacauuua caccaagaca uuaucaaaaa 2580 2019264591
augugaacag uuuguaggcc cacucacagu uaaugagaaa agaagauugc aauugauuau 2640
gccugcuagg uuuuauccaa auguuaccaa auauuuacca uuggauaagg guauuaaacc 2700
uuauuaucca gaacaucuag uuaaucauua cuuccaaacu agacacuauu uacacacucu 2760
auggaaggcg gguauauuau auaagagaga aacaacacau agcgccucau uuuguggguc 2820
accauauucu ugggaacaag aucuacagca uggggcagaa ucuuuccacc agcaauccuc 2880
ugggauucuu ucccgaccac caguuggauc cagccuucag agcaaacacc gcaaauccag 2940
auugggacuu caaucccaac aaggacaccu ggccagaugc caacaaggua ggagcuggag 3000
cauucgggcu ggguuucacc ccaccgcacg gaggccuuuu gggauggagc ccucaggcuc 3060
agggcauacu acuaacuuug ccagcaaauc cgccuccugc cuccaccaau cgccagucag 3120
gcaggcagcc uaccccgcug acuccaccuu ugagaaacac ucauccucag gccaugcagu 3180
gg 3182
<210> 513 <211> 3221 <212> RNA <213> Hepatitis B virus
<400> 513 uuccacugcc uuccaccaag cucugcaaga ccccagaguc aggggucugu auuuuccugc 60
ugguggcucc aguucaggaa caguaaaccc ugcuccgaau auugccucuc acaucucguc 120
aaucuccgcg aggaccgggg acccugugac gaacauggag aacaucacau caggauuccu 180
aggaccccug cccguguuac aggcgggguu uuucuuguug acaagaaucc ucacaauacc 240
gcagagucua gacucguggu ggacuucucu caauuuucua gggggaucac ccgugugucu 300
uggccaaaau ucgcgauccc caaccuccaa ucacucacca accuccuguc cuccaauuug 360
uccugguuau cgcuggaugu gucugcggcg uuuuaucaua uuccucuuca uccugcugcu 420 Page 485
SIRIFD00001WOPCTSEQ 13 Nov 2019
augccucauc uucuuauugg uucuucugga uuaucaaggu auguugcccg uuuguccucu 480
aauucuagga ucaacaacaa ccaguacggg accaugcaaa accugcacga cuccugcuca 540
aggcaacucu auguuucccu cauguugcug uacaaaaccu acggauggaa auugcaccug 600
uauucccauc ccaucgucuu gggcuuucgc aaaauaccua ugggaguggg ccucaguccg 660
uuucucuugg cucaguuuac uagugccauu uguucagugg uucguagggc uuucccccac 720 2019264591
uguuuggcuu ucagcuauau ggaugaugug guauuggggg ccaagucugu acagcaucgu 780
gaguuccuuu auaccgcugu uaccaauuuu cuuuugucuc uggguauaca uuuaaacccu 840
aacaaaacaa aaagaugggg uuauucccua aacuucaugg guuauguaau uggaaguugg 900
ggaacauugc cacaggauca uauuguacaa aaaaucaaac acuguuuuag aaaacuuccu 960
guuaacaggc cuauugauug gaaaguaugu caaagaauug ugggucuuuu gggcuuugcu 1020
gcuccuuuua cacaaugugg auauccugcc uuaaugcccu uguaugcaug uauacaagcu 1080
aaacaggcuu ucacuuucuc gccaacuuac aaggccuuuc uaaguaaaca guacaugaac 1140
cuuuaccccg uugcucggca acggccuggu cugugccaag uauuugcuga ugcaaccccc 1200
acuggcuggg gcuuggccau aggccaucag cgcaugcgcg gaaccuuugu ggcuccucug 1260
ccgauccaua cugcggaacu ccuagccgcu uguuuugcuc gcagccgguc uggagcgaaa 1320
cucaucggaa cugacaauuc ugucguccuc ucgcggaaau auaccucguu uccauggcua 1380
cuaggcugug cugccaacug gauccuucgc gggacguccu uuguuuacgu cccgucggcg 1440
cugaaucccg cggacgaccc cucucggggc cgcuugggac ucucucgucc ccuucuccgu 1500
cugccguucc agccgaccac ggggcgcacc ucucuuuacg cggucucccc gucugugccu 1560
ucucaucugc cgguccgugu gcacuucgcu ucaccucugc acguugcaug gagaccaccg 1620
ugaacgccca ucagauccug cccaaggucu uacauaagag gacucuugga cucccagcaa 1680
ugucaacgac cgaccuugag gccuacuuca aagacugugu guuuaaggac ugggaggagc 1740
ugggggagga gauuagguua aaggucuuug uauuaggagg cuguaggcau aaauuggucu 1800
gcgcaccagc accaugcaac uuuuucaccu cugccuaauc aucucuugua caugucccac 1860
uguucaagcc uccaagcugu gccuugggug gcuuuggggc auggacauug acccuuauaa 1920
agaauuugga gcuacugugg aguuacucuc guuuuugccu ucugacuucu uuccuuccgu 1980 Page 486
SIRIFD00001WOPCTSEQ 13 Nov 2019
cagagaucuc cuagacaccg ccucggcucu guaucgggaa gccuuagagu cuccugagca 2040
uugcucaccu caccauaccg cacucaggca agccauucuc ugcugggggg aauugaugac 2100
ucuagcuacc ugggugggua auaauuugga agauccagca uccagggauc uaguagucaa 2160
uuauguuaau acuaacaugg gauuaaagau caggcaacuc uugugguuuc auaucucuug 2220
ccuuacuuuu ggaagagaaa cuguacuuga auauuugguc ucuuucggag uguggauucg 2280 2019264591
cacuccucca gccuauagac caccaaaugc cccuaucuua ucaacacuuc cggaaacuac 2340
uguuguuaga cgacgggacc gaggcagguc cccuagaaga agaacucccu cgccucgcag 2400
acgcagaucu caaucgccgc gucgcagaag aucucaaucu cgggaaucuc aauguuagua 2460
uuccuuggac ucauaaggug ggaaacuuca cugggcuuua uuccucuaca gcaccuaucu 2520
uuaauccuga auggcaaacu ccuuccuuuc cuaaaauuca uuuacaagag gacauuauua 2580
auagguguca acaauuugug ggcccucuca cuguaaauga aaagagaaga uugaaauuaa 2640
uuaugccugc uagauucuau ccuacccaca cuaaauauuu gcccuuagac aaaggaauua 2700
aaccuuauua uccagaucag guaguuaauc auuacuucca aaccagacau uauuuacaua 2760
cucuuuggaa ggcggguauu cuauauaaga gagaaaccac acguagcgca ucauuuugcg 2820
ggucaccaua uucuugggaa caagagcuac agcaugggag guuggucauc aaaaccucgc 2880
aaaggcaugg ggacgaaucu uucuguuccc aacccucugg gauucuuucc cgaucaucag 2940
uuggacccug uauucggagc caacucaaac aauccagauu gggacuucaa ccccaucaag 3000
gaccacuggc cagcagccaa ccagguagga gugggagcau ucgggccagg guucaccccu 3060
ccacacggcg guguuuuggg guggagcccu caggcucagg gcauguugac cccaguguca 3120
acaauuccuc cuccugccuc cgccaaucgg cagucaggaa ggcagccuac ucccaucucu 3180
ccaccucuaa gagacaguca uccucaggcc augcagugga a 3221
<210> 514 <211> 3215 <212> RNA <213> Hepatitis B virus
<220> <221> misc_feauure Page 487
SIRIFD00001WOPCTSEQ 13 Nov 2019
<222> (2768)..(2768) <223> n is a, c, g, or u
<400> 514 cuccaccaca uuccaccaag cucugcuaca ccccagagua aggggccuau acuuuccugc 60
ugguggcucc aguuccggaa caguaaaccc uguuccgacu acugccucuc ccauaucguc 120
aaucuucucg aggacugggg acccugcacc gaacauggag aacacaacau caggauuccu 180 2019264591
aggaccccug cucguguuac aggcgggguu uuucuuguug acaagaaucc ucacaauacc 240
acagagucua gacucguggu ggacuucucu caauuuucua gggggagcac ccacgugucc 300
uggccaaaau ucgcaguccc caaccuccaa ucacucacca accucuuguc cuccaauuug 360
uccuggcuau cgcuggaugu gucugcggcg uuuuaucaua uuccucuuca uccugcugcu 420
augccucauc uucuuguugg uucuucugga cuaccaaggu auguugcccg uuuguccucu 480
acuuccagga acaucaacua ccagcacggg accaugcaag accugcacga uuccugcuca 540
aggcaccucu auguuucccu cuuguugcug uacaaaaccu ucggacggaa acugcacuug 600
uauucccauc ccaucauccu gggcuuucgc aagauuccua ugggaguggg ccucaguccg 660
uuucuccugg cucaguuuac uagugccauu uguucagugg uucguagggc uuucccccac 720
uguuuggcuu ucaguuauau ggaugaugug guauuggggg ccaagucugu acaacaucuu 780
gagucccuuu uuaccgcugu uaccaauuuu cuuuugucuu uggguauaca uuugaacccu 840
aauaaaacca aacguugggg uuacucccuu aacuucaugg gauauguaau uggaaguugg 900
gguacuuuac cgcaagacca uauuguacua aaaaucaagc aauguuuucg aaaacugccu 960
guaaauagac cuauugauug gaaaguaugu cagagaauug ugggucuuuu gggcuuugcu 1020
gccccuuuua cacaaugugg cuauccugcc uuaaugccuu uauaugcaug uauacaaucu 1080
aagcaggcuu ucacuuucuc gccaacuuac aaggccuuuc uguguaaaca auaucugaac 1140
cuuuaccccg uugcccggca acggucaggu cucugccaag uguuugcuga cgcaaccccc 1200
acuggauggg gcuuggcuau uggccaucgc cgcaugcgug gaaccuuugu ggcuccucug 1260
ccgauccaua cugcggaacu ccuagcagcu uguuuugcuc gcagccgguc uggagcgaaa 1320
cugaucggaa cggacaacuc uguuguucuc ucucggaaau acaccuccuu uccauggcug 1380
cuagggugug cugccaacug gauccugcgc gggacguccu uuguuuacgu cccgucggcg 1440
Page 488
SIRIFD00001WOPCTSEQ 13 Nov 2019
cugaaucccg cggacgaccc aucucggggc cguuuggguc ucuaccgucc ccuucuucau 1500
cugccguucc ggccgaccac ggggcgcacc ucucuuuacg cggucucccc gucugugccu 1560
ucucaucugc cggaccgugu gcacuucgcu ucaccucugc acgucgcaug gagaccaccg 1620
ugaacgccca ccaggucuug cccaaggucu uauauaagag gacucuugga cucucagcaa 1680
ugucaacgac cgaccuugag gcauacuuca aagacuguuu guuuaaggac ugggaggagu 1740 2019264591
ugggggagga gauuagguua augaucuuug uacuaggagg cuguaggcau aaauuggucu 1800
guucaccagc accaugcaac uuuuucaccu cugccuaauc aucucauguu cauguccuac 1860
uguucaagcc uccaagcugu gccuugggug gcuuuaggac auggacauug acccauauaa 1920
agaauuugga gcuucugugg aguuacucuc uuuuuugccu ucugacuuuu uuccuucuau 1980
ucgagaucuc cucgacaccg ccucugcucu guaucgggag gccuuagagu cuccggaaca 2040
uuguucaccu caccauacag cacucagaca agccauucug uguuggggug aguugaugaa 2100
ucuggccacc ugggugggaa guaauuugga agacccagca uccagggaau uaguagucag 2160
cuaugucaau guuaauaugg gccuaaaaau cagacaacua cugugguuuc acauuuccug 2220
ucuuacuuuu ggaagagaaa cuguucuuga guauuuggug ucuuuuggag uguggauucg 2280
cacuccuccu gcuuacagac caucaaaugc cccuaucuua ucaacacuuc cggaaacuac 2340
uguuguuaga cgacgaggca gguccccuag aagaagaacu cccucgccuc gcagacgaag 2400
gucucaaucg ccgcgucgca gaagaucuca aucucgggaa ccucaauguu aguaucccuu 2460
ggacucauaa ggugggaaac uuuacugggc uuuauucuuc uacuguaccu gucuuuaauc 2520
cugaguggca aacucccucu uuuccucaua uucauuugca ggaggacauu auuaauagau 2580
gucaacaaua ugugggcccu cuuacaguua augaaaaaag gagauuaaaa uuaauuaugc 2640
cugcuagguu cuauccuaac cuuaccaaau auuugcccuu ggacaaaggc auuaaaccau 2700
auuauccgga acaugcaguu aaucauuacu ucaaaacuag gcauuauuua cauacucugu 2760
ggaaggcngg cauucuauau aagagagaaa cuacacgcag cgccucauuu ugugggucac 2820
cauauucuug ggaacaagag cuacagcaug ggagguuggu cuuccaaacc ucgacaaggc 2880
auggggacaa aucuuucugu ucccaauccu cugggauucu uucccgauca ccaguuggac 2940
ccugcguucg gagccaacuc aaacaaucca gauugggacu ucaaccccaa caaggaucac 3000
Page 489
SIRIFD00001WOPCTSEQ 13 Nov 2019
uggccagagg caaaucaggu aggagcggga gcauucgggc caggguucac cccaccacac 3060
ggcggucuuu ugggguggag cccucaggcu cagggcacau ugacaacagu gccaguagca 3120
ccuccuccug ccuccaccaa ucggcaguca ggaagacagc cuacucccau cucuccaccu 3180
cuaagagaca gucauccuca ggccaugcag uggaa 3215
<210> 515 2019264591
<211> 3215 <212> RNA <213> Hepatitis B virus
<400> 515 aacuccacca cuuuccacca aacucuucaa gaucccagag ucagggcccu guacuuuccu 60
gcugguggcu ccaguucagg aacagugagc ccugcucuga auacugucuc ugccauaucg 120
ucaaucuuau cgaagacugg ggacccugua ccgaacaugg agaacaucgc aucaggacuc 180
cuaggacccc ugcucguguu acaggcgggg uuuuucuugu ugacaaaaau ccucacaaua 240
ccacagaguc uagacucgug guggacuucu cucaauuuuc uagggggaac acccgugugu 300
cuuggccaaa auucgcaguc ccaaaucucc agucacucac caaccuguug uccuccaauu 360
uguccugguu aucgcuggau gugucugcgg cguuuuauca ucuuccucug cauccugcug 420
cuaugccuca ucuucuuguu gguucuucug gacuaucaag guauguugcc cguuuguccu 480
cuaauuccag gaucaucaac aaccagcacc ggaccaugca aaaccugcac aacuccugcu 540
caaggaaccu cuauguuucc cucauguugc uguacaaaac cuacggacgg aaacugcacc 600
uguauuccca ucccaucauc uugggcuuuc gcaaaauacc uaugggagug ggccucaguc 660
cguuucucuu ggcucaguuu acuagugcca uuuguucagu gguucguagg gcuuuccccc 720
acugucuggc uuucaguuau auggaugaug ugguuuuggg ggccaagucu guacaacauc 780
uugagucccu uuaugccgcu guuaccaauu uucuuuuguc uuuggguaua cauuuaaacc 840
cucacaaaac aaaacgaugg ggauauuccc uuaacuucau gggauaugua auugggaguu 900
ggggcacauu gccacaggaa cauauuguac aaaaaaucaa acuguguuuu aggaaacuuc 960
cuguaaacag gccuauugau uggaaaguau gucaacgaau ugugggucua uugggguuug 1020
ccgccccuuu uacacaaugu ggauauccug cucuaaugcc uuuauaugca uguauacaag 1080
caaaacaggc uuuuacuuuc ucgccaacuu acaaggccuu ucuaaguaaa caguaucuga 1140 Page 490
SIRIFD00001WOPCTSEQ 13 Nov 2019
accuuuaccc cguugcucgg caacggccug gucugugcca aguguuugcu gacgcaaccc 1200
ccacugguug gggcuuggcg auaggccauc agcgcaugcg ugggaccuuu cugucuccuc 1260
ugccgaucca uacugcggaa cuccuagcag cuuguuuugc ucgcagcagg ucuggggcaa 1320
aacucaucgg gacugacaau ucugucguac ucucccgcaa guauacauca uuuccauggc 1380
ugcuaggcug ugcugccaac uggauccugc gcgggacguc cuuuguuuac gucccgucgg 1440 2019264591
cgcugaaucc cgcggacgac cccucgcggg gccgcuuggg gcucuaccgc ccgcuucucc 1500
gccuguucua ccgaccgacc acggggcgca ccucucuuua cgcggacucc ccgucugugc 1560
cuucucaucu gccggaccgu gugcacuucg cuucaccucu gcacgucgca uggagaccac 1620
cgugaacgcc cacaggaacc ugcccaaggu cuugcauaag aggacucuug gacuuucagc 1680
aaugucaacg accgaccuug aggcauacuu caaagacugu guguuuaaug agugggagga 1740
guugggggag gagguuaggu uaaaggucuu uguacuagga ggcuguaggc auaaauuggu 1800
guguucacca gcaccaugca acuuuuucac cucugccuaa ucaucucaug uucauguccu 1860
acuguucaag ccuccaagcu gugccuuggg uggcuuuggg gcauggacau ugacccguau 1920
aaagaauuug gagcuucugu ggaguuacuc ucuuuuuugc cuucugacuu cuuuccuucu 1980
auucgagauc uccucgacac cgccucugcu cuguaucggg aggccuuaga gucuccggaa 2040
cauuguucac cucaccauac ggcacucagg caagcuauuc uguguugggg ugaguugaug 2100
aaucuagcca ccuggguggg aaguaauuug gaagauccau cauccaggga auuaguaguc 2160
agcuauguca acguuaauau gggcauaaaa aucagacaac uauugugguu ucacauuucc 2220
ugucucacuu uugggagaga aacuguucuu gaauauuugg ugucuuuugg aguguggauu 2280
cgcacuccuc cugcauauag accaccaaau gccccuaucu uaucaacacu uccggaaacu 2340
acuguuguua gacgaagagg cagguccccu agaagaagaa cucccucgcc ucgcagacga 2400
aggucucaau cgccgcgucg cagaagaucu caaucucggg aaucucaaug uuaguauucc 2460
uuggacacau aaggugggaa acuuuacggg gcuuuauucu ucuacuguac cuugcuuuaa 2520
uccuaaaggg caaacuccuu cuuuuccuga uauucauuug caggaggaca uuguugauag 2580
auguaagcaa uuuguggggc cccuuacagu aaaugaaaac aggagacuaa aauuaauuau 2640
gccugcuaga uuuuauccca auguuacuaa auauuugccc uuagauaaag ggaucaaacc 2700 Page 491
SIRIFD00001WOPCTSEQ 13 Nov 2019
guauuaucca gaguauguag uugaucauua cuuccagacg cgacauuauu uacacacucu 2760
uuggaaggcg gggauuuuau auaaaagaga guccacacgu agcgccucau uuugcggguc 2820
accauauucu ugggaacaag aucuacagca ugggagguug gucuuccaaa ccucgaaaag 2880
gcauggggac aaaucuuucu guccccaauc cccugggauu cuuccccgau caucaguugg 2940
acccugcauu caaagccaac ucagaaaauc cagauuggga ccucaacccg cacaaggaca 3000 2019264591
acuggccgga cgccaacaag gugggagugg gagcauucgg gccgggguuc accccucccc 3060
augggggacu guuggggugg agcccucagg cucagggccu acucacaacu gugccaacag 3120
cgccuccucc ugccuccacc aaucggcagu caggaaggca gccuacuccc uuaucuccac 3180
cucuaaggga cacucauccu caggccgugc agugg 3215
<210> 516 <211> 3221 <212> RNA <213> Hepatitis B virus
<400> 516 auggcugcua gguuguacug ccaacuggau ucuucgaggg acguccuuug uuuacguccc 60
gucggcgcug aaucccgcgg acgaccccuc gcgaggccgc uuggggcugu aucguccccu 120
ucuccgucug ccguaccguc cgaccacggg gcgcaccucu cuuuacgcgg ucuccccguc 180
ugugccuucu caucugccgg uccgugugca cuucgcuuca ccucugcacg uugcauggag 240
accaccguga acgcccauca gguccugccc aaggucuuac auaagaggac ucuuggacuc 300
ucagcaaugu caacgaccga ccuugaggcc uacuucaaag acuguguguu uaaagacugg 360
gaggaguugg gggaggagau uagguuaaag gucuuuguau uaggaggcug uaggcauaaa 420
uuggucugcg caccaucauc augcaacuuu uucaccucug ccuaaucauc ucuuguacau 480
gucccacuuu ucaagccucc aagcugugcc uuggauggcu uuggggcaug gacauugacc 540
cuuauaaaga auuuggagcu acuguggagu uacucucauu uuugccuucu gacuucuuuc 600
cuuccguccg ggaucuacua gauacagccu cagcucuaua ucgggaagcc uuagagucuc 660
cugagcauug cucaccucac cauacagcac ucaggcaagc cauucucugc uggggggaau 720
uaaugacucu agcuaccugg guggguaaua auuuggaaga uccagcaucc agggaucuag 780
Page 492
SIRIFD00001WOPCTSEQ 13 Nov 2019
uagucaauua uguuaauacu aacaugggcc uaaagaucag gcaauuauug ugguuucaua 840
uuucuugccu uacuuuugga agagaaacug uccuugagua uuuggucucu uucggagugu 900
ggauucgcac uccaccagcu uauagaccac caaaugcccc uaucuuauca acacuuccgg 960
aaacuacugu uguuagacga cgagaccgag gcaggucccc uagaagaaga acucccucgc 1020
cucgcagacg aagaucucaa ucgccgcguc gcagaagauc ucaaucucgg gaaucucaau 1080 2019264591
guuaguauuc cuuggacuca uaagguggga aauuuuacug ggcuuuacuc uucuacuguc 1140
ccuaucuuua auccugaaug gcaaaccccu ucuuuuccua aaauucauuu acaugaagac 1200
auugcuaaua ggugucagca auuuguaggc ccucucacug uaaaugaaaa aagaagacug 1260
aaauuaauua ugccugcuag guuuuauccc aacagcacaa aauauuugcc uuuagacaaa 1320
gggauuaaac cuuauuaucc ugaucaugua guuaaucauu acuuucaaac ccgacauuau 1380
uuacauacuc uuuggaaggc ugggauucua uauaagaggg aaacuacacg uagcgccuca 1440
uuuugcgggu caccauauuc uugggaacaa gagcuacauc augggagguu ggucaucaaa 1500
accucgcaaa ggcaugggga cgaaccuuuc uguucccaac ccucugggau ucuuucccga 1560
ucaucaguug gacccugcau ucggagccaa uucaaacaau ccagauuggg acuucaaccc 1620
caucaaggac cacuggccac aagccaacca gguaggagug ggagcauucg ggccaggguu 1680
cacuccccca cacggaggug uuuuggggug gagcccucag gcucagggca uauuggccac 1740
agugccagca gugccuccuc cugccuccac caaucggcag ucaggaaggc agccuacucc 1800
caucucucca ccucuaagag acagucaucc ucaggccaug caguggaauu ccacagcuuu 1860
ccaccaagcu cugcaagauc ccagagucag gggccuguau uuuccugcug guggcuccag 1920
uucaggaaca cucaacccug uuccaacuau ugccucucac aucucgucaa ucuccucgag 1980
gauuggggac ccugugccga acauggagaa caucacauca ggauuccuag gaccccugcu 2040
cguguuacag gcgggguuuu ucuuguugac aagaauccuc acaauaccgc agagucuaga 2100
cucguggugg acuucucuca auuuucuagg gggaucaccc gugugucuug gccaaaauuc 2160
gcagucccca accuccaauc acucaccaac cuccuguccu ccaauuuguc cugguuaucg 2220
cuggaugugu cugcggcguu uuaucauauu ccucuucauc cugcugcuau gccucaucuu 2280
cuuauugguu cuucuggauu aucaagguau guugcccguu uguccucuaa uuccaggauc 2340
Page 493
SIRIFD00001WOPCTSEQ 13 Nov 2019
aacaacaacc agcacgggac ccugcaaaac cugcacgacu ccugcucaag gcaacucuau 2400
guuucccuca uguugcugua caaaaccuac ggauggaaau ugcaccugua uucccauccc 2460
aucaucuugg gcuuucgcaa aauaccuaug ggagugggcc ucaguccguu ucucuuggcu 2520
caguuuacua gugccauuug uucagugguu cguagggcuu ucccccacug uuuggcuuuc 2580
agcuauaugg augauguggu acugggggcc aagucuguac agcaucuuga gucccuuuau 2640 2019264591
accgguguua ccaauuuucu uuugucuuug gguauacauu uaaacccuaa caaaacaaag 2700
agaugggguu auucacugaa uuucaugggu uauguaauug gaaguugggg uacauugcca 2760
caggaucaua uuguacaaaa aaucaaacac uguuuuagaa aacuuccugu caaucgaccu 2820
auugauugga aaguauguca aagaauugug ggucuuuugg gcuuugccgc uccauuuaca 2880
caaugugguu auccugcauu aaugccuuug uaugcaugua uacaagcaaa acaggcuuuu 2940
acuuucucgc caacuuacaa ggccuuucua aguaaacagu auaugaaccu uuaccccguu 3000
gcccggcaac ggccuggucu gugccaagug uuugcugacg caacccccac uggcuggggc 3060
uuggccaucg gccaucagcg caugcgcgga accuuugugg cuccucugcc gauccauacu 3120
gcggaacucc uggccgcuug uuuugcucgc agccggucug gagcaaaacu caucgggacu 3180
gauaauucug ucguccuuuc ucggaaauau acaucauuuc c 3221
<210> 517 <211> 3215 <212> RNA <213> Hepatitis B virus
<400> 517 cuccaccacg uuccaccaaa cucuucaaga ucccagaguc agggcucugu acuuuccugc 60
ugguggcucc aguucaggaa caguaaaccc uguucagaac acugccucuu ccauaucguc 120
aaucuuaucg acgacugggg acccugugcc gaacauggag aacaucgcau caggacuccu 180
aggaccccug cucguguuac aggcgggguu uuucucguug acaaaaaucc ucacaauacc 240
acagagucua gacucguggu ggacuucucu caauuuucua gggggaacac ccgugugucu 300
uggccaaaau ucgcaguccc aaaucuccag ucacucacca acuuguuguc cuccgauuug 360
uccugguuau cgcuggaugu gucugcggcg uuuuaucauc uuccucugca uccugcugcu 420
augccucauc uucuuguugg uucuucugga cuaucaaggu auguugcccg uuuguccucu 480 Page 494
SIRIFD00001WOPCTSEQ 13 Nov 2019
aauuccagga ucaucaacca ccagcacagg accaugcaaa accugcacga cuccugcuca 540
aggaaccucu auguuucccu cauguugcug uauaaaaccu acggacggaa acugcaccug 600
uauucccauc ccaucaucuu gggcuuucgc aaaauaccua ugggaguggg ccucaguccg 660
uuucucuugg cucaguuuac uagugccauu uguucagugg uucguagggc uuucccccac 720
ugucuggcuu ucaguuauau ggaugaugug guuuuggggg ccaagucugu acaacaucuu 780 2019264591
gagucccuuu augccgcugu uaccaauuuu cuuuugucuu ugggcauaca uuuaaacccu 840
cagaaaacaa aaagaugggg cuacucccuu aacuucaugg gguauguaau uggaaguugg 900
gggaccuuac cccaagaaca uauuguguug aaaaucaaac aauguuuuag gaaacuuccu 960
guaaacaggc cuauugauug gaaaguaugu caacgaauug ugggucuuuu gggauuugcu 1020
gcuccuuuca cacaaugugg auauccugcu uuaaugccuu uauaugcaug uauacaagcu 1080
aaacaggcuu uuacuuuuuc gccaacguau aaggccuuuc uccacaaaca auaucugaac 1140
cuuuaccccg uugcucggca acggccaggu cugugccaag uguuugcuga cgcaaccccc 1200
acuggcuggg gcuuggccau aggccaucag cgcaugcgug ggaccuuugu gucuccucug 1260
ccgauccaua cugcggaacu ccuagccgcu uguuuugcuc gcagcagguc uggagcaaaa 1320
cuuaucggga cugacaauuc ugucguccuu ucccgcaaau auacaucguu uccauggcug 1380
cuaggcugug cugccaacug gauccugcgc gggacguccu uugucuacgu cccgucggcg 1440
cugaaucccg cggacgaccc cucccggggc cgcuuggggc ucuaccgccc gcuucuccgc 1500
cugccguacc guccgaccac ggggcgcacc ucucuuuacg cggacucccc gucugugccu 1560
ucucaucugc cggaccgugu gcacuucgcu ucaccucugc acgucgcaug gagaccaccg 1620
ugaacgccca ucggaaccug cccaaggucu ugcauaagag gacucuugga cuuucagcaa 1680
ugucaacgac cgaccuugag gcauacuuca aagacugugu guuuacugag ugggaggagu 1740
ugggggagga gaucagguua aaggucuuug uacuaggagg cuguaggcau aaauuggucu 1800
guucaccagc accaugcaac uuuuucaccu cugccuaauc aucucauguu cauguccuac 1860
uguucaagcc uccaagcugu gccuugggug gcuuuggggc auggacauug acaccuauaa 1920
agaauuugga gcuucugugg aguuacucuc uuuuuugccu ucugacuucu uuccuucuau 1980
ucgagaucuu cucgacaccg ccucugcucu guaucgggag gccuuagagu cuccggaaca 2040 Page 495
SIRIFD00001WOPCTSEQ 13 Nov 2019
uuguucaccu caccauacgg cacucaggca agcuauugug uguuggggug aguugaugaa 2100
ucuagccacc ugggugggaa guaauuugga agacccagcc ucccgggaau uaguagucag 2160
uuaugucaau guuaauaugg gccuaaaaau cagacaacua uugugguuuc acauuuccug 2220
ucuuacuuuu ggaagagaaa cuguucuuga auauuuggug ucuuuuggag uguggauucg 2280
cacaccucca gcauauagac caccaaaugc cccuaucuua ucaacacuuc cggaaacuac 2340 2019264591
uguuguuaga cgacgaggca gguccccuag aagaagaacu cccucgccuc gcagacgaag 2400
gucucaaucg ccgcgucgca gaagaucuca aucucgggaa ucuaaauguu aguauuccuu 2460
ggacucauaa ggugggaaac uuuacggggc uuuauucuuc uacgguaccu agcuuuaauc 2520
cucaauggca aacuccuuca uuuccugaca uucauuugca ggaggacauc auugauaagu 2580
guaaacaauu ugugggaccc cuuacaguga augaaaacag gagacuaaaa uugauuaugc 2640
cugcuagguu cuaucccaau guuacuaaau auuugcccuu agauaaagga auuaaaccuu 2700
auuauccaga gcauguaguu aaucauuacu uccagacgag acauuauuua cauacucuuu 2760
ggaaggcggg uaucuuauau aaaagagaga caacacguag cgccucauuu ugcgggucac 2820
cauauucuug ggaacaagag cuacagcaug ggagguuggu ccuccaaacc ucgaaaaggc 2880
auggggacaa aucuuuccgu ccccaauccu cugggauucu uucccgauca ccaguuggac 2940
ccugcauuca aagccaacuc cgacaauccc gauugggacc ucaacccaca caaggacaac 3000
uggccggacu ccaacaaggu gggaguggga gcauucgggc cgggauucac uccaccccau 3060
gggggacugu ugggguggag cccucaagcu cagggcauac ucacaacugu gccaacagcu 3120
ccuccuccug ccuccaccaa ucggcaguua ggaaggaagc cuacuccccu gucuccaccu 3180
cuaagagaca cucauccuca ggcaaugcag uggaa 3215
<210> 518 <211> 3212 <212> RNA <213> Hepatitis B virus
<400> 518 uuccacaaca uuucaccaag cucugcagga ucccagagua agaggccugu auuuuccugc 60
ugguggcucc aguuccggaa cagugaaccc uguuccgacu acugccucac ucaucucguc 120
Page 496
SIRIFD00001WOPCTSEQ 13 Nov 2019
aaucuucucg aggauugggg acccugcacc gaacauggaa agcaucacau caggauuccu 180
aggaccccug cucguauuac aggcgggguu uuucuuguug acaaaaaucc ucacaauacc 240
gcagagucua gacucguggu ggacuucucu caauuuucua gggggagcuc ccgugugucu 300
uggccaaaau ucgcaguccc caaccuccaa ucacucacca accucuuguc cuccaauuug 360
uccuggcuau cgcuggaugu gucugcggcg uuuuaucauc uuccucuuca uccugcugcu 420 2019264591
augccucauc uucuuguugg uucuucugga cuaucaaggu auguugcccg uuuguccucu 480
aauuccagga ucaucaacca ccaguacggg acccugccga accugcacga cucuugcuca 540
aggaaccucu auguuucccu cauguugcug uucaaaaccu ucggacggaa auugcacuug 600
uauucccauc ccaucaucau gggcuuucgg aaaauuccua ugggaguggg ccucagcccg 660
uuucuccugg cucaguuuac uagugccauu uguucagugg uucgccgggc uuucccccac 720
ugucuggcuu ucaguuauau ggaugaugug guauuggggg ccaagucugu acaacaucuu 780
gagucccuuu auaccucugu uaccaauuuu cuuuugucuu uggguauaca uuuaaauccc 840
aacaaaacaa aaagaugggg auauucccug aauuucaugg guuauguaau uggaaguugg 900
ggaucauuac cacaggaaca caucauaaug aaaaucaaag acuguuuuag aaaacucccc 960
guuaaccggc cuauugauug gaaaguaugu caacgaauug ugggucuuuu gggcuuugcu 1020
gccccuuuua cacaaugugg guauccugcu uuaaugccuc uguaugcgug uauucaaucu 1080
aagcaggcuu ucacuuucuc gccaacuuac aaggccuuuc uguguaaaca auaccugaac 1140
cuuuaccccg uugcccggca acggccaggu cugugccaag uguuugcuga ugcaaccccc 1200
acuggcuggg gcuuggccau aggccaucag cgcaugcgcg gaaccuuuau ggcuccucug 1260
ccgauccaua cugcggaacu ccuagccgcu uguuuugcuc gcagcagguc uggagcgaaa 1320
cuuaucggga cagauaauuc ugucguucuc ucccggaaau auacauccuu uccauggcug 1380
cuaggcugug cugccaacug gauccugcga gggacguccu uugucuacgu cccgucagcg 1440
cugaauccug cggacgaccc gucucggggu cgcuugggga ucuaucgucc ccuucuccgu 1500
cugccguucc agccguccac ggggcgcacc ucucuuuacg cggucucccc gucugugccu 1560
ucucaucugc cggaccgugu gcacuucgcu ucaccucugc acgucgcaug gagaccaccg 1620
ugaacgccca ccaaaucuug cccaaggucu uacauaagag gacucuugga cucucugcaa 1680
Page 497
SIRIFD00001WOPCTSEQ 13 Nov 2019
ugucaacgac cgaccuugag gcauacuuca aagacuguuu guuuaaagac ugggaggagu 1740
ugggggagga gauuagauua aaggucuuug uacuaggagg cuguaggcau aaauuggucu 1800
gcgcaccagc accaugcaac uuuuucaccu cugccuaauc aucucuuguu cauguccuac 1860
uguucaagcc uccaagcugu gccuugggug gcuuuggggc auggacauug acccuuauaa 1920
agaauuugga gcuacugugg aguuacucuc guuuuugccu ucugacuucu uuccuucagu 1980 2019264591
aagagaucuu cuagauaccg ccucagcuuu guaucgggau gccuuagaau cuccugagca 2040
uuguucaccg caucacacug cacucaggca agccauucuu ugcugggggg aacuaaugac 2100
ucuagcuacc ugggugggug uaaauuugga agauccagca uccagggacc uaguagucag 2160
uuaugucaau acuaauaugg gccuaaaguu caggcaauua uugugguuuc acauuucuug 2220
ucucacuuuu ggaagagaaa ccgucauaga guauuuggug ucuuuuggag uguggauucg 2280
cacuccucca gcuuauagac caccaaaugc cccuaucuua ucaacacuuc cggagaauac 2340
uguuguuaga cgaagaggca gguccccuag aagaagaacu cccucgccuc gcagacgaag 2400
aucucaaucg ccgcgucgca gaagaucuca aucuccagcu ucccaauguu aguauuccuu 2460
ggacucacaa ggugggaaau uuuacggggc uuuacucuuc uacuauaccu gucuuuaauc 2520
cuaacuggaa aacuccaucu uuuccugaua uucauuugca ccaggacauu auuaacaaau 2580
gugaacaauu uguagguccu cuaacaguaa augaaaaacg aagauuaaac uuagucaugc 2640
cugcuagauu uuuucccauc ucuacgaaau auuugccccu agagaaaggu auaaaaccuu 2700
auuauccaga uaauguaguu aaucauuacu uccaaaccag acacuauuua cacacccuau 2760
ggaaggcggg caucuuauau aaaagagaaa cuacacguag cgccucauuu ugugggucac 2820
cuuauucuug ggaacaagag cuacaucaug gggcuuucuu ggacgguccc ucucgaaugg 2880
gggaagaauc auuccaccac caauccucug ggauuuuuuc ccgaccacca guuggaucca 2940
gcauucagag caaacaccag aaauccagau ugggaccaca aucccaacaa agaccacugg 3000
acggaagcca acaagguagg agugggagca uucgggccgg gguucacucc cccacacgga 3060
ggccuuuugg gguggagccc ucaggcucaa ggcaugcuaa aaacauugcc agcagauccg 3120
ccuccugccu ccaccaaucg gcagucagga aggcagccua ccccaaucac uccaccuuug 3180
agagacacuc auccucaggc caugcagugg aa 3212
Page 498
SIRIFD00001WOPCTSEQ 13 Nov 2019
<210> 519 <211> 3248 <212> RNA <213> Hepatitis B virus
<400> 519 cucuacagca uuccaccaag cucuacaaaa ucccaaaguc aggggccugu auuuuccugc 60
ugguggcucc aguucaggga uagugaaccc uguuccgacu auugccucuc acaucucguc 120 2019264591
aaucuucucc aggauugggg acccugcacc gaacauggag aacaucacau caggauuccu 180
aggaccccug cucguguuac aggcgggguu uuucuuguug acaagaaucc ucacaauacc 240
gcagagucua gacucguggu ggacuucucu caauuuucua gggggagugc ccgugugucc 300
uggccuaaau ucgcaguccc caaccuccaa ucacucacca aucuccuguc cuccaacuug 360
uccuggcuau cgcuggaugu gucugcggcg uuuuaucaua uuccucuuca uccugcugcu 420
augccucauc uucuuguugg uucuucugga cuaucaaggu auguugcccg uuuguccucu 480
gauuccagga uccucgacca ccaguacggg acccugcaaa accugcacga cuccugcuca 540
aggcaacucu auguaucccu cauguugcug uacaaaaccu ucggacggaa auugcaccug 600
uauucccauc ccaucaucuu gggcuuucgc aaaauaccua ugggaguggg ccucaguccg 660
uuucucuugg cucaguuuac uagugccauu uguucagugg uucguagggc uuucccccac 720
ugucuggcuu ucagcuauau ggaugaugug guauuggggg ccaaaucugu acaacaucuu 780
gagucccuuu auaccgcugu uaccaauuuu cuuuugucuu uggguauaca ucuaaacccu 840
aacaaaacaa aaagaugggg uuauuccuua aauuuuaugg gauauguaau uggaaguugg 900
gguacuuugc cacaagaaca caucacacag aaaauuaagc aauguuuucg gaaacucccu 960
guuaacaggc caauugauug gaaagucugu caacgaauaa cuggucuguu ggguuucgcu 1020
gcuccuuuca cccaaugugg uuacccugcc uuaaugccuu uauaugcaug uauacaagcu 1080
aagcaggcuu uuacuuucuc gccaacuuau aaggccuuuc ucuguaaaca auacaugaac 1140
cuuuaccccg uugcuaggca acggcccggu cugugccaag uguuugcuga cgcaaccccc 1200
acugguuggg gcuuggccau cggccaucag cgcaugcgug gaaccuuugu ggcuccucug 1260
ccgauccaua cugcggaacu ccuagcugcu uguuuugcuc gcagccgguc uggagcaaaa 1320
cucauuggga cugacaauuc ugucguccuu ucucggaaau auacauccuu uccauggcug 1380 Page 499
SIRIFD00001WOPCTSEQ 13 Nov 2019
cuaggcugug cugccaacug gauccuucgc gggacguccu uuguuuacgu cccgucagcg 1440
cugaauccag cggacgaccc cucccggggc cguuuggggc ucugucgccc ccuucuccgu 1500
cugccguucc ugccgaccac ggggcgcacc ucucuuuacg cggucucccc gucuguuccu 1560
ucucaucugc cggaccgugu gcacuucgcu ucaccucugc acguuacaug gaaaccgcca 1620
ugaacaccuc ucaucaucug ccaaggcagu uauauaagug gacucuugga cuguuuguua 1680 2019264591
ugucaacaac cgggguggag aaauacuuca aggacugugu uuuugcugag ugggaagaau 1740
uaggcaauga guccagguua augaccuuug uauuaggagg cuguaggcau aaauuggucu 1800
gcgcaccagc accauguaac uuuuucaccu cugccuaauc aucucuuguu cauguccuac 1860
uguucaagcc uccaagcugu gccuugggug gcuuuagggc auggauagaa caacuuugcc 1920
auauggccuu uuuggcuuag acauugaccc uuauaaagaa uuuggagcua cuguggaguu 1980
gcucucguuu uugccuucug acuuuuuccc gucuguucgu gaucuucucg acaccgcuuc 2040
agcuuuguac cgggaauccu uagaguccuc ugaucauugu ucgccucacc auacagcacu 2100
caggcaagca auccugugcu ggggugaguu gaugacucua gcuaccuggg uggguaauaa 2160
uuuggaagau ccagcaucca gagauuuggu ggucaauuau guuaauacua auauggguuu 2220
aaaaaucagg caacuauugu gguuucacau uuccugucuu acuuuuggga gagaaaccgu 2280
ucuugaguau uuggugucuu uuggagugug gauucgcacu ccuccugcuu auagaccacc 2340
aaaugccccu auccuaucaa cacuuccgga gacuacuguu guuagacgaa gaggcagguc 2400
cccucgaaga agaacucccu cgccucgcag acgaagaucu caaucgccgc gucgcagaag 2460
aucugcaucu ccagcuuccc aauguuagua uuccuuggac ucacaaggug ggaaacuuua 2520
cggggcugua uucuucuacu auaccugucu uuaauccuga uuggcaaacu ccuucuuuuc 2580
caaauaucca uuugcaucaa gacauuauaa cuaaauguga acaauuugug ggcccucuca 2640
caguaaauga gaaacgaaga uuaaaacuag uuaugccugc cagauuuuuc ccaaacucua 2700
cuaaauauuu accauuagac aaagguauca aaccguauua uccagaaaau guaguuaauc 2760
auuacuucca gaccagacau uauuuacaua cccuuuggaa ggcggguauu cuauauaaga 2820
gagaaacauc ccguagcgcu ucauuuugug ggucaccaua uacuugggaa caagaucuac 2880
agcauggggc uuucuuggac ggucccucuc gaguggggaa agaaccuuuc cgccagcaau 2940 Page 500
SIRIFD00001WOPCTSEQ 13 Nov 2019
ccucuaggau uccuucccga ucaccaguug gacccagcau ucagagcaaa uaccaacaau 3000
ccagauuggg acuucaaucc caaaaaggac ccuuggccag aggccaacaa gguaggaguu 3060
ggagccuaug gacccggguu caccccucca cacggaggcc uuuuggggug gagcccucag 3120
ucucagggca cacuaacaac uuugccagca gauccgccuc cugccuccac caaucgucag 3180
ucagggaggc agccuacucc caucucucca ccacuaagag acagucaucc ucaggccaug 3240 2019264591
caguggaa 3248
<210> 520 <211> 3215 <212> RNA <213> Hepatitis B virus
<400> 520 cucaacucag uuccaccagg cucuguugga uccgagggua agggcucugu auuuuccugc 60
ugguggcucc aguucaggga cacagaaccc ugcuccgacu auugccucuc ucacaucauc 120
aaccuucucg aagacugggg gcccugccau gaacauggag aacaucacau caggacuccu 180
aggaccccug cucguguuac aggcggugug uuucuuguug acaaaaaucc ucacaauacc 240
acagagucua gacucguggu ggacuucucu caauuuucua gggggacuac ccgggugucc 300
uggccaaaau ucgcaguccc caaccuccaa ucacuuacca accuccuguc cuccaacuug 360
uccuggcuau cguuggaugu gucugcggcg uuuuaucauc uuccucuuca uccugcugcu 420
augccucauc uucuuguugg uucuucugga cuaccaaggu auguugcccg uuuguccucu 480
acuuccagga uccacgacca ccagcacggg accaugcaaa accugcacaa cucuugcuca 540
aggaaccucu auguuucccu ccuguugcug uuccaaaccc ucggacggaa acugcaccug 600
uauucccauc ccaucaucuu gggcuuuagg aaaauaccua ugggaguggg ccucagcccg 660
uuucuccugg cucaguuuac uagugcaauu uguucagugg ugcguagggc uuucccccac 720
ugucuggcuu uuaguuauau ggaugaucug guauuggggg ccaaaucugu gcagcaucuu 780
gagucccuuu auaccgcugu uaccaauuuu cuguuaucug uggguaucca uuuaaauacu 840
gcuaaaacaa aaagaugggg uuacacccua cauuucaugg guuauguuau ugguaguugg 900
ggaacguuac cccaagacca uauuauacac aaaaucaaag auuguuuucg gaaacuuccu 960
Page 501
SIRIFD00001WOPCTSEQ 13 Nov 2019
guaaaucguc caauugauug gaaaguuugu caacgcauug ugggucuuuu gggcuuugcg 1020
gccccuuuca cccaaugugg uuauccugcu cucaugccuu uguauaccug uauuacugcu 1080
aaacaggcuu uugucuuuuc gccaacuuac aaggccuuuc ucuguaaaca auacaugaac 1140
cuuuaccccg uugcucggca acggccaggc cugugccaag uguuugcuga cgcaaccccc 1200
acugguuggg gcuuggccau uggccaucag cgcaugcgug gaaccuuugu ggcuccucug 1260 2019264591
ccgauccaua cugcggaacu ccuugcagcc uguuucgcuc gcagccgguc uggagcgaac 1320
auuaucggua cagacaacuc uguuguccuc ucuaggaagu acaccuccuu uccauggcug 1380
cucgguugug cugccaacug gauccugcgc gggacguccu uuguuuacgu cccgucggcg 1440
cugaaucccg cggacgaccc cucccggggu cgcuuggggc uguaccgccc ccuucuucgu 1500
cugccguucc agccgaccac gggucgcacc ucucuuuacg cggacucccc gucuguuccu 1560
ucucaucugc cggaccgugu gcacuucgcu ucaccucugc acgucgcaug gagaccaccg 1620
ugaacgcccc cuggaaucug ccaacagucu uacauaagag gacucuugga cuuucaggac 1680
ggucaaugac cuggaucgaa gacuacauca aagacugugu auuuaaggac ugggaggagc 1740
ugggggagga gaucagguua aaggucuuug uacuaggagg cuguaggcau aaauuggucu 1800
guucaccagc accaugcaac uuuuucaccu cugccuaauc aucuuuuguu caugucccac 1860
uguucaagcc uccaagcugu gccuugggug gcuuuggggc auggacauug acccuuauaa 1920
agaauuugga gcuucugugg aauugcucuc uuuuuugccu ucugauuucu ucccgucugu 1980
ucgggaccua cucgacaccg cuucagcccu guaccgggau gcucuagagu caccggaaca 2040
uugcaccccc aaucauaccg cucucaggca agcuauuuug ugcuggggug aguuaaugac 2100
uuuggcuucc ugggugggua auaauuugga agacccugca gcuagggauu uaguaguuaa 2160
uuaugucaac acuaauaugg gccugaaaau uagacaacug uugugguuuc acauuuccug 2220
ucuuacuuuu ggaagagaaa caguucuuga guauuugguc uccuuuggag uguggauucg 2280
cacuccaccu gcuuauagac caccaaaugc cccuauccua uccacacuuc cggaaacuac 2340
uguuguuaga cgacgaggca gguccccuag aagaagaacu cccucgccuc gccgacgaag 2400
gucucaaucg ccgcgucgca gaagaucuca aucuccagcu ucccaauguu aguauuccuu 2460
ggacucauaa ggugggaaau uuuacggggc ucuacucuuc uacuguaccu gcuuucaauc 2520
Page 502
SIRIFD00001WOPCTSEQ 13 Nov 2019
cuaacugguu aacuccuucu uuuccugaua uucauuuaca ccaggaucug auaucuaaau 2580
gugaacaauu uguaggcccg cucacuaaaa augaauugag aagauuaaaa uuggucaugc 2640
cagcuagauu uuauccuaag guuaccaaau acuuuccuau ggagaaaggg auuaaacccu 2700
auuauccuga gcaugcaguu aaucauuauu uuaaaacaag acauuauuug cauacuuuau 2760
ggaaggcggg aaucuuauau aagagagaau ccacacguag cgccucauuu ugugggucac 2820 2019264591
cauauuccug ggaacaagag cuacagcaug ggagcaccuc ucucaacgac aagaaggggc 2880
augggacuga aucuuucugu gcccaauccu cugggcuucu ugccagacca ucagcuggau 2940
ccgcuauuca gagcaaauuc cagcaguccc gacugggacu ucaacacaaa caaggacagu 3000
uggccaaugg caaacaaggu aggaguggga ggcuacgguc caggguucac acccccucac 3060
gguggccugc ugggguggag cccucaggca caggguguuu uaacaaccuu gccagcagau 3120
ccgccuccug cuuccaccaa ucggcggucc gggagaaagc caaccccagu cucuccaccu 3180
cuaagagaca cacauccaca ggccaugcag uggaa 3215
<210> 521 <211> 3215 <212> RNA <213> Hepatitis B virus
<400> 521 cucaacccag uuccaccagg ccuuguugga uccgagggua agggcucugu auuuuccugc 60
ugguggcucc aguucagaga cgcagaaccc ugcuccgacu auugccucuc ucacaucauc 120
aaucuucucg aagacugggg gcccugcuau gaacauggac agcaucacau caggacuccu 180
aggaccccug cucguguuac aggcggugug uuucuuguug acaaaaaucc ucacaauacc 240
acagagucua gacucguggu ggacuucucu caauuuucua gggggacuac ccgggugucc 300
uggccaaaau ucgcaguccc caaccuccaa ucacuuacca accuccuguc cuccaacuug 360
uccuggcuau cguuggaugu gucugcggcg uuuuaucauc uuccucuuca uccugcugcu 420
augccucauc uucuuguugg uucuucugga cuaucaaggu auguugcccg uuuguccucu 480
aauuccagga ucuacgacca ccagcacggg accaugcaaa accugcacaa cucuugcuca 540
aggaaccucu auguuucccu ccuguugcug uuccaaaccc ucggacggaa acugcaccug 600
uauucccauc ccaucaucuu gggcuuuagg aaaauaccua ugggaguggg ccucagcccg 660 Page 503
SIRIFD00001WOPCTSEQ 13 Nov 2019
uuucuccugg cucaguuuac uagugcaauu uguucagugg ugcguagggc uuucccccac 720
ugucuggcuu uuaguuauau ggaugaucug guauuggggg ccaaaucugu gcagcaucuu 780
gagucccuuu auaccgcugu uaccaauuuu cuguuaucug uggguaucca uuugaauacc 840
ucuaaaacaa aaagaugggg uuacaauuua cauuucaugg guuauguaau uggcaguugg 900
ggagcauuac cccaagauca uauuguacac aaaaucaaag aauguuuuag aaaacuuccu 960 2019264591
guaaaucguc caauugacug gaaaguuugu caacguauug ugggacuuuu gggcuuugcu 1020
gcuccuuuua cccaaugugg uuauccugcu cucaugccuc uguauaccug uaucacugcg 1080
aaacaggcuu uugucuuuuc gccaacuuac aaggccuuuc ucuguaaaca guacaugaac 1140
cuuuaccccg uugcucggca acggccaggc cugugccaag uguuugcuga cgcaaccccc 1200
acugguuggg gcuuggccau uggccaucag cgcaugcgug gaaccuuugu ggcuccucug 1260
ccgauccaua cugcggaacu ccuugcagcu uguuucgcuc gcagccgguc uggagcgaau 1320
aucaucggga cagacaacuc uguuguacuc ucuaggaagu acaccuccuu cccauggcug 1380
cucggaugug cugccaacug gauccugcgc gggacguccu uuguuuacgu cccgucggcg 1440
cugaaucccg cggacgaccc cucccggggc cgcuuggggc ucuaccgccc ucuucugcgu 1500
cugccguucc agccgaccac gggucgcacc ucucuuuacg cggacucccc gucuguuccu 1560
ucucaucugc cgguccgugu gcacuucgcu ucaccucugc acgucgcaug gagaccaccg 1620
ugaacgcccc cuggaguuug ccaacagucu uacauaaaag gacuauugga cuuucaggac 1680
ggucaaugac cuggaucgaa gaauacauca aagacugugu auuuaaagac ugggaggagu 1740
ugggggagga gaucagguua aaggucuuug uacuaggagg cuguaggcau aaauuggucu 1800
guucaccagc accaugcaac uuuuucaccu cugccuaauc aucuuuuguu cauguccuac 1860
uguucaagcc uccaagcugu gccuugggug gcuuuggggc auggacauug acccuuauaa 1920
agaauuugga gcuucugugg aauugcucuc uuuuuugccu ucugauuucu ucccgucggu 1980
ucgggaccua cucgacaccg cuucagcucu cuacagggau gcuuuagagu caccggaaca 2040
uugcaccccc aaucauaccg cucucaggca agcuauuuug uguuggggug aauuaaugac 2100
uuuggcuucc ugggugggca auaauuugga ggacccugca gccagggauu uaguaguuaa 2160
cuauguuaac acuaacaugg gcuuaaagau uagacaauua uugugguuuc acauuuccug 2220 Page 504
SIRIFD00001WOPCTSEQ 13 Nov 2019
ccuuacuuuu ggaagagaaa caguucuuga guauuuggug uccuuuggag uguggauucg 2280
cacuccucca gcuuauagac caccaaaugc cccuauccua uccacacuuc cggaaacuac 2340
uguuguuaga cgacgaggca gguccccuag aagaagaacu cccucgccuc gcagacgaag 2400
gucucaaucg ccgcgucgca gaagaucuca aucuccagcu ucccaauguu aguauuccuu 2460
ggacucauaa ggugggaaau uuuacggggc ucuacucuuc uacuguaccu gcuuucaauc 2520 2019264591
cucacugguu aacuccuucu uuuccugaua uucauuugca ucaagaccug auaucuaaau 2580
gugaacaauu uguaggccca cuuaccaaaa augaauugag aagguugaaa uugauuaugc 2640
cagccagauu cuuuccuaaa cuuacuaaau auuuuccucu ggagaaaggc auuaaaccuu 2700
auuauccuga gcaugcaguu aaucauuauu uuaagaccag acauuauuug cauacuuuau 2760
ggaaggcggg aauuuuauau aagagagaau ccacacguag cgccucauuu ugugggucac 2820
cauauuccug ggaacaagag cuacagcaug ggagcaccuc ucucaacgac aagaaggggc 2880
augggacaga aucucucugu gcccaaucca cugggauucu uuccagaaca ucagcuggau 2940
ccucuuuuca gagcaaauuc cagcaguccc gauugggacu ucaacaaaaa caaggacacu 3000
uggccaaugg caaacaaggu aggaguggga gguuacgguc caggguucac acccccacac 3060
gguggccugu ugggguggag cccucaggca caagguguuc uaacaaccuu gccagcagau 3120
ccgccuccug cuuccaccaa ucggcggucc gggaggaagc caaccccagu cucuccaccu 3180
cuaagagaca cacauccaca ggcaaugcag uggaa 3215
<210> 522 <211> 3248 <212> RNA <213> Hepatitis B virus
<400> 522 cucuacagca uuccaccaag cucuacaaaa ucccaaaguc aggggccugu auuuuccugc 60
ugguggcucc aguucaggga uagugaaccc uguuccgacu auugccucuc acaucucguc 120
aaucuucucc aggauugggg acccugcacc gaauauggag aacaucacau caggauuccu 180
aggaccccug cucguguuac aggcgggguu uuucuuguug acaagaaucc ucacaauacc 240
gcagagucua gacucguggu ggacuucucu caauuuucua gggggagugc ccgugugucc 300
Page 505
SIRIFD00001WOPCTSEQ 13 Nov 2019
uggccuaaau ucgcaguccc caaccuccaa ucacucacca aucuccuguc cuccaacuug 360
uccuggcuau cgcuggaugu gucugcggcg uuuuaucaua uuccucuuca uccugcugcu 420
augccucauc uucuuguugg uucuucugga cuaucaaggu auguugcccg uuuguccucu 480
gauuccagga uccucgacca ccaguacggg acccugcaaa accugcacga cuccugcuca 540
aggcaacucu auguaucccu cauguugcug uacaaaaccu ucggacggaa auugcaccug 600 2019264591
uauucccauc ccaucaucuu gggcuuucgc aaaauaccua ugggaguggg ccucaguccg 660
uuucucuugg cucaguuuac uagugccauu uguucagugg uucguagggc uuucccccac 720
ugucuggcuu ucagcuauau ggaugaugug guauuggggg ccaaaucugu acaacaucuu 780
gagucccuuu auaccgcugu uaccaauuuu cuuuugucuu uggguauaca ucuaaacccu 840
aacaaaacaa aaagaugggg uuauuccuua aauuuuaugg gauauguaau uggaaguugg 900
gguacuuugc cacaagaaca caucacacag aaaauuaagc aauguuuucg gaaacucccu 960
guuaacaggc caauugauug gaaagucugu caacgaauaa cuggucuguu ggguuucgcu 1020
gcuccuuuua cccaaugugg uuacccugcc uuaaugccuu uauaugcaug uauacaagcu 1080
aagcaggcuu uuacuuucuc gccaacuuau aaggccuuuc ucuguaaaca auacaugaac 1140
cuuuaccccg uugcuaggca acggcccggu cugugccaag uguuugcuga cgcaaccccc 1200
acugguuggg gcuuggccau cggccaucag cgcaugcgug gaaccuuugu ggcuccucug 1260
ccgauccaua cugcggaacu ccuagcugcu uguuuugcuc gcagccgguc uggagcaaaa 1320
cucauuggga cugacaauuc ugucguccuu ucucggaaau auacauccuu uccauggcug 1380
cuaggcugug cugccaacug gauccuucgc gggacguccu uuguuuacgu cccgucagcg 1440
cugaauccag cggacgaccc cucccggggc cguuuggggc ucugucgccc ccuucuccgu 1500
cugccguucc ugccgaccac ggggcgcacc ucucuuuacg cggucucccc gucuguuccu 1560
ucucaucugc cggaccgugu gcacuucgcu ucaccucugc acguuacaug gaaaccgcca 1620
ugaacaccuc ucaucaucug ccaaggcagu uauauaagag gacucuugga cuguuuguua 1680
ugucaacaac cgggguggag aaauacuuca aggacugugu uuuugcugag ugggaagaau 1740
uaggcaauga guccagguua augaccuuug uauuaggagg cuguaggcau aaauuggucu 1800
gcgcaccagc accauguaac uuuuucaccu cugccuaauc aucucuuguu cauguccuac 1860
Page 506
SIRIFD00001WOPCTSEQ 13 Nov 2019
uguucaagcc uccaagcugu gccuugggug gcuuuagggc auggauagaa caacuuugcc 1920
auauggccuu uuuggcuuag acauugaccc uuauaaagaa uuuggagcua cuguggaguu 1980
gcucucguuu uugccuucug acuuuuuccc gucuguucgu gaucuucucg acaccgcuuc 2040
agcuuuguac cgggaauccu uagaguccuc ugaucauugu ucgccucacc auacagcacu 2100
caggcaagca auccugugcu ggggugaguu gaugacucua gccaccuggg uggguaauaa 2160 2019264591
uuuggaagau ccagcaucca gagauuuggu ggucaauuau guuaauacua auauggguuu 2220
aaaaaucagg caacuauugu gguuucacau uuccugucuu acuuuuggga gagaaaccgu 2280
ucuugaguau uuggugucuu uuggagugug gauucgcacu ccuccugcuu auagaccacc 2340
aaaugccccu auccuaucaa cacuuccgga gacuacuguu guuagacgaa gaggcagguc 2400
cccucgaaga agaacucccu cgccucgcag acgaagaucu caaucgccgc gucgcagaag 2460
aucugcaucu ccagcuuccc aauguuagua uuccuuggac ucacaaggug ggaaacuuua 2520
cggggcugua uucuucuacu auaccugucu uuaauccgga uuggcaaacu ccuucuuuuc 2580
caaauaucca uuugcaucaa gacauuauaa cuaaauguga acaauuugug ggcccucuca 2640
caguaaauga gaaacgaaga uuaaaacuag uuaugccugc cagauuuuuc ccaaacucua 2700
cuaaauauuu accauuagac aaagguauca aaccguauua uccagaaaau guaguuaauc 2760
auuacuucca gaccagacau uauuuacaua cccuuuggaa ggcggguauu cuauauaaga 2820
gagaaacauc ccguagcgcu ucauuuugug ggucaccaua uacuugggaa caagaucuac 2880
agcauggggc uuucuuggac ggucccucuc gaguggggaa agaaccuuuc caccagcaau 2940
ccucuaggau uccuucccga ucaccaguug gacccagcau ucagagcaaa uaccaacaau 3000
ccagauuggg acuucaaucc caaaaaggac ccuuggccag aggccaacaa gguaggaguu 3060
ggagccuaug gacccggguu caccccucca cacggaggcc uuuuggggug gagcccucag 3120
ucucagggca cacuaacaac uuugccagca gauccgccuc cugccuccac caaucgucag 3180
ucagggaggc agccuacucc caucucucca ccacuaagag acagucaucc ucaggcccug 3240
caguggaa 3248
<210> 523 <211> 3248 <212> RNA Page 507
SIRIFD00001WOPCTSEQ 13 Nov 2019
<213> Hepatitis B virus
<400> 523 cucuacagca uuccaccaag cucuacaaaa ucccaaaguc aggggccugu auuuuccugc 60
ugguggcucc aguucaggga uagugaaccc uguuccgacu auugccucuc acaucucguc 120
aaucuucucc aggauugggg acccugcacc gaacauggag aacaucacau caggauuccu 180
aggaccccug cucguguuac aggcgggguu uuucuuguug acaagaaucc ucacaauacc 240 2019264591
acagagucua gacucguggu ggacuucucu caauuuucua gggggagugc ccgugugucc 300
uggccuaaau ucgcaguccc caaccuccaa ucacucacca aucuccuguc cuccaacuug 360
uccuggcuau cgcuggaugu gucugcggcg uuuuaucaua uuccucuuca uccugcugcu 420
augccucauc uucuuguugg uucuucugga cuaucaaggu auguugcccg uuuguccucu 480
gauuccagga uccucgacca ccaguacggg acccugcaaa accugcacga cuccugcuca 540
aggcaacucu auguaucccu cauguugcug uacaaaaccu ucggacggaa auugcaccug 600
uauucccauc ccaucaucuu gggcuuucgc aaaauaccua ugggaguggg ccucaguccg 660
uuucucuugg cucaguuuac uagugccauu uguucagugg uucguagggc uuucccccac 720
ugucuggcuu ucagcuauau ggaugaugug guauuggggg ccaaaucugu acaacaucuu 780
gagucccuuu auaccgcugu uaccaauuuu cuuuugucuu uggguauaca ucuaaacccu 840
accaaaacaa aaagaugggg uuauuccuua aauuuuaugg gauauguaau uggaaguugg 900
gguacuuugc cacaagaaca caucacacag aaaauuaagc aauguuuucg gaaacucccu 960
guuaacaggc caauugauug gaaagucugu caacgaauaa cuggucuguu ggguuucgcu 1020
gcuccuuuua cccaaugugg uuacccugcc uuaaugccuu uauaugcaug uauacaagcu 1080
aagcaggcuu uuacuuucuc gccaacuuau aaggccuuuc ucuguaaaca auacaugaac 1140
cuuuaccccg uugcuaggca acggcccggu cugugccaag uguuugcuga cgcaaccccc 1200
acugguuggg gcuuggccau cggccaucag cgcaugcgug gaaccuuugu ggcuccucug 1260
ccgauccaua cugcggaacu ccuagcugcu uguuuugcuc gcagccgguc uggagcaaaa 1320
cucauuggga cugacaauuc ugucguccuu ucucggaaau auacauccuu uccauggcug 1380
cuaggcugug cugccaacug gauccuucgc gggacguccu uuguuuacgu cccgucagcg 1440
cugaauccag cggacgaccc cucccggggc cguuuggggc ucugucgccc ccuucuccgu 1500 Page 508
SIRIFD00001WOPCTSEQ 13 Nov 2019
cugccguucc ugccgaccac ggggcgcacc ucucuuuacg cggucucccc gucugugccu 1560
ucucaucugc cggaccgugu gcacuucgcu ucaccucugc acguuacaug gaaaccgcca 1620
ugaacaccuc ucaucaucug ccaaggcagu uauauaagag gacucuugga cuguuuguua 1680
ugucaacaac cgggguggag aaauacuuca aggacugugu uuuugcugag ugggaagaau 1740
uaggcaauga guccagguua augaccuuug uauuaggagg cuguaggcau aaauuggucu 1800 2019264591
gcgcaccagc accauguaac uuuuucaccu cugccuaauc aucucuuguu cauguccuac 1860
uguucaagcc uccaagcugu gccuugggug gcuuuagggc auggauagaa caacuuugcc 1920
auauggccuu uuuggcuuag acauugaccc uuauaaagaa uuuggagcua cuguggaguu 1980
gcucucguuu uugccuucug acuuuuuccc gucuguucgu gaucuucucg acaccgcuuc 2040
agcuuuguac cgggaauccu uagaguccuc ugaucauugu ucgccucacc auacagcacu 2100
caggcaagca auccugugcu ggggugaguu gaugacucua gcuaccuggg uggguaauaa 2160
uuuggaagau ccagcaucca gagauuuggu ggucaauuau guuaauacua auauggguuu 2220
aaaaaucagg caacuauugu gguuucacau uuccugucuu acuuuuggga gagaaaccgu 2280
ucuugaguau uuggugucuu uuggagugug gauucgcacu ccuccugcuu auagaccacc 2340
aaaugccccu auccuaucaa cacuuccgga gacuacuguu guuagacgaa gaggcagguc 2400
cccucgaaga agaacucccu cgccucgcag acgaagaucu caaucgccgc gucgcagaag 2460
aucugcaucu ccagcuuccc aauguuagua uuccuuggac ucacaaggug ggaaacuuua 2520
cggggcugua uucuucuacu auaccugucu uuaauccuga uuggcaaacu ccuucuuuuc 2580
caaauaucca uuugcaucaa gacauuauaa cuaaauguga acaauuugug ggcccucuca 2640
caguaaauga gaaacgaaga uuaaaacuag uuaugccugc cagauuuuuc ccaaacucua 2700
cuaaauauuu accauuagac aaagguauca aaccguauua uccagaaaau guaguuaauc 2760
auuacuucca gaccagacau uauuuacaua cccuuuggaa ggcggguauu cuauauaaga 2820
gagaaacauc ccguagcgcu ucauuuugug ggucaccaua uacuugggaa caagaucuac 2880
agcauggggc uuucuuggac ggucccucuc gaguggggaa agaaccuuuc caccagcaau 2940
ccucuaggau uccuucccga ucaccaguug gacccagcau ucagagcaaa uaccaacaau 3000
ccagauuggg acuucaaucc caaaaaggac ccuuggccag aggccaacaa gguaggaguu 3060 Page 509
SIRIFD00001WOPCTSEQ 13 Nov 2019
ggagccuaug gacccggguu caccccucca cacggaggcc uuuuggggug gagcccucag 3120
ucucagggca cacuaacaac uuugccagca gauccgccuc cugccuccac caaucgucag 3180
ucagggaggc agccuacucc caucucucca ccacuaagag acagucaucc ucaggccaug 3240
caguggaa 3248 2019264591
<210> 524 <211> 3215 <212> RNA <213> Hepatitis B virus
<400> 524 gaacucaaca caguuccacc aagcacuguu ggauccgaga gucagggguc uguauuuucc 60
ugcugguggc uccaguucag aaacacagaa cccuguuccg acuauugccu cucucacauc 120
aucaaucuuc ucgaagacug gggacccugc uaugaacaug gagaacauca caucaggacu 180
ccuaggaccc cuucucgugu uacaggcggu guguuucuug uugacaaaaa uccucacaau 240
accacagagu cuagacucgu gguggacuuc ucucaauuuu cuagggguac cacccgggug 300
uccuggccaa aauucgcagu ccccaaucuc caaucacuua ccaaccuccu guccuccaac 360
uuguccuggc uaucguugga ugugucugcg gcguuuuauc aucuuccucu ucauccugcu 420
gcuaugccuc aucuucuugu ugguucuucu ggacuaucaa gguauguugc ccgugugucc 480
ucuacuucca ggaucuacaa ccaccagcac gggacccugc aaaaccugca ccacucuugc 540
ucaaggaacc ucuauguuuc ccuccugcug cuguaccaaa ccuucggacg gaaauugcac 600
cuguauuccc aucccaucau cuugggcuuu cggaaaauac cuaugggagu gggccucagc 660
ccguuucucu uggcucaguu uacuagugca auuuguucag uggugcguag ggcuuucccc 720
cacugucugg cuuuuaguua uauggaugau uugguauugg gggccaaauc ugugcagcau 780
cuugaguccc uuuauaccgc uguuaccaau uuuuuguuau cugugggcau ccauuuaaac 840
acagcuaaaa caaaauggug ggguuauucc uuacacuuua uggguuauau cauugggagu 900
ugggggacau ugccucagga acauauugug caaaaaauca aaaauugcuu ucgcaaacuu 960
cccguuaauc gacccauuga uuggaaaguc ugucaacgaa uugugggucu uuugggcuuu 1020
gcagccccuu uuacucaaug ugguuauccu gcucucaugc ccuuauaugc cuguauuacc 1080
Page 510
SIRIFD00001WOPCTSEQ 13 Nov 2019
gcuaaacagg cuuuuguuuu cucgccaacu uacaaggccu uucucuguaa acaauacaug 1140
aaccuuuacc ccguugcucg gcaacggcca ggccuuugcc aaguguuugc ugacgcaacc 1200
cccacuggcu ggggcuuggc gauuggccau cagcgcaugc gcggaaccuu uguggcuccu 1260
cugccgaucc auacugcgga acuccuagca gcuuguuucg cucgcagccg gucuggagcg 1320
gacauuaucg gcacugacaa cuccguuguc cugucucgga aguacaccuc cuucccaugg 1380 2019264591
cugcuaggcu gugcugccaa cuggauccug cgcgggacgu ccuuuguuua cgucccgucg 1440
gcgcugaauc cugcggacga ccccucucgu ggucgccugg ggcucugccg cccccuucuc 1500
cgccuuccgu uccggccgac gacagggcgc accucucuuu acgcggacuc cccgccugug 1560
ccuuuucauc agccggcccg ugugcacuuc gguucaccuc ugcacgucgc auggagacca 1620
ccgugaacgc cccucaaagc uugccaacac ccuuacauaa aaggacucuu ggacuuucgc 1680
cccggucaac gaccuggauu gaggaauaca ucaaagacug uguguuuaag gacugggagg 1740
agucggggga ggaguugagg uuaaaggucu uuguauuagg aggcuguagg cauaaauugg 1800
ucuguucacc agcaccaugc aacuuuuuca ccucugccua aucaucuuuu guucaugucc 1860
cacuguucaa gccuccaagc ugugccuugg guggcuuugg ggcauggaca uugacccuua 1920
uaaagaauuu ggagcuucug uggaguuacu cucauuuuug ccuucugacu ucuucccguc 1980
uguccgggac cuacucgaca ccgcuucagc ccucuaccga gaugccuuag aaucacccga 2040
acauugcacc cccaaccaua cugcucucag gcaagcuauu cugugcuggg gugaguuaau 2100
gacuuuggcu uccugggugg gcaauaauuu agaggauccu gcggcuagag aucuaguagu 2160
uaauuauguc aacacuaaua ugggccuaaa aauuagacaa uuacuauggu uucauauuuc 2220
cugccuuaca uuuggaagag aaacuguucu ugaguauuug gugucuuuug gaguguggau 2280
ucgcacucca ccugcuuaua gaccaccaaa ugccccuauc cuaucaacac uuccggagac 2340
uacuguuguu agacaacgag gcagggcccc uagaagaaga acucccucgc cucgcagacg 2400
aagaucucaa ucaccgcguc gcagaagauc ucaaucucca gcuucccaau guuaguaucc 2460
cuuggacuca uaagguggga aacuuuaccg gucuuuacuc cucuacugua ccuguuuuca 2520
auccugauug guuaacuccu ucuuuuccug acauucaccu acaucaagau uugauacaaa 2580
aaugugagca auuugugcgc ccacucacua aaaaugaagu gagacgauug aaauuaauua 2640
Page 511
SIRIFD00001WOPCTSEQ 13 Nov 2019
ugccagcaag guuuuauccc aaagcuacua aauacuuccc uuuggauaaa gguauuaaac 2700
cauauuaucc agagaaugug guuaaucauu acuucaaaac uacacacuau uuacauacuu 2760
uguggaaggc aagaauucua uauaagagag aauccacaca uagcgccuca uuuugugggu 2820
caccauauuc cugggaacaa gaacuacagc augggagcac cucucucaac ggcgagaagg 2880
ggcaugggac agaaucucuc ugugcccaau ccucugggau ucuuuccaga ccaccaguug 2940 2019264591
gauccacuau ucagagcaaa uuccagcagu cccgauuggg acuucaacac aaacaaggac 3000
aauuggccaa uggcaaacaa gguaggagug ggaggcuuug guccaggguu cacaccccca 3060
cacgguggcc uucuggggug gagcccucag gcacagggca uucugacaac cucgccacca 3120
gauccaccuc ccgcuuccac caaucggagg ucaggaagga aaccaacccc agucucucca 3180
ccucuaaggg acacacaucc acaggccaug cagug 3215
<210> 525 <211> 3215 <212> RNA <213> Hepatitis B virus
<400> 525 gaacucaaca caguuccacc aagcacuguu ggauccgaga guaagggguc uguaucuucc 60
ugcugguggc uccaguucag aaacacagaa cccuguuccg acuauugccu cucucacauc 120
aucaaucuuc ucgaagacug gggacccugc uaugaacaug gagaacauca caucaggacu 180
ccuaggaccc cuucucgugu uacaggcggu guguuucuug uugacaaaaa uccucacaau 240
accacagagu cuagacucgu gguggacuuc ucucaauuuu cuagggguac cacccgggug 300
uccuggccaa aauucgcagu ccccaaucuc caaucacuua ccaaccuccu guccuccaac 360
uuguccuggc uaucguugga ugugucugcg gcguuuuauc aucuuccucu ucauccugcu 420
gcuaugccuc aucuucuugu ugguucuucu ggacuaucaa gguauguugc ccgugugucc 480
ucuacuucca ggaucuacaa ccaccagcac gggucccugc aaaaccugca ccacucuugc 540
ucaaggaacc ucuauguuuc ccuccugcug cuguaccaaa ccuucggacg gaaauugcac 600
cuguauuccc aucccaucau cuugggcuuu cggaaaauac cuaugggagu gggccucagc 660
ccguuucucu uggcucaguu uacuagugca auuuguucag uggugcguag ggcuuucccc 720
cacugucugg cuuuuaguua uauggaugau uugguauugg gggccaaauc ugugcagcau 780 Page 512
SIRIFD00001WOPCTSEQ 13 Nov 2019
cuugaguccc uuuauaccgc uguuaccaau uuuuuguuau cugugggcau ccauuuaaac 840
acagcuaaaa caaaauggug ggguuauucc uuacacuuua uggguuauau cauugggagu 900
ugggggacau ugccucagga acauauugug caaaaaauca aagauugcuu ucgcaaacuu 960
ccuguuaauc gacccauuga uuggaaaguc ugucaacgaa uugugggucu uuugggcuuu 1020
gcagccccuu uuacucaaug ugguuauccu gcucucaugc ccuuauaugc cuguauuacc 1080 2019264591
gcuaaacagg cuuuuguuuu cucgccaacu uacaaggccu uucucuguaa acaauacaug 1140
aaccuuuacc ccguugcucg gcaacggcca ggccuuugcc aaguguuugc ugacgcaacc 1200
cccacuggcu ggggcuuggc gauuggccau cagcgcaugc gcggaaccuu uguggcuccu 1260
cugccgaucc auacugcgga acuccuagca gcuuguuucg cucgcagccg gucuggagcg 1320
gacauuaucg gcacugacaa cuccguuguc cugucucgga aguacaccuc cuucccaugg 1380
cugcuaggcu gugcugccaa cuggauccug cgcgggacgu ccuuuguuua cgucccgucg 1440
gcgcugaauc cugcggacga ccccucucgu ggucgccugg ggcucugccg cccccuucuc 1500
cgccuuccgu uccggccgac gacgggucgc accucucuuu acgcggacuc cccgccugug 1560
ccuucucauc ugccggcccg ugugcacuuc gcuucaccuc ugcacgucgc auggagacca 1620
ccgugaacgc cccucaaagc uugccaacaa ccuuacauaa gaggacucuu ggacuuucac 1680
cccggucaac gaccuggauu gaggaauaca ucaaagacug uguguuuaag gacugggagg 1740
agucggggga ggaguugagg uuaaaggucu uuguauuagg aggcuguagg cauaaauugg 1800
ucuguucacc agcaccaugc aacuuuuuca ccucugccua aucaucuuuu guucaugucc 1860
cacuguucaa gccuccaagc ugugccuugg guggcuuugg ggcauggaca uugacccuua 1920
uaaagaauuu ggagcuucug uggaguuacu cucauuuuug ccuucugacu ucuucccguc 1980
uguccgggac cuacucgaca ccgcuucagc ccucuaccga gaugccuuag aaucucccga 2040
acauugcacc cccaaccaua cugcucucag gcaagcuauu cugugcuggg gugaguuaau 2100
gacuuuggcu uccugggugg gcaauaauuu agaggauccu gcggcuagag aucuaguagu 2160
uaauuauguc aacacuaaua ugggccuaaa aauuagacaa uuacuauggu uucauauuuc 2220
cugccuuaca uuuggaagag auacuguucu ugaguauuug gugucuuuug gaguguggau 2280
ucgcacucca ccugcuuaua gaccaccaaa ugccccuauc cuaucaacac uuccggagac 2340 Page 513
SIRIFD00001WOPCTSEQ 13 Nov 2019
uacuguuguu agacaacgag gcagggcccc uagaagaaga acucccucgc cucgcagacg 2400
aagaucucaa ucaccgcguc gccgaagauc ucaaucucca gcuucccaau guuaguaucc 2460
cuuggacuca uaagguggga aacuuuaccg gucuuuacuc cucuacugua ccuguuuuca 2520
auccugauug guuaacuccu ucuuuuccug acauucaccu acaucaagau uugauacaaa 2580
aaugugaaca auuuguaggc ccacucacua aaaaugaagu gagacgauug aaauuaauua 2640 2019264591
ugccagcaag auuuuauccc aaaguuacua aauacuuccc uuuggauaaa gguauuaaac 2700
cauauuaucc agagcaugug guuaaucauu acuucaaaac uagacacuau uuacauacuu 2760
uguggaaggc aggaauucua uauaagagag aauccacaca uagcgccuca uuuugugggu 2820
caccauauuc cugggaacaa gagcuacagc augggagcac cucucucaac ggcgagaagg 2880
ggcaugggac agaaucuuuc ugugcccaau ccucugggau ucuuuccaga ccaccaguug 2940
gauccacuau ucagagcaaa uuccagcagu cccgauuggg acuucaacac aaacaaggac 3000
aauuggccaa uggcaaacaa gguaggagug ggaggcuuug guccaggguu cacaccccca 3060
cacgguggcc uucuggggug gagcccucag gcacagggca uucugacaac cucgccaccc 3120
gauccaccuc ccgcuuccac caaucggagg ucaggaagga aaccaacccc agucucucca 3180
ccucuaaggg acacacaucc acaggccaug cagug 3215
<210> 526 <211> 3215 <212> RNA <213> Hepatitis B virus
<400> 526 gaacucaaca caguuccacc aagcacuguu ggauccgaga guaagggguc uguauuuucc 60
ugcugguggc uccaguucag aaacacagaa cccugcuccg acuauugccu cucucacauc 120
aucaaucuuc ucgaagacug gggacccugc uaugaacaug gagaacauca caucaggacu 180
ccuaagaccc cuucucgugu uacaggcggu guguuucuug uugacaaaaa uccucacaau 240
accacagagu cuagacucgu gguggacuuc ucucaauuuu cuagggguac cacccgggug 300
uccuggccaa aauucgcagu ccccaaucuc caaucacuua ccaaccuccu guccuccaac 360
uuguccuggc uaucguugga ugugucugcg gcguuuuauc aucuuccucu ucauccugcu 420
Page 514
SIRIFD00001WOPCTSEQ 13 Nov 2019
gcuaugccuc aucuucuugu ugguucuucu ggacuaucaa gguauguugc ccgugugucc 480
ucuacuucca ggaucuacaa ccaccagcac gggacccugc aaaaccugca ccacucuugc 540
ucaaggaacc ucuauguuuc ccucuugcug cuguaccaaa ccuucggacg gaaauugcac 600
cuguauuccc aucccaucau cuugggcuuu cggaaaauac cuaugggagu gggccucagc 660
ccguuucucu uggcucaguu uacuagugca auuuguucag uggugcguag ggcuuucccc 720 2019264591
cacugucugg cuuuuaguua uauggaugau uugguauugg gggccaaauc ugugcagcau 780
cuugaguccc uuuauaccgc uguuaccaau uuuuuguuau cugugggcau ccauuugaac 840
acagcuaaaa caaaauggug ggguuauucc uuacacuuua uggguuauau aauugggagu 900
ugggggaccu ugccucagga acauauugug cauaaaauca aagauugcuu ucgcaaacuu 960
cccgugaaua gacccauuga uuggaagguu ugucaacgca uugugggucu uuugggcuuu 1020
gcagccccuu uuacucaaug ugguuauccu gcucucaugc ccuuguaugc cuguauuacc 1080
gcuaagcagg cuuuuguuuu cucgccaacu uacaaggccu uucucuguaa acaauacaug 1140
aaccuuuacc ccguugcucg gcaacggcca ggccuuugcc aaguguuugc ugacgcaacc 1200
cccacuggcu ggggcuuggc gauuggccau cagcgcaugc gcggaaccuu uguggcuccu 1260
cugccgaucc auacugcgga acuccuagca gcuuguuucg cucgcagcag gucuggagcg 1320
gacauuaucg gcacugacaa cucuguuguc cuuucucgga aguacaccuc cuucccaugg 1380
cugcucggcu gugcugccaa cuggauccug cgcgggacgu ccuuugucua cgucccgucg 1440
gcgcugaauc cugcggacga ccccucucgu ggucgcuugg ggcucugccg cccucuucuc 1500
cgccugccgu uccggccgac gacgggucgc accucucuuu acgcggacuc cccgccugug 1560
ccuucucauc ugccggcccg ugugcacuuc gcuucaccuc ugcacgucgc auggagacca 1620
ccgugaacgc cccuuggaac uugccaacaa ccuugcauaa gaggacucuu ggucuuucgc 1680
cccggucaac gaccuggauu gaggaauaca ucaaagacug uguauuuaag gacugggagg 1740
agucggggga ggaguugagg uuaaaggucu uuguauuagg aggcuguagg cauaaauugg 1800
ucuguucacc agcaccaugc aacuuuuuca ccucugccua aucaucuuuu guucaugucc 1860
cacuguucaa gccuccaagc ugugccuugg guggcuuugg ggcauggaca uugacccuua 1920
uaaagaauuu ggagcuucug uggaguuacu cucauuuuug ccuucugacu ucuucccguc 1980
Page 515
SIRIFD00001WOPCTSEQ 13 Nov 2019
uguccgggac cugcucgaca ccgcuucagc ccucuaccga gaugccuuag aaucacccga 2040
acauugcacc cccaaccaca cugcucucag gcaagcuauu uugugcuggg gugaguugau 2100
gaccuuggcu uccugggugg gcaauaauuu agaggauccu gcagcaagag aucuaguagu 2160
uaauuauguc aauacuaaca ugggccuaaa aauuagacaa uuauuauggu uucacauuuc 2220
cugccuuaca uuuggaagag aaacugugcu ugaguauuug gugucuuuug gaguguggau 2280 2019264591
ucgcacucca ccugcuuaua gaccaccaaa ugccccuauc cuaucaacac uuccggagac 2340
uacuguuguu agacaacgag gcagggcccc uagaagaaga acucccucgc cucgcagacg 2400
aagaucucaa ucaccgcguc gcagaagauc ucaaucucca gcuucccaau guuaguauuc 2460
cuuggacuca uaagguggga aacuuuaccg gucuuuacuc cucuacuaua ccuguuuuua 2520
auccugacug gcuaacuccc ucuuuuccug acauucacuu gcaucaagau cugauacaaa 2580
aaugugaaca auuuguaggc ccacucacua caaaugaaag gagacgauug aaacuaauua 2640
ugccagcuag guuuuauccc aaaguuacua aauacuuccc uuuggauaaa gguauuaagc 2700
cuuacuaucc agagaaugug guuaaucauu acuucaaaac uagacauuau uuacauacuu 2760
uguggaaggc aggaauucua uauaagagag aauccacaca uagcgccuca uuuugugggu 2820
caccauauuc cugggaacaa gagcuacagc augggagcac cucucucaac ggcgagaagg 2880
ggcaugggac agaaccuuuc ugugcccaau ccucugggau ucuuuccaga ccaccaguug 2940
gauccacuau ucagagcaaa uuccagcagu cccgauuggg acuucaacac aaacaaggac 3000
aauuggccaa uggcaaacaa gguaggagug ggaggcuucg guccaggguu cacaccccca 3060
cacgguggcc uucuggggug gagcccucag gcacagggca uucugacaac cucgccacca 3120
gauccaccuc cugcuuccac caaucggagg ucaggaagaa agccaacccc agucucucca 3180
ccucuaaggg acacacaucc acaggccaug cagug 3215
Page 516
Claims (23)
1. A double-stranded short interfering nucleic acid (siNA) molecule comprising a sense strand comprising the nucleotide sequence 5'-B gCCgaUCCaUaCUgCggaaUsU-3'(SEQ ID NO:326), and an antisense strand comprising the nucleotide sequence 5' UUCCgCagUaUggaUCggCUsU-3'(SEQ ID NO:225); wherein B is an inverted abasic moiety; A, C, G, and U are ribose A, C, G, and U, respectively; bolded nucleotides A, C, G, and U are 2' -methyl (2'-OMe) A, 2'-OMe C, 2'-OMe G, and 2'-OMe U, respectively; a, c, g, and u are 2' fluoro A, 2'-fluoro C, 2'-fluoro G, and 2'-fluoro U, respectively; italicized nucleotides A, C, G, and U are deoxy A, deoxy C, deoxy G, and deoxy U, respectively; and s is a phosphorothioate linkage.
2. A composition comprising the double-stranded short interfering nucleic acid (siNA) according to claim 1 and a pharmaceutically acceptable carrier or diluent.
3. A composition comprising: (a) the double-stranded short interfering nucleic acid (siNA) of claim 1; (b) a cationic lipid; (c) cholesterol; (d) DSPC; and (e) PEG-DMG.
4. A composition comprising: (a) the double-stranded short interfering nucleic acid (siNA) of claim 1; (b) (13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien-l-amine; (c) cholesterol; (d) DSPC; and (e) PEG-DMG.
5. The composition according to claim 4, wherein the (13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien--amine, cholesterol, DSPC, and PEG-DMG have a molar ratio of 50:30:10:2 respectively.
6. A composition comprising the composition according to any one of claims 3-5 and a pharmaceutically acceptable carrier or diluent.
7. A kit comprising the double-stranded short interfering nucleic acid (siNA) according to claim 1, or the composition according to any one of claims 2-6.
8. A method of treating a human subject suffering from a condition which is mediated by the action, or by loss of action, of Hepatitis B Virus (HBV), comprising administering to said subject an effective amount of the double-stranded short interfering nucleic acid (siNA) molecule of claim 1, or the composition of any one of claims 2-6, or the kit of claim 7, thereby treating the subject.
9. The method according to claim 8, wherein the condition is HBV infection.
10. The method according to claim 8, wherein the condition is hepatocellular carcinoma.
11. The method according to claim 8, wherein the condition is liver disease.
12. The method according to claim 11, wherein the liver disease is cirrhosis.
13. A method of treating a human subject suffering from Hepatitis B Virus (HBV) infection, comprising administering to said subject an effective amount of the double-stranded short interfering nucleic acid (siNA) molecule of claim 1, or the composition of any one of claims 2-6, or the kit of claim 7, wherein the progression of HBV to hepatocellular carcinoma is reduced in the subject, thereby treating the subject.
14. A method of treating a human subject suffering from hepatocellular carcinoma, comprising administering to said subject an effective amount of the double-stranded short interfering nucleic acid (siNA) molecule of claim 1, or the composition of any one of claims 2-6, or the kit of claim 7, and an anti-cancer agent, thereby treating the subject.
15. The method of claim 14, wherein the anti-cancer agent is sorafenib.
16. Use of a double-stranded short interfering nucleic acid (siNA) molecule of claim 1 or the composition of any one of claims 2-6 in the manufacture of a medicament for treating a human subject suffering from a condition which is mediated by the action, or by loss of action, of Hepatitis B Virus (HBV).
17. The use according to claim 16, wherein the condition is HBV infection.
18. The use according to claim 16, wherein the condition is hepatocellular carcinoma.
19. The use according to claim 16, wherein the condition is liver disease.
20. The use according to claim 19, wherein the liver disease is cirrhosis.
21. Use of a double-stranded short interfering nucleic acid (siNA) molecule of claim 1 or the composition of any one of claims 2-6 in the manufacture of a medicament for treating a human subject suffering from Hepatitis B Virus (HBV) infection to reduce the progression of HBV to hepatocellular carcinoma.
22. Use of a double-stranded short interfering nucleic acid (siNA) molecule of claim 1 or the composition of any one of claims 2-6 and an anti-cancer agent in the manufacture of a medicament for treating a human subject suffering from hepatocellular carcinoma.
23. The use of claim 22, wherein said anti-cancer agent is sorafenib.
Sirna Therapeutics, Inc. Patent Attorneys for the Applicant/Nominated Person SPRUSON & FERGUSON
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| Application Number | Priority Date | Filing Date | Title |
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| AU2019264591A AU2019264591B2 (en) | 2010-08-17 | 2019-11-13 | RNA interference mediated inhibition of hepatitis B virus (HBV) gene expression using short interfering nucleic acid (siNA) |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US37455510P | 2010-08-17 | 2010-08-17 | |
| US61/374,555 | 2010-08-17 | ||
| AU2011292261A AU2011292261B2 (en) | 2010-08-17 | 2011-08-12 | RNA interference mediated inhibition of Hepatitis B virus (HBV) gene expression using short interfering nucleic acid (siNA) |
| PCT/US2011/047512 WO2012024170A2 (en) | 2010-08-17 | 2011-08-12 | RNA INTERFERENCE MEDIATED INHIBITION OF HEPATITIS B VIRUS (HBV) GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA) |
| AU2015210336A AU2015210336B2 (en) | 2010-08-17 | 2015-08-04 | RNA interference mediated inhibition of hepatitis B virus (HBV) gene expression using short interfering nucleic acid (siNA) |
| AU2017216560A AU2017216560A1 (en) | 2010-08-17 | 2017-08-18 | RNA interference mediated inhibition of hepatitis B virus (HBV) gene expression using short interfering nucleic acid (siNA) |
| AU2019264591A AU2019264591B2 (en) | 2010-08-17 | 2019-11-13 | RNA interference mediated inhibition of hepatitis B virus (HBV) gene expression using short interfering nucleic acid (siNA) |
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| AU2017216560A Abandoned AU2017216560A1 (en) | 2010-08-17 | 2017-08-18 | RNA interference mediated inhibition of hepatitis B virus (HBV) gene expression using short interfering nucleic acid (siNA) |
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| AU2017216560A Abandoned AU2017216560A1 (en) | 2010-08-17 | 2017-08-18 | RNA interference mediated inhibition of hepatitis B virus (HBV) gene expression using short interfering nucleic acid (siNA) |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998028004A1 (en) * | 1996-12-24 | 1998-07-02 | The Crown In The Right Of The Queensland Department Of Health (Sir Albert Sakzewski Virus Research Centre) | Hepatitis delta particle containing a fusion protein immunogen |
| US20100145038A1 (en) * | 2003-11-24 | 2010-06-10 | Merck & Co., Inc. | RNA INTERFERENCE MEDIATED INHIBITION OF GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA) |
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| CN101603042B (en) * | 2008-06-13 | 2013-05-01 | 厦门大学 | RNA interference target for treating hepatitis B virus infection |
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- 2017-08-18 AU AU2017216560A patent/AU2017216560A1/en not_active Abandoned
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998028004A1 (en) * | 1996-12-24 | 1998-07-02 | The Crown In The Right Of The Queensland Department Of Health (Sir Albert Sakzewski Virus Research Centre) | Hepatitis delta particle containing a fusion protein immunogen |
| US20100145038A1 (en) * | 2003-11-24 | 2010-06-10 | Merck & Co., Inc. | RNA INTERFERENCE MEDIATED INHIBITION OF GENE EXPRESSION USING SHORT INTERFERING NUCLEIC ACID (siNA) |
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| AU2015210336B2 (en) | 2017-05-25 |
| AU2015210336A1 (en) | 2015-08-27 |
| AU2019264591A1 (en) | 2019-12-05 |
| AU2017216560A1 (en) | 2017-09-07 |
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