US20200392510A1

US20200392510A1 - Modulators of dnm2 expression

Info

Publication number: US20200392510A1
Application number: US16/962,136
Authority: US
Inventors: Susan M. Freier; Huynh-Hoa Bui; Susan F. Murray; Brett P. Monia; Shuling Guo
Original assignee: Ionis Pharmaceuticals Inc
Current assignee: Ionis Pharmaceuticals Inc
Priority date: 2018-01-15
Filing date: 2019-01-15
Publication date: 2020-12-17
Also published as: AU2019206731A2; IL276007A; CL2020001851A1; AU2019206731A1; WO2019140452A1; US20190241896A1; CO2020008769A2; BR112020014425A2; MX2020007369A; PE20211242A1; TW201934129A; SG11202006711UA; CN111902537A; US10865414B2; EP3740575A1; RU2020127049A; KR20200113214A; CA3088522A1; JP2021511072A; MA51645A

Abstract

The present embodiments provide methods, compounds, and compositions useful for inhibiting DNM2 expression, which may be useful for treating, preventing, or ameliorating a disease associated with DNM2.

Description

SEQUENCE LISTING

The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled BIOL0322WOSEQ_ST25.txt created Jan. 10, 2019 which is 784 kb in size. The information in the electronic format of the sequence listing is incorporated herein by reference in its entirety.

FIELD

The present embodiments provide methods, compounds, and compositions useful for inhibiting DNM2 expression, which can be useful for treating, preventing, or ameliorating a disease associated with DNM2.

BACKGROUND

Dynamin 2 (DNM2) is a large GTPase that is involved in membrane trafficking and microtubule dynamics (See, e.g., Durieux et al. J Mol Med, 88, 339 (2010)). Mutations in DNM2 are associated with several diseases, including centronuclear myopathy (CNM) and Charcot-Marie-Tooth disease. DNM2 is also associated with Duchenne's Muscular Dystrophy (See, e.g., WO 2015/055859 and WO 2016/170162). CNM and CNM-like myopathy are also associated with mutations in MTM1, BIN1, RYR1, TTN, CCDC78, and MTMR14

SUMMARY

Certain embodiments provided herein are directed to potent and tolerable compounds and compositions useful for inhibiting DNM2 expression, which can be useful for treating, preventing, ameliorating, or slowing progression of diseases, such as centronuclear myopathy (CNM), Charcot-Marie-Tooth disease (CMT), and Duchenne's Muscular Dystrophy (DMD). Certain embodiments provided herein comprise modified oligonucleotides complementary to a DNM2 nucleic acid that potently reduce DNM2 expression in animals.
One object of the present invention is a compound comprising a modified oligonucleotide 8 to 50 linked nucleosides in length having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134. Another object of the present invention is a compound comprising a modified oligonucleotide 9 to 50 linked nucleosides in length having a nucleobase sequence comprising at least 9 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134. Another object of the present invention is a compound comprising a modified oligonucleotide 10 to 50 linked nucleosides in length having a nucleobase sequence comprising at least 10 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134. Another object of the present invention is a compound comprising a modified oligonucleotide 11 to 50 linked nucleosides in length having a nucleobase sequence comprising at least 11 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134. Another object of the present invention is a compound comprising a modified oligonucleotide 12 to 50 linked nucleosides in length having a nucleobase sequence comprising at least 12, at least 13, at least 14, or at least 15 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134. Another object of the present invention is a compound comprising a modified oligonucleotide 16 to 50 linked nucleosides in length having a nucleobase sequence comprising the nucleobase sequence of any of SEQ ID NOs: 7-3134. Another object of the present invention is a compound comprising a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 7-3134. Another object of the present invention is a compound comprising a modified oligonucleotide 8 to 50 linked nucleosides in length complementary within nucleobases 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1, wherein said modified oligonucleotide is at least 85%, 90%, 95%, or 100% complementary to SEQ ID NO: 1. Another object of the present invention is a compound comprising a modified oligonucleotide 8 to 50 linked nucleosides in length having a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1, wherein the nucleobase sequence of the modified oligonucleotide is at least 85%, 90%, 95%, or 100% complementary to SEQ ID NO: 1. Another object of the present invention is a compound comprising a modified oligonucleotide 8 to 50 linked nucleosides in length complementary within nucleobases 83,573-87,287 or 87,359-90,915 of SEQ ID NO: 1, wherein said modified oligonucleotide is at least 85%, 90%, 95%, or 100% complementary to SEQ ID NO: 1. Another object of the present invention is a compound comprising a modified oligonucleotide complementary to intron 1 of a human DNM2 pre-mRNA. Another object of the present invention is a compound comprising a modified oligonucleotide complementary to intron 11 of a human DNM2 pre-mRNA. Another object of the present invention is a compound comprising a modified oligonucleotide complementary to intron 12 of a human DNM2 pre-mRNA. Another object of the present invention is a compound comprising a modified oligonucleotide complementary to intron 13 of a human DNM2 pre-mRNA. Another object of the present invention is a compound comprising a modified oligonucleotide complementary to intron 14 of a human DNM2 pre-mRNA. Another object of the present invention is a compound comprising a modified oligonucleotide complementary to exon 10 of a human DNM2 pre-mRNA or human DNM2 mRNA. Another object of the present invention is a compound comprising a modified oligonucleotide complementary to the 3′-UTR of a human DNM2 pre-mRNA or human DNM2 mRNA. Another object of the present invention is a compound comprising a modified oligonucleotide 16 to 50 linked nucleosides in length having a nucleobase sequence comprising any of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. Another object of the present invention is a compound comprising a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. Another object of the present invention is a compound comprising a modified oligonucleotide having a nucleobase sequence comprising SEQ ID NO: 2879.
Another object of the present invention is a compound comprising a modified oligonucleotide 16 to 50 linked nucleosides in length having a nucleobase sequence comprising any of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232, wherein the modified oligonucleotide comprises:

- a gap segment consisting of 8-12 linked 2′-deoxynucleosides;
- a 5′ wing segment consisting of 1-7 linked nucleosides; and
- a 3′ wing segment consisting of 1-7 linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each terminal wing nucleoside comprises a modified sugar.
Another object of the present invention is a compound comprising a modified oligonucleotide 16 linked nucleosides in length having a nucleobase sequence consisting of the sequence recited in SEQ ID NO: 2879, 3056, 2123, 2189, 2453, 2160, or 2232, wherein the modified oligonucleotide comprises

- a gap segment consisting of 10 linked 2′-deoxynucleosides;
- a 5′ wing segment consisting of 3 linked nucleosides; and
- a 3′ wing segment consisting of 3 linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein each nucleoside of each wing segment comprises a cEt sugar moiety; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine.
In one embodiment, the oligonucleotide is at least 80%, 85%, 90%, 95% or 100% complementary to any of SEQ ID NO: 1, 2, 3, or 3135. In one embodiment, the modified oligonucleotide comprises at least one modified internucleoside linkage. In one embodiment, the at least one modified internucleoside linkage is a phosphorothioate internucleoside linkage. In one embodiment, the modified oligonucleotide comprises at least one bicyclic sugar. In one embodiment, the at least one bicyclic sugar is selected from the group consisting of LNA, ENA, and cEt. In one embodiment, the at least one bicyclic sugar moiety is a cEt sugar moiety.
In one embodiment, the modified oligonucleotide comprises at least one 5-methylcytosine. In one embodiment, the modified oligonucleotide comprises:
a gap segment consisting of linked 2′-deoxynucleosides;
a 5′ wing segment consisting of linked nucleosides; and
a 3′ wing segment consisting of linked nucleosides;
wherein the gap segment is positioned immediately adjacent to and between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar moiety.
In one embodiment, the compound is single-stranded. In one embodiment, the compound is double-stranded.
In one embodiment, the compound comprises at least one unmodified ribosyl sugar moiety. In one embodiment, the compound comprises at least one unmodified 2′-deoxyribosyl sugar moiety.
In one embodiment, the modified oligonucleotide consists of 10 to 30 linked nucleosides. In one embodiment, the modified oligonucleotide consists of 12 to 30 linked nucleosides. In one embodiment, the modified oligonucleotide consists of 15 to 30 linked nucleosides. In one embodiment, the modified oligonucleotide consists of 16 to 20 linked nucleosides.
Another object of the present invention is a compound comprising a modified oligonucleotide according to the following formula: Gks Tks Tks Tds Ads Tds Tds Ads Tds Ads Gds Gds Gds mCks Tks Tk; wherein,

- A=an adenine,
- mC=a 5-methylcytosine
- G=a guanine,
- T=a thymine,
- k=a cEt sugar moiety,
- d=a 2′-deoxyribosyl sugar moiety, and
- s=a phosphorothioate internucleoside linkage.

In one embodiment, the compound of the invention comprises a conjugate group. In one embodiment, the compound of the invention consists of the modified oligonucleotide and the conjugate group.
Another object of the present invention is a compound according to the following formula:
In one embodiment, the compound consists of the modified oligonucleotide.
The present invention further relates to a compound consisting of a pharmaceutically acceptable salt form of any one of the compounds described herein. In one embodiment, the pharmaceutically acceptable salt is a sodium salt. In one embodiment, the pharmaceutically acceptable salt is a potassium salt.
The present invention also relates to a pharmaceutical composition comprising a compound as described herein and at least one pharmaceutically acceptable carrier or diluent.
The present invention further relates to a chirally enriched population of the compounds described herein, wherein the population is enriched for modified oligonucleotides comprising at least one particular phosphorothioate internucleoside linkage having a particular stereochemical configuration. In one embodiment, the population is enriched for modified oligonucleotides comprising at least one particular phosphorothioate internucleoside linkage having the (Sp) configuration. In one embodiment, the population is enriched for modified oligonucleotides comprising at least one particular phosphorothioate internucleoside linkage having the (Rp) configuration. In one embodiment, the population is enriched for modified oligonucleotides having a particular, independently selected stereochemical configuration at each phosphorothioate internucleoside linkage. In one embodiment, the population is enriched for modified oligonucleotides having the (Sp) configuration at each phosphorothioate internucleoside linkage. In one embodiment, the population is enriched for modified oligonucleotides having the (Rp) configuration at each phosphorothioate internucleoside linkage. In one embodiment, the population is enriched for modified oligonucleotides having the (Rp) configuration at one particular phosphorothioate internucleoside linkage and the (Sp) configuration at each of the remaining phosphorothioate internucleoside linkages. In one embodiment, the population is enriched for modified oligonucleotides having at least 3 contiguous phosphorothioate internucleoside linkages in the Sp, Sp, and Rp configurations, in the 5′ to 3′ direction. In one embodiment, all of the phosphorothioate internucleoside linkages of the modified oligonucleotide are stereorandom.
The present invention further relates to a pharmaceutical composition comprising the population of compounds described herein and at least one pharmaceutically acceptable diluent or carrier.
The present invention further relates to a compound described herein, a pharmaceutical composition comprising said compound and at least one pharmaceutically acceptable carrier or diluent, or a pharmaceutical composition comprising the population of compounds described herein and at least one pharmaceutically acceptable carrier or diluent, for use in therapy.
The present invention further relates to a compound described herein, a pharmaceutical composition comprising said compound and at least one pharmaceutically acceptable carrier or diluent, or a pharmaceutical composition comprising the population of compounds described herein and at least one pharmaceutically acceptable carrier or diluent, for use in treating, preventing, or ameliorating centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease.
The present invention also relates to a method of treating, preventing, or ameliorating a disease associated with DNM2 in an individual comprising administering to the individual a compound or composition described herein, thereby treating, preventing, or ameliorating the disease.
The present invention also relates to a method of treating, preventing, or ameliorating a disease associated with DNM2 in an individual comprising administering to the individual a compound comprising a modified oligonucleotide 100% complementary to an exon 10, an intron, or the 3′-UTR of a DNM2 nucleic acid transcript, thereby treating, preventing, or ameliorating the disease.
In one embodiment, the disease is centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease. In one embodiment, the compound is single-stranded. In one embodiment, the DNM2 nucleic acid transcript is a pre-mRNA. In one embodiment, the disease is X-linked myotubular myopathy, autosomal recessive centronuclear myopathy, or autosomal dominant centronuclear myopathy. In one embodiment, the individual has at least one mutation in at least one gene selected from among MTM1, BIN1, and DNM2. In one embodiment, the administering increases body weight or muscle strength.
The present invention further relates to a method of inhibiting expression of DNM2 in a cell comprising contacting the cell with a single-stranded compound comprising a modified oligonucleotide 100% complementary to exon 10, an intron, or the 3′-UTR of a DNM2 nucleic acid transcript, thereby inhibiting expression of DNM2 in the cell. In one embodiment, the method is an in vitro method. In one embodiment, the cell is in the muscle of an individual. In one embodiment, the individual has, or is at risk of having, a centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease.
The present invention further relates to a method of increasing body weight or muscle strength in an individual having, or at risk of having, a disease associated with DNM2 comprising administering a single-stranded compound comprising a modified oligonucleotide 100% complementary to a DNM2 nucleic acid transcript to the individual, thereby increasing body weight or muscle strength in the individual. In one embodiment, the individual has, or is at risk of having, centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease.
In one embodiment, the compound is a compound as described herein. In one embodiment, the compound is a member of the chirally enriched population described herein. In one embodiment, the compound is a component of the pharmaceutical composition described herein.
In one embodiment, the compound is administered to the individual via subcutaneous injection. In one embodiment, the compound is administered to the individual via intramuscular injection. In one embodiment, the compound is administered to the individual via intravenous injection.
The present invention further relates to the use of a single-stranded compound comprising a modified oligonucleotide 100% complementary to exon 10, intron 1, intron 11, intron, 12, intron 13, intron 14, or the 3′-UTR of a DNM2 nucleic acid transcript for treating, preventing, or ameliorating a disease associated with DNM2. In one embodiment, the disease is centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease.
The present invention further relates to the use of the compound described herein or the composition described herein for treating, preventing, or ameliorating a disease associated with DNM2.
The present invention further relates to the use of the compound described herein or the composition described herein in the manufacture of a medicament for treating, preventing, or ameliorating a disease associated with DNM2.
In one embodiment, the disease is centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease. In one embodiment, the disease is X-linked myotubular myopathy, autosomal recessive centronuclear myopathy, or autosomal dominant centronuclear myopathy. In one embodiment, the disease is associated with a mutation in at least one gene selected from among MTM1, BIN1, and DNM2. The present invention further relates to the use of the compound described herein or the composition described herein in the preparation of a medicament for treating, preventing, or ameliorating a disease associated with DNM2. In one embodiment, the disease is centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease. In one embodiment, the disease is X-linked myotubular myopathy, autosomal recessive centronuclear myopathy, or autosomal dominant centronuclear myopathy. In one embodiment, the the disease is associated with a mutation in at least one gene selected from among MTM1, BIN1, and DNM2.

DETAILED DESCRIPTION

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the embodiments, as claimed. Herein, the use of the singular includes the plural unless specifically stated otherwise. As used herein, the use of “or” means “and/or” unless stated otherwise. Furthermore, the use of the term “including” as well as other forms, such as “includes” and “included”, is not limiting.
The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. All documents, or portions of documents, cited in this application, including, but not limited to, patents, patent applications, articles, books, treatises, and GenBank and NCBI reference sequence records are hereby expressly incorporated by reference for the portions of the document discussed herein, as well as in their entirety.
It is understood that the sequence set forth in each SEQ ID NO contained herein is independent of any modification to a sugar moiety, an internucleoside linkage, or a nucleobase. As such, compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an internucleoside linkage, or a nucleobase.
As used herein, “2′-deoxynucleoside” means a nucleoside comprising 2′-H(H) ribosyl sugar moiety, as found in naturally occurring deoxyribonucleic acids (DNA). In certain embodiments, a 2′-deoxynucleoside may comprise a modified nucleobase or may comprise an RNA nucleobase (uracil).
As used herein, “2′-substituted nucleoside” or “2-modified nucleoside” means a nucleoside comprising a 2′-substituted or 2′-modified sugar moiety. As used herein, “2′-substituted” or “2-modified” in reference to a furanosyl sugar moiety means a sugar moiety comprising at least one 2′-substituent group other than H or OH.
As used herein, “administration” or “administering” refers to routes of introducing a compound or composition provided herein to an individual to perform its intended function. An example of a route of administration that can be used includes, but is not limited to, administration by inhalation.
As used herein, “administered concomitantly” or “co-administration” means administration of two or more compounds in any manner in which the pharmacological effects of both are manifest in the patient. Concomitant administration does not require that both compounds be administered in a single pharmaceutical composition, in the same dosage form, by the same route of administration, or at the same time. The effects of both compounds need not manifest themselves at the same time. The effects need only be overlapping for a period of time and need not be coextensive. Concomitant administration or co-administration encompasses administration in parallel or sequentially.
As used herein, “animal” refers to a human or non-human animal, including, but not limited to, mice, rats, rabbits, dogs, cats, pigs, and non-human primates, including, but not limited to, monkeys and chimpanzees.
As used herein, “antisense activity” means any detectable and/or measurable change attributable to the hybridization of an antisense compound to its target nucleic acid. In certain embodiments, antisense activity is a decrease in the amount or expression of a target nucleic acid or protein encoded by such target nucleic acid compared to target nucleic acid levels or target protein levels in the absence of the antisense compound.
As used herein, “antisense compound” means a compound comprising an antisense oligonucleotide and optionally one or more additional features, such as a conjugate group or terminal group.
As used herein, “antisense oligonucleotide” means an oligonucleotide having a nucleobase sequence that is at least partially complementary to a target nucleic acid.
As used herein, “ameliorate” in reference to a treatment means improvement in at least one symptom relative to the same symptom in the absence of the treatment. In certain embodiments, amelioration is the reduction in the severity or frequency of a symptom or the delayed onset or slowing of progression in the severity or frequency of a symptom.
As used herein, “bicyclic nucleoside” or “BNA” means a nucleoside comprising a bicyclic sugar moiety. As used herein, “bicyclic sugar” or “bicyclic sugar moiety” means a modified sugar moiety comprising two rings, wherein the second ring is formed via a bridge connecting two of the atoms in the first ring thereby forming a bicyclic structure. In certain embodiments, the first ring of the bicyclic sugar moiety is a furanosyl moiety. In certain embodiments, the bicyclic sugar moiety does not comprise a furanosyl moiety.
As used herein, “cEt” or “constrained ethyl” means a bicyclic sugar moiety, wherein the first ring of the bicyclic sugar moiety is a ribosyl sugar moiety, the second ring of the bicyclic sugar is formed via a bridge connecting the 4′-carbon and the 2′-carbon, the bridge has the formula 4′-CH(CH₃)—O-2′, and the methyl group of the bridge is in the S configuration. A cEt bicyclic sugar moiety is in the β-D configuration.
As used herein, “chirally enriched population” means a plurality of molecules of identical molecular formula, wherein the number or percentage of molecules within the population that contain a particular stereochemical configuration at a particular chiral center is greater than the number or percentage of molecules expected to contain the same particular stereochemical configuration at the same particular chiral center within the population if the particular chiral center were stereorandom. Chirally enriched populations of molecules having multiple chiral centers within each molecule may contain one or more sterorandom chiral centers. In certain embodiments, the molecules are modified oligonucleotides. In certain embodiments, the molecules are compounds comprising modified oligonucleotides.
As used herein, “complementary” in reference to an oligonucleotide means that at least 70% of the nucleobases of such oligonucleotide or one or more regions thereof and the nucleobases of another nucleic acid or one or more regions thereof are capable of hydrogen bonding with one another when the nucleobase sequence of the oligonucleotide and the other nucleic acid are aligned in opposing directions. Complementary nucleobases are nucleobase pairs that are capable of forming hydrogen bonds with one another. Complementary nucleobase pairs include adenine (A) and thymine (T), adenine (A) and uracil (U), cytosine (C) and guanine (G), 5-methyl cytosine (^mC) and guanine (G). Complementary oligonucleotides and/or nucleic acids need not have nucleobase complementarity at each nucleoside. Rather, some mismatches are tolerated. As used herein, “fully complementary” or “100% complementary” in reference to oligonucleotides means that such oligonucleotides are complementary to another oligonucleotide or nucleic acid at each nucleoside of the oligonucleotide.
As used herein, “conjugate group” means a group of atoms that is directly or indirectly attached to an oligonucleotide. Conjugate groups include a conjugate moiety and a conjugate linker that attaches the conjugate moiety to the oligonucleotide.
As used herein, “conjugate linker” means a group of atoms comprising at least one bond that connects a conjugate moiety to an oligonucleotide.
As used herein, “conjugate moiety” means a group of atoms that is attached to an oligonucleotide via a conjugate linker.
As used herein, “contiguous” in the context of an oligonucleotide refers to nucleosides, nucleobases, sugar moieties, or internucleoside linkages that are immediately adjacent to each other. For example, “contiguous nucleobases” means nucleobases that are immediately adjacent to each other in a sequence.
As used herein, “double-stranded antisense compound” means an antisense compound comprising two oligomeric compounds that are complementary to each other and form a duplex, and wherein one of the two said oligomeric compounds comprises an antisense oligonucleotide.
As used herein, “effective amount” means the amount of compound sufficient to effectuate a desired physiological outcome in an individual in need of the compound. The effective amount may vary among individuals depending on the health and physical condition of the individual to be treated, the taxonomic group of the individuals to be treated, the formulation of the composition, assessment of the individual's medical condition, and other relevant factors.
As used herein, “efficacy” means the ability to produce a desired effect.
As used herein “DNM2” means any Dynamin 2 nucleic acid or protein. “DNM2 nucleic acid” means any nucleic acid encoding DNM2. For example, in certain embodiments, a DNM2 nucleic acid includes a DNA chromosomal region encoding DNM2, an RNA transcribed from DNA encoding DNM2 (e.g., a pre-mRNA transcript), and an mRNA transcript encoding DNM2.
As used herein, “expression” includes all the functions by which a gene's coded information is converted into structures present and operating in a cell. Such structures include, but are not limited to, the products of transcription and translation. Methods for measuring expression levels (such as a transcription level or a translation level) are well-known in the field and include, but are not limited to, RT-PCR, RT-qPCR, Northern Blot, hybridization techniques such as, for example, use of microarrays, and combinations thereof including but not limited to, hybridization of amplicons obtained by RT-PCR, sequencing such as, for example, next-generation DNA sequencing (NGS) or RNA-seq (also known as “Whole Transcriptome Shotgun Sequencing”) and the like, immunohistochemistry, Multiplex methods (Luminex), western blot, enzyme-linked immunosorbent assay (ELISA), sandwich ELISA, multiplex ELISA, electrochemiluminescence (ECL) (Elecsys®, Roche Diagnostics), enzyme-linked fluorescent assay (ELFA) (such as VIDAS®, Biomdrieux), fluorescent-linked immunosorbent assay (FLISA), enzyme immunoassay (EIA), radioimmunoassay (RIA), flow cytometry (FACS), surface plasmon resonance (SPR), biolayer interferometry (BLI), immunochromatographic assay (ICA) (such as NEXUS IB10, Sphingotech) and mass spectrometry-based approaches.
As used herein, “gapmer” means an oligonucleotide, such as an antisense oligonucleotide, comprising an internal segment having a plurality of nucleosides that support RNase H cleavage positioned between external segments, each having one or more nucleosides, wherein the nucleosides comprising the internal segment are chemically distinct from the immediately adjacent nucleoside or nucleosides comprising the external segments. The internal segment may be referred to as the “gap” or “gap segment” and the external segments may be referred to as the “wings” or “wing segments”.
As used herein, “hybridization” means the pairing or annealing of complementary oligonucleotides and/or nucleic acids. While not limited to a particular mechanism, the most common mechanism of hybridization involves hydrogen bonding, which may be, for example, Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding, between complementary nucleobases.
As used herein, “individual” means a human or non-human animal selected for treatment or therapy.
As used herein, “inhibiting the expression or activity” refers to a reduction or blockade of the expression or activity relative to the expression or activity in an untreated or control sample and does not necessarily indicate a total elimination of expression or activity. In one embodiment, inhibiting the expression or activity refers to a reduction or blockade of the expression or activity relative to the expression or activity in an untreated or control sample of at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%. In one embodiment, inhibiting the expression or activity refers to a reduction or blockade of the expression or activity relative to the expression or activity in an untreated or control sample of at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%.
As used herein, the term “about” preceding a figure means plus or less 10% of the value of said figure.
As used herein, the terms “internucleoside linkage” means a group or bond that forms a covalent linkage between adjacent nucleosides in an oligonucleotide. As used herein “modified internucleoside linkage” means any internucleoside linkage other than a naturally occurring, phosphate internucleoside linkage. Non-phosphate linkages are referred to herein as modified internucleoside linkages. “Phosphorothioate linkage” means a modified phosphate linkage in which one of the non-bridging oxygen atoms is replaced with a sulfur atom. A phosphorothioate internucleoside linkage is a modified internucleoside linkage. Modified internucleoside linkages include linkages that comprise abasic nucleosides. As used herein, “abasic nucleoside” means a sugar moiety in an oligonucleotide or oligomeric compound that is not directly connected to a nucleobase. In certain embodiments, an abasic nucleoside is adjacent to one or two nucleosides in an oligonucleotide.
As used herein, “linker-nucleoside” means a nucleoside that links, either directly or indirectly, an oligonucleotide to a conjugate moiety. Linker-nucleosides are located within the conjugate linker of an oligomeric compound. Linker-nucleosides are not considered part of the oligonucleotide portion of an oligomeric compound even if they are contiguous with the oligonucleotide.
As used herein, “non-bicyclic modified sugar” or “non-bicyclic modified sugar moiety” means a modified sugar moiety that comprises a modification, such as a substituent, that does not form a bridge between two atoms of the sugar to form a second ring.
As used herein, “linked nucleosides” are nucleosides that are connected in a continuous sequence (i.e. no additional nucleosides are present between those that are linked).
As used herein, “mismatch” or “non-complementary” means a nucleobase of a first oligonucleotide that is not complementary with the corresponding nucleobase of a second oligonucleotide or target nucleic acid when the first and second oligomeric compound are aligned.
As used herein, “modulating” refers to changing or adjusting a feature in a cell, tissue, organ or organism. For example, modulating DNM2 expression can mean to increase or decrease the level of an DNM2 RNA and/or a DNM2 protein in a cell, tissue, organ or organism. A “modulator” effects the change in the cell, tissue, organ or organism. For example, a compound that modulates DNM2 expression can be a modulator that decreases the amount of a DNM2 RNA and/or a DNM2 protein in a cell, tissue, organ or organism.
As used herein, “MOE” means methoxyethyl. “2′-MOE” or “2′-O-methoxyethyl” means a 2′-OCH₂CH₂OCH₃group in place of the 2′OH group of a ribosyl ring.
As used herein, “motif” means the pattern of unmodified and/or modified sugar moieties, nucleobases, and/or internucleoside linkages, in an oligonucleotide.
As used herein, “naturally occurring” means found in nature.
As used herein, “nucleobase” means an unmodified nucleobase or a modified nucleobase. As used herein an “unmodified nucleobase” is adenine (A), thymine (T), cytosine (C), uracil (U), and guanine (G). As used herein, a modified nucleobase is a group of atoms capable of pairing with at least one unmodified nucleobase. A universal base is a nucleobase that can pair with any one of the five unmodified nucleobases.
As used herein, “nucleobase sequence” means the order of contiguous nucleobases in a nucleic acid or oligonucleotide independent of any sugar or internucleoside linkage modification.
As used herein, “nucleoside” means a moiety comprising a nucleobase and a sugar moiety. The nucleobase and sugar moiety are each, independently, unmodified or modified. As used herein, “modified nucleoside” means a nucleoside comprising a modified nucleobase and/or a modified sugar moiety.
As used herein, “oligomeric compound” means a compound consisting of an oligonucleotide and optionally one or more additional features, such as a conjugate group or terminal group.
As used herein, “oligonucleotide” means a strand of linked nucleosides connected via internucleoside linkages, wherein each nucleoside and internucleoside linkage may be modified or unmodified. Unless otherwise indicated, oligonucleotides consist of 8-50 linked nucleosides. As used herein, “modified oligonucleotide” means an oligonucleotide, wherein at least one nucleoside or internucleoside linkage is modified. As used herein, “unmodified oligonucleotide” means an oligonucleotide that does not comprise any nucleoside modifications or internucleoside modifications.
As used herein, “pharmaceutically acceptable carrier or diluent” means any substance suitable for use in administering to an animal. Certain such carriers enable pharmaceutical compositions to be formulated as, for example, liquids, powders, or suspensions that can be aerosolized or otherwise dispersed for inhalation by a subject. In certain embodiments, a pharmaceutically acceptable carrier or diluent is sterile water; sterile saline; or sterile buffer solution. In one embodiment, the pharmaceutically acceptable carrier or diluent does not produce an adverse, allergic or other untoward reaction when administered to an animal, preferably a human. It includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. For human administration, preparations should meet sterility, pyrogenicity, general safety and purity standards as required by regulatory offices, such as, for example, FDA Office or EMA.
As used herein “pharmaceutically acceptable salts” means physiologically and pharmaceutically acceptable salts of compounds, such as oligomeric compounds, i.e., salts that retain the desired biological activity of the parent compound and do not impart undesired toxicological effects thereto.
As used herein “pharmaceutical composition” means a mixture of substances suitable for administering to a subject. For example, a pharmaceutical composition may comprise an antisense compound and an aqueous solution.
As used herein, “phosphorus moiety” means a group of atoms comprising a phosphorus atom. In certain embodiments, a phosphorus moiety comprises a mono-, di-, or tri-phosphate, or phosphorothioate.
As used herein “prodrug” means a therapeutic agent in a form outside the body that is converted to a differentform within the body or cells thereof. Typically conversion of a prodrug within the body is facilitated by the action of an enzymes (e.g., endogenous or viral enzyme) or chemicals present in cells or tissues and/or by physiologic conditions.
As used herein, “RNAi compound” means an antisense compound that acts, at least in part, through RISC or Ago2 to modulate a target nucleic acid and/or protein encoded by a target nucleic acid. RNAi compounds include, but are not limited to double-stranded siRNA, single-stranded RNA (ssRNA), and microRNA, including microRNA mimics. In certain embodiments, an RNAi compound modulates the amount, activity, and/or splicing of a target nucleic acid. The term RNAi compound excludes antisense oligonucleotides that act through RNase H.
As used herein, the term “single-stranded” in reference to an antisense compound means such a compound consisting of one oligomeric compound that is not paired with a second oligomeric compound to form a duplex. “Self-complementary” in reference to an oligonucleotide means an oligonucleotide that at least partially hybridizes to itself. A compound consisting of one oligomeric compound, wherein the oligonucleotide of the oligomeric compound is self-complementary, is a single-stranded compound. A single-stranded antisense or oligomeric compound may be capable of binding to a complementary oligomeric compound to form a duplex, in which case the compound would no longer be single-stranded.
As used herein, “standard cell assay” means any of the assays described in Examples 1-9, and reasonable variations thereof.
As used herein, “standard in vivo experiment” means the procedure described in Example 10 and reasonable variations thereof.
As used herein, “stereorandom chiral center” in the context of a population of molecules of identical molecular formula means a chiral center having a random stereochemical configuration. For example, in a population of molecules comprising a stereorandom chiral center, the number of molecules having the (S) configuration of the stereorandom chiral center may be but is not necessarily the same as the number of molecules having the (R) configuration of the stereorandom chiral center. The stereochemical configuration of a chiral center is considered random when it is the result of a synthetic method that is not designed to control the stereochemical configuration. In certain embodiments, a stereorandom chiral center is a stereorandom phosphorothioate internucleoside linkage.
As used herein, “sugar moiety” means an unmodified sugar moiety or a modified sugar moiety. As used herein, “unmodified sugar moiety” means a 2′-OH(H) ribosyl moiety, as found in RNA (an “unmodified RNA sugar moiety”), or a 2′-H(H) moiety, as found in DNA (an “unmodified DNA sugar moiety”). As used herein, “modified sugar moiety” or “modified sugar” means a modified furanosyl sugar moiety or a sugar surrogate. As used herein, modified furanosyl sugar moiety means a furanosyl sugar comprising a non-hydrogen substituent in place of at least one hydrogen of an unmodified sugar moiety. In certain embodiments, a modified furanosyl sugar moiety is a 2′-substituted sugar moiety. Such modified furanosyl sugar moieties include bicyclic sugars and non-bicyclic sugars. As used herein, “sugar surrogate” means a modified sugar moiety having other than a furanosyl moiety that can link a nucleobase to another group, such as an internucleoside linkage, conjugate group, or terminal group in an oligonucleotide. Modified nucleosides comprising sugar surrogates can be incorporated into one or more positions within an oligonucleotide and such oligonucleotides are capable of hybridizing to complementary oligomeric compounds or nucleic acids.
As used herein, “target nucleic acid,” “target RNA,” “target RNA transcript” and “nucleic acid target” mean a nucleic acid that an antisense compound is designed to affect.
As used herein, “target region” means a portion of a target nucleic acid to which an antisense compound is designed to hybridize.
As used herein, “terminal group” means a chemical group or group of atoms that is covalently linked to a terminus of an oligonucleotide.
As used herein, “terminal wing nucleoside” means a nucleoside that is located at the terminus of a wing segment of a gapmer. Any wing segment that comprises or consists of at least two nucleosides has two termini: one that is immediately adjacent to the gap segment; and one that is at the end opposite the gap segment. Thus, any wing segment that comprises or consists of at least two nucleosides has two terminal nucleosides, one at each terminus.
As used herein, “therapeutically effective amount” means an amount of a compound, pharmaceutical agent, or composition that provides a therapeutic benefit to an individual. In certain embodiments, the therapeutically effective amount refers to the level or amount of a compound that treats or ameliorates symptoms of a disease associated with DNM2. In certain embodiments, the therapeutically effective amount may be administered prior to the onset of the disease associated with DNM2 or may be administered after initiation of the disease associated with DNM2.
As used herein, “treat” refers to administering a compound or pharmaceutical composition to an animal in order to effect an alteration or improvement of a disease, disorder, or condition in the animal. In one embodiment, a subject is successfully treated if, after receiving a therapeutically effective amount of a compound according to the present invention, the subject shows observable and/or measurable reduction or relief to some extent of one or more of the symptoms associated with the specific disease associated with DNM2, such as but not limited to reduced morbidity and mortality, improvement in quality of life issues, increased body weight, preservation or increase in muscle strength, decrease in time taken to rise from the floor, decrease in nine-meter walking time, decrease in time taken to climb four stairs, increased ability to lift weight, increased distance of a 6 minute walk, increased leg function grade, improved pulmonary function and/or cardiac function. Parameters for assessing successful treatment and improvement in the disease are readily measurable by routine procedures familiar to a physician.

Certain Embodiments

Certain embodiments provide methods, compounds and compositions for inhibiting DNM2 expression.
Certain embodiments provide methods, compounds and compositions for inhibiting DNM2 expression as compared to an untreated or control sample. In one embodiment, said inhibition is an inhibition of DNM2 expression of at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%. In one embodiment, inhibiting the expression or activity refers to a reduction or blockade of the expression or activity relative to the expression or activity in an untreated or control sample of at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% as compared to an untreated or control sample.
Certain embodiments provide compounds comprising or consisting of oligonucleotides complementary to a DNM2 nucleic acid. In certain embodiments, the DNM2 nucleic acid has the sequence set forth in RefSeq or GenBank Accession No. NC_000019.10 truncated from nucleosides 10715001 to Ser. No. 10/835,000 (disclosed herein as SEQ ID NO: 1), NM_004945.3 (disclosed herein as SEQ ID NO: 2), NM_001005361.2 (disclosed herein as SEQ ID NO: 3), or NM_001005360.2 (disclosed herein as SEQ ID NO: 3135). In certain embodiments, the compound is an antisense compound or oligomeric compound. In certain embodiments, the compound is single-stranded. In certain embodiments, the compound is double-stranded.
Certain embodiments provide a compound comprising a modified oligonucleotide 8 to 50 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134 or of any sequence having at least about 70% identity with any of the nucleobase sequences of SEQ ID NOs: 7-3134, such as, for example, at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity or more with any of the nucleobase sequences of SEQ ID NOs: 7-3134.
As used herein, the term “identity” refers to the subunit sequence identity between two nucleic acid molecules, such as, two DNA molecules or two RNA molecules. When a subunit position in both of the two molecules is occupied by the same monomeric subunit; e.g., if a position in each of two DNA molecules is occupied by adenine, then they are identical (or homologous) at that position. The homology between two sequences is a direct function of the number of matching or homologous positions; e.g., if half (e.g., five positions in a polymer ten subunits in length) of the positions in two sequences are homologous, the two sequences are 50% homologous; if 90% of the positions (e.g., 9 of 10), are matched or homologous, the two sequences are 90% homologous. Thus, the term “homologous” or “identical”, when used in a relationship between the sequences of two or more nucleic acid molecules, refers to the degree of sequence relatedness between nucleic acid molecules, as determined by the number of matches between strings of two or more nucleotide residues. “Identity” measures the percent of identical matches between the smaller of two or more sequences with gap alignments (if any) addressed by a particular mathematical model or computer program (i.e., “algorithms”). Identity of related polypeptides can be readily calculated by known methods. Such methods include, but are not limited to, those described in Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part 1, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M. Stockton Press, New York, 1991; and Carillo et al., SIAM J. Applied Math. 48, 1073 (1988). Preferred methods for determining identity are designed to give the largest match between the sequences tested. Methods of determining identity are described in publicly available computer programs. Preferred computer program methods for determining identity between two sequences include the GCG program package, including GAP (Devereux et al., Nucl. Acid. Res. 2, 387 (1984); Genetics Computer Group, University of Wisconsin, Madison, Wis.), BLASTP, BLASTN, and FASTA (Altschul et al., J. MoI. Biol. 215, 403-410 (1990)). The BLASTX program is publicly available from the National Center for Biotechnology Information (NCBI) and other sources (BLAST Manual, Altschul et al. NCB/NLM/NIH Bethesda, Md. 20894; Altschul et al., supra). The well-known Smith Waterman algorithm may also be used to determine identity.
In certain embodiments, the compound is an antisense compound or oligomeric compound. In certain embodiments, the compound is single-stranded. In certain embodiments, the compound is double-stranded. In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length.
Certain embodiments provide a compound comprising a modified oligonucleotide 8 to 50 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232 or at least 8 contiguous nucleobases of a sequence having at least 70% identity with any of the nucleobase sequences of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In certain embodiments, the nucleobase sequence of the modified oligonucleotide comprises or consists of SEQ ID NO: 2879 or a sequence having at least 70% identity with SEQ ID NO: 2879. In certain embodiments, the nucleobase sequence of the modified oligonucleotide comprises or consists of SEQ ID NO: 3056 or a sequence having at least 70% identity with SEQ ID NO: 3056. In certain embodiments, the nucleobase sequence of the modified oligonucleotide comprises or consists of SEQ ID NO: 2123 or a sequence having at least 70% identity with SEQ ID NO: 2123. In certain embodiments, the nucleobase sequence of the modified oligonucleotide comprises or consists of SEQ ID NO: 2189 or a sequence having at least 70% identity with SEQ ID NO: 2189. In certain embodiments, the nucleobase sequence of the modified oligonucleotide comprises or consists of SEQ ID NO: 2453 or a sequence having at least 70% identity with SEQ ID NO: 2453. In certain embodiments, the nucleobase sequence of the modified oligonucleotide comprises or consists of SEQ ID NO: 2160 or a sequence having at least 70% identity with SEQ ID NO: 2160. In certain embodiments, the nucleobase sequence of the modified oligonucleotide comprises or consists of SEQ ID NO: 2232 or a sequence having at least 70% identity with SEQ ID NO: 2232. In certain embodiments, the compound is an antisense compound or oligomeric compound. In certain embodiments, the compound is single-stranded. In certain embodiments, the compound is double-stranded. In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide has a nucleobase sequence comprising at least 12 contiguous nucleobases of any of SEQ ID Numbers from 7 to 3134 or of any sequence having at least about 70% identity with any of the nucleobase sequences of SEQ ID NOs: 7-3134, such as, for example, at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity or more with any of the nucleobase sequences of SEQ ID NOs: 7-3134.
Certain embodiments provide a compound comprising a modified oligonucleotide 10 to 50 linked nucleosides in length and having a nucleobase sequence comprising at least 10 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134 or of any sequence having at least about 70% identity with any of the nucleobase sequences of SEQ ID NOs: 7-3134, such as, for example, at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity or more with any of the nucleobase sequences of SEQ ID NOs: 7-3134. In certain embodiments, the compound is an antisense compound or oligomeric compound. In certain embodiments, the compound is single-stranded. In certain embodiments, the compound is double-stranded. In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length.
Certain embodiments provide a compound comprising a modified oligonucleotide 11 to 50 linked nucleosides in length and having a nucleobase sequence comprising at least 11 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134 or of any sequence having at least about 70% identity with any of the nucleobase sequences of SEQ ID NOs: 7-3134, such as, for example, at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity or more with any of the nucleobase sequences of SEQ ID NOs: 7-3134. In certain embodiments, the compound is an antisense compound or oligomeric compound. In certain embodiments, the compound is single-stranded. In certain embodiments, the compound is double-stranded. In certain embodiments, the modified oligonucleotide is 11 to 30 linked nucleosides in length.
Certain embodiments provide a compound comprising a modified oligonucleotide 12 to 50 linked nucleosides in length and having a nucleobase sequence comprising at least 12 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134 or of any sequence having at least about 70% identity with any of the nucleobase sequences of SEQ ID NOs: 7-3134, such as, for example, at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity or more with any of the nucleobase sequences of SEQ ID NOs: 7-3134. In certain embodiments, the compound is an antisense compound or oligomeric compound. In certain embodiments, the compound is single-stranded. In certain embodiments, the compound is double-stranded. In certain embodiments, the modified oligonucleotide is 12 to 30 linked nucleosides in length.
In certain embodiments, the compound comprises a modified oligonucleotide 16 linked nucleosides in length. In certain embodiments, the compound is an antisense compound or oligomeric compound.
Certain embodiments provide a compound comprising a modified oligonucleotide 16 to 50 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 7-3134 or of any sequence having at least about 70% identity with any of the nucleobase sequences of SEQ ID NOs: 7-3134, such as, for example, at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity or more with any of the nucleobase sequences of SEQ ID NOs: 7-3134. In certain embodiments, the compound is an antisense compound or oligomeric compound. In certain embodiments, the compound is single-stranded. In certain embodiments, the compound is double-stranded. In certain embodiments, the modified oligonucleotide is 16 to 30 linked nucleosides in length.
Certain embodiments provide a compound comprising a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 7-3134 or of any sequence having at least about 70% identity with any of the nucleobase sequences of SEQ ID NOs: 7-3134, such as, for example, at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity or more with any of the nucleobase sequences of SEQ ID NOs: 7-3134. In certain embodiments, the compound is an antisense compound or oligomeric compound. In certain embodiments, the compound is single-stranded. In certain embodiments, the compound is double-stranded.
In certain embodiments, compounds comprise or consist of modified oligonucleotides complementary to an intron of a DNM2 nucleic acid transcript. In certain embodiments, modified oligonucleotides are complementary to intron 1, intron 2, intron 3, intron 4, intron 5, intron 6, intron 7, intron 8, intron 9, intron 9-10a, intron 10, intron 10a-11, intron 11, intron 12, intron 13, intron 14, intron 15, intron 16, intron 17, intron 18, intron 19, or intron 19-20a of a DNM2 nucleic acid transcript. In certain such embodiments, modified oligonucleotides are complementary to a sequence within nucleotides 3404-44,737; 44,812-57,478; 57,629-60,702; 60,907-62,117; 62,217-67,959; 68,121-71,563; 71,707-78,719; 78,856-80,371; 80,440-81,060; 80,440-82,379; 81,200-83,485; 82,519-83,485; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 105,090-108,787; 108,990-110,056; 110,222-114,035; 114,269-115,126; 115,379-115,977; or 115,379-115,980 of SEQ ID NO: 1. In certain embodiments, compounds comprise or consist of oligonucleotides having at least an 8, 9, 10, 11, 12, 13, 14, 15, or 16 contiguous nucleobase portion complementary to an equal length portion of intron 1, intron 2, intron 3, intron 4, intron 5, intron 6, intron 7, intron 8, intron 9, intron 9-10a, intron 10, intron 10a-11, intron 11, intron 12, intron 13, intron 14, intron 15, intron 16, intron 17, intron 18, intron 19, or intron 19-20a of a DNM2 nucleic acid transcript. In certain embodiments, such oligonucleotides have at least an 8, 9, 10, 11, 12, 13, 14, 15, or 16 contiguous nucleobase portion complementary to an equal length portion within nucleotides 3404-44,737; 44,812-57,478; 57,629-60,702; 60,907-62,117; 62,217-67,959; 68,121-71,563; 71,707-78,719; 78,856-80,371; 80,440-81,060; 80,440-82,379; 81,200-83,485; 82,519-83,485; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 105,090-108,787; 108,990-110,056; 110,222-114,035; 114,269-115,126; 115,379-115,977; or 115,379-115,980 of SEQ ID NO: 1. In certain embodiments, these compounds are antisense compounds or oligomeric compounds. Compounds comprising a modified oligonucleotide complementary to certain introns of a DNM2 nucleic acid transcript, e.g., intron 1, intron 11, intron 12, intron 13, or intron 14 of a DNM2 pre-mRNA, are generally especially potent and tolerable. Thus, such certain introns can be considered hot spot regions for targeting a DNM2 nucleic acid transcript.
In certain embodiments, compounds comprise or consist of modified oligonucleotides complementary to intron 1, intron 11, intron 12, intron 13, intron 14, exon 10, or the 3′-UTR of a DNM2 nucleic acid transcript. In certain embodiments, modified oligonucleotides are complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1. In certain embodiments, compounds comprise or consist of oligonucleotides having at least an 8, 9, 10, 11, 12, 13, 14, 15, or 16 contiguous nucleobase portion complementary to an equal length portion of intron 1, intron 11, intron 12, intron 13, intron 14, exon 10, or the 3′-UTR of a DNM2 nucleic acid transcript.
In certain embodiments, such oligonucleotides have at least an 8, 9, 10, 11, 12, 13, 14, 15, or 16 contiguous nucleobase portion complementary to an equal length portion within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1. In certain embodiments, these compounds are antisense compounds or oligomeric compounds.
In certain embodiments, a compound comprises a modified oligonucleotide 8 to 50 linked nucleosides in length and having at least an 8, 9, 10, 11, 12, 13, 14, 15, or 16 contiguous nucleobase portion complementary to an equal length portion within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1. In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length.
In certain embodiments, compounds comprise or consist of modified oligonucleotides complementary to intron 1, intron 11, intron 12, intron 13, or intron 14 of a DNM2 nucleic acid transcript. In certain embodiments, modified oligonucleotides are complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; or 97,378-104,979 of SEQ ID NO: 1. In certain embodiments, compounds comprise or consist of oligonucleotides having at least an 8, 9, 10, 11, 12, 13, 14, 15, or 16 contiguous nucleobase portion complementary to an equal length portion of intron 1, intron 11, intron 12, intron 13, or intron 14 of a DNM2 nucleic acid transcript. In certain embodiments, such oligonucleotides have at least an 8, 9, 10, 11, 12, 13, 14, 15, or 16 contiguous nucleobase portion complementary to an equal length portion within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; or 97,378-104,979 of SEQ ID NO: 1. In certain embodiments, these compounds are antisense compounds or oligomeric compounds.
In certain embodiments, a compound comprises a modified oligonucleotide 8 to 50 linked nucleosides in length and having at least an 8, 9, 10, 11, 12, 13, 14, 15, or 16 contiguous nucleobase portion complementary to an equal length portion within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; or 97,378-104,979; of SEQ ID NO: 1. In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length.
In certain embodiments, a compound comprises a modified oligonucleotide 8 to 50 linked nucleosides in length and complementary within nucleotides 89,722-89,737; 83,880-83,895; 90,081-90,096; 94,450-94,465; 11,960-11,975; 93,322-93,337; or 97,855-97,870 of SEQ ID NO: 1. In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length.
In certain embodiments, a compound comprises a modified oligonucleotide 8 to 50 linked nucleosides in length and having a nucleobase sequence comprising at least an 8, 9, 10, 11, 12, 13, 14, 15, or 16 contiguous nucleobase portion of the nucleobase sequence of any one of compound numbers 951799, 949935, 950023, 950089, 951372, 950060, or 950132 (SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232). In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length. In certain embodiments, the nucleobase sequence of the modified oligonucleotide comprises or consists of SEQ ID NO: 2879 or of a sequence having at least 70% identity with SEQ ID NO: 2879. In certain embodiments, the nucleobase sequence of the modified oligonucleotide comprises or consists of SEQ ID NO: 3056 or of a sequence having at least 70% identity with SEQ ID NO: 3056. In certain embodiments, the nucleobase sequence of the modified oligonucleotide comprises or consists of SEQ ID NO: 2123 or of a sequence having at least 70% identity with SEQ ID NO: 2123. In certain embodiments, the nucleobase sequence of the modified oligonucleotide comprises or consists of SEQ ID NO: 2189 or of a sequence having at least 70% identity with SEQ ID NO: 2189. In certain embodiments, the nucleobase sequence of the modified oligonucleotide comprises or consists of SEQ ID NO: 2453 or of a sequence having at least 70% identity with SEQ ID NO: 2453. In certain embodiments, the nucleobase sequence of the modified oligonucleotide comprises or consists of SEQ ID NO: 2160 or of a sequence having at least 70% identity with SEQ ID NO: 2160. In certain embodiments, the nucleobase sequence of the modified oligonucleotide comprises or consists of SEQ ID NO: 2232 or of a sequence having at least 70% identity with SEQ ID NO: 2232.
In certain embodiments, a compound comprises or consists of a modified oligonucleotide 8 to 50 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of compound numbers 951799, 949935, 950023, 950089, 951372, 950060, or 950132 (SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232). In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length.
In certain embodiments, a compound comprises or consists of a modified oligonucleotide having a nucleobase sequence consisting of the nucleobase sequence of any one of compound numbers 951799, 949935, 950023, 950089, 951372, 950060, or 950132 (SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232). In certain embodiments, a compound comprises or consists of a modified oligonucleotide 8 to 50 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of the compound number 951799 (SEQ ID NO: 2879). In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length.
In certain embodiments, a compound comprising or consisting of a modified oligonucleotide complementary to DNM2 is compound number 951799. Out of the over 3,000 compounds that were screened as described in the Examples section below, compound numbers 951799, 949935, 950023, 950089, 951372, 950060, and 950132 emerged as the top lead compounds. In particular, compound number 951799 exhibited the best combination of properties in terms of potency and tolerability out of over 3,000 compounds.
In certain embodiments, any of the foregoing oligonucleotides is a modified oligonucleotide comprising at least one modified internucleoside linkage, at least one modified sugar, and/or at least one modified nucleobase.
In certain embodiments, any of the foregoing modified oligonucleotides comprises at least one modified sugar. In certain embodiments, at least one modified sugar is a bicyclic sugar, such as a cEt bicyclic sugar, an LNA bicyclic sugar, or an ENA bicyclic sugar.
In certain embodiments, the modified oligonucleotide comprises at least one modified internucleoside linkage, such as a phosphorothioate internucleoside linkage.
In certain embodiments, any of the foregoing modified oligonucleotides comprises at least one modified nucleobase, such as 5-methylcytosine.
In certain embodiments, any of the foregoing modified oligonucleotides comprises:

- a gap segment consisting of linked 2′-deoxynucleosides;
- a 5′ wing segment consisting of linked nucleosides; and
- a 3′ wing segment consisting of linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar. In certain embodiments, the modified oligonucleotide is 16 to 50 linked nucleosides in length having a nucleobase sequence comprising the sequence recited in anyone of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232 or a sequence having at least 70% identity with any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In certain embodiments, the modified oligonucleotide is 16 to 50 linked nucleosides in length having a nucleobase sequence comprising the sequence recited in SEQ ID NO: 2879 or a sequence having at least 70% identity with SEQ ID NO: 2879. In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length having a nucleobase sequence comprising the sequence recited in any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232 or a sequence having at least 70% identity with any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In certain embodiments, the modified oligonucleotide is 10 to 30 linked nucleosides in length having a nucleobase sequence comprising the sequence recited in SEQ ID NO: 2879 or a sequence having at least 70% identity with SEQ ID NO: 2879. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length having a nucleobase sequence consisting of the sequence recited in any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232 or a sequence having at least 70% identity with any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length having a nucleobase sequence consisting of the sequence recited in SEQ ID NO: 2879 or a sequence having at least 70% identity with SEQ ID NO: 2879.
In certain embodiments, a compound comprises or consists of a modified oligonucleotide 20-50 linked nucleobases in length having a nucleobase sequence comprising the sequence recited in any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232 or a sequence having at least 70% identity with any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232, wherein the modified oligonucleotide comprises
a gap segment consisting often linked 2′-deoxynucleosides;
a 5′ wing segment consisting of three linked nucleosides; and
a 3′ wing segment consisting of three linked nucleosides;
wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein the nucleosides of the 5′ wing segment each comprise a cEt bicyclic sugar; wherein the nucleosides of the 3′ wing segment each comprises a cEt bicyclic sugar; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide is 16-80 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16-30 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is 16 linked nucleosides in length.
In certain embodiments, a compound comprises or consists of a modified oligonucleotide according to one of the following formulas:

(SEQ ID NO: 2879)

Gks Tks Tks Tds Ads Tds Tds Ads Tds Ads Gds Gds

Gds mCks Tks Tk;

(SEQ ID NO: 3056)

Gks Gks Aks Tds Tds Tds Tds Ads Gds Gds Ads Gds

Gds Tks Gks Ak;

(SEQ ID NO: 2123)

Gks mCks Aks Tds Ads Gds Ads mCds Ads Ads Ads Tds

mCds mCks mCks Ak;

(SEQ ID NO: 2189)

Gks mCks Aks Ads Ads Tds Ads Tds Gds Ads Tds Tds

mCds Aks Tks mCk;

(SEQ ID NO: 2453)

Gks Gks Tks mCds Ads Tds Tds Ads Ads Ads Gds Ads

Tds Tks mCks Tk;

(SEQ ID NO: 2160)

Aks Tks Gks Tds Ads Tds Tds Ads mCds mCds Tds Ads

mCds Gks Gks mCk;

or

(SEQ ID NO: 2232)

Gks Tks Aks mCds Ads Ads Tds Gds Tds Ads Ads Gds

mCds mCks Tks Tk

wherein A=an adenine, mC=a 5-methylcytosine, G=a guanine, T=a thymine, k=a cEt sugar moiety, d=a 2′-deoxyribosyl sugar moiety, and s=a phosphorothioate internucleoside linkage.

In certain embodiments, a compound comprises or consists of compound 951799, or salt thereof, a modified oligonucleotide having the following chemical structure:
In certain embodiments, a compound comprises or consists of the sodium salt of compound 951799 having the following chemical structure:
In any of the foregoing embodiments, the compound or oligonucleotide can be at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% complementary to a nucleic acid encoding DNM2.
In any of the foregoing embodiments, the compound can be single-stranded. In certain embodiments, the compound comprises 2′-deoxyribonucleosides. In certain embodiments, the compound is double-stranded. In certain embodiments, the compound is double-stranded and comprises ribonucleosides. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound.
In any of the foregoing embodiments, the compound can be 8 to 80, 10 to 30, 12 to 50, 13 to 30, 13 to 50, 14 to 30, 14 to 50, 15 to 30, 15 to 50, 16 to 30, 16 to 50, 17 to 30, 17 to 50, 18 to 22, 18 to 24, 18 to 30, 18 to 50, 19 to 22, 19 to 30, 19 to 50, or 20 to 30 linked nucleosides in length. In certain embodiments, the compound comprises or consists of an oligonucleotide.
The present invention thus relates to a modified oligonucleotide or compounds comprising modified oligonucleotides complementary to a DNM2 nucleic acid as described hereinabove.
In certain embodiments, a compound comprises a modified oligonucleotide described herein and a conjugate group. In certain embodiments, the conjugate group is linked to the modified oligonucleotide at the 5′ end of the modified oligonucleotide. In certain embodiments, the conjugate group is linked to the modified oligonucleotide at the 3′ end of the modified oligonucleotide.
In certain embodiments, compounds or compositions provided herein comprise a salt of the modified oligonucleotide. In certain embodiments, the salt is a sodium salt. In certain embodiments, the salt is a potassium salt.
In certain embodiments, the compounds or compositions as described herein are active by virtue of having at least one of an in vitro IC₅₀of less than 300 nM, 200 nM, 100 nM, 80 nM, 50 nM, or 30 nM in a standard cell assay. Standard cell assays are described in the Examples.
In certain embodiments, the compounds or compositions as described herein are highly tolerable as demonstrated by having at least one of an increase an alanine transaminase (ALT) or aspartate transaminase (AST) value of no more than 4 fold, 3 fold, 2 fold, or 1.5 fold over saline treated animals or an increase in liver, spleen, or kidney weight of no more than 30%, 20%, 15%, 12%, 10%, 5%, or 2% compared to control treated animals. In certain embodiments, the compounds or compositions as described herein are highly tolerable as demonstrated by having no increase of ALT or AST over control treated animals. In certain embodiments, the compounds or compositions as described herein are highly tolerable as demonstrated by having no increase in liver, spleen, or kidney weight over control animals.
Certain embodiments provide a composition comprising or consisting essentially of the compound of any of the aforementioned embodiments or salt thereof.
Certain embodiments provide a composition comprising or consisting essentially of the compound of any of the aforementioned embodiments or salt thereof and at least one of a pharmaceutically acceptable carrier or diluent. Therefore, in one embodiment, the composition of the invention is a pharmaceutical composition.
Certain embodiments provide a medicament comprising or consisting essentially of the compound of any of the aforementioned embodiments or salt thereof. Therefore, in one embodiment, the composition of the invention is a medicament.
As used herein, the term “consists essentially of”, with reference to a composition, pharmaceutical composition or medicament, means that the compound of the invention is the only active ingredient, therapeutic agent or agent with a biologic activity within said pharmaceutical composition or medicament.
In certain embodiments, the composition has a viscosity less than about 40 centipoise (cP), less than about 30 cP, less than about 20 cP, less than about 15 cP, less than about 10 cP, less than about 5 cP, or less than about 3 cP, or less than about 1.5 cP. In certain embodiments, the composition having any of the aforementioned viscosities comprises a compound provided herein at a concentration of about 15 mg/mL, 20 mg/mL, 25 mg/mL, 50 mg/mL, 100 mg/mL, 150 mg/mL, or about 200 mg/mL. In certain embodiments, the composition having any of the aforementioned viscosities and/or compound concentrations has a temperature of room temperature or about 20° C., about 21° C., about 22° C., about 23° C., about 24° C., about 25° C., about 26° C., about 27° C., about 28° C., about 29° C., or about 30° C.
Embodiment 1. A compound comprising a modified oligonucleotide 8 to 50 linked nucleosides in length having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134.
Embodiment 2. A compound comprising a modified oligonucleotide 9 to 50 linked nucleosides in length having a nucleobase sequence comprising at least 9 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134.
Embodiment 3. A compound comprising a modified oligonucleotide 10 to 50 linked nucleosides in length having a nucleobase sequence comprising at least 10 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134.
Embodiment 4. A compound comprising a modified oligonucleotide 11 to 50 linked nucleosides in length having a nucleobase sequence comprising at least 11 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134.
Embodiment 5. A compound comprising a modified oligonucleotide 12 to 50 linked nucleosides in length having a nucleobase sequence comprising at least 12, at least 13, at least 14, or at least 15 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134.
Embodiment 6. A compound comprising a modified oligonucleotide 16 to 50 linked nucleosides in length having a nucleobase sequence comprising the nucleobase sequence of any of SEQ ID NOs: 7-3134.
Embodiment 7. A compound comprising a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 7-3134.
Embodiment 8. A compound comprising a modified oligonucleotide 8 to 50 linked nucleosides in length complementary within nucleobases 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1, wherein said modified oligonucleotide is at least 85%, 90%, 95%, or 100% complementary to SEQ ID NO: 1.
Embodiment 9. A compound comprising a modified oligonucleotide 8 to 50 linked nucleosides in length having a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1, wherein the nucleobase sequence of the modified oligonucleotide is at least 85%, 90%, 95%, or 100% complementary to SEQ ID NO: 1.
Embodiment 10. A compound comprising a modified oligonucleotide 8 to 50 linked nucleosides in length complementary within nucleobases 83,573-87,287 or 87,359-90,915 of SEQ ID NO: 1, wherein said modified oligonucleotide is at least 85%, 90%, 95%, or 100% complementary to SEQ ID NO: 1.
Embodiment 11. A compound comprising a modified oligonucleotide complementary to intron 1 of a human DNM2 pre-mRNA.
Embodiment 12. A compound comprising a modified oligonucleotide complementary to intron 11 of a human DNM2 pre-mRNA.
Embodiment 13. A compound comprising a modified oligonucleotide complementary to intron 12 of a human DNM2 pre-mRNA.
Embodiment 14. A compound comprising a modified oligonucleotide complementary to intron 13 of a human DNM2 pre-mRNA.
Embodiment 15. A compound comprising a modified oligonucleotide complementary to intron 14 of a human DNM2 pre-mRNA.
Embodiment 16. A compound comprising a modified oligonucleotide complementary to exon 10 of a human DNM2 pre-mRNA or human DNM2 mRNA.
Embodiment 17. A compound comprising a modified oligonucleotide complementary to the 3′-UTR of a human DNM2 pre-mRNA or human DNM2 mRNA.
Embodiment 18. A compound comprising a modified oligonucleotide 16 to 50 linked nucleosides in length having a nucleobase sequence comprising any of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232.
Embodiment 19. A compound comprising a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232.
Embodiment 20. A compound comprising a modified oligonucleotide having a nucleobase sequence comprising SEQ ID NO: 2879.
Embodiment 21. A compound comprising a modified oligonucleotide 16 to 50 linked nucleosides in length having a nucleobase sequence comprising any of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232, wherein the modified oligonucleotide comprises: a gap segment consisting of 8-12 linked 2′-deoxynucleosides; a 5′ wing segment consisting of 1-7 linked nucleosides; and a 3′ wing segment consisting of 1-7 linked nucleosides; wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each terminal wing nucleoside comprises a modified sugar.
Embodiment 22. A compound comprising a modified oligonucleotide 16 linked nucleosides in length having a nucleobase sequence consisting of the sequence recited in SEQ ID NO: 2879, 3056, 2123, 2189, 2453, 2160, or 2232, wherein the modified oligonucleotide comprises

- a gap segment consisting of 10 linked 2′-deoxynucleosides;
- a 5′ wing segment consisting of 3 linked nucleosides; and
- a 3′ wing segment consisting of 3 linked nucleosides;
- wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment; wherein each nucleoside of each wing segment comprises a cEt sugar moiety; wherein each internucleoside linkage is a phosphorothioate linkage; and wherein each cytosine is a 5-methylcytosine.

Embodiment 23. The compound of any one of embodiments 1-22, wherein the oligonucleotide is at least 80%, 85%, 90%, 95% or 100% complementary to any of SEQ ID NO: 1, 2, 3, or 3135.
Embodiment 24. The compound of any one of embodiments 1-23, wherein the modified oligonucleotide comprises at least one modified internucleoside linkage.
Embodiment 25. The compound of embodiments 24, wherein the at least one modified internucleoside linkage is a phosphorothioate internucleoside linkage.
Embodiment 26. The compound of any one of embodiments 1-21 or 23-25, wherein the modified oligonucleotide comprises at least one bicyclic sugar.
Embodiment 27. The compound of Embodiments 26, wherein the at least one bicyclic sugar is selected from the group consisting of LNA, ENA, and cEt.
Embodiment 28. The compound of embodiment 27, wherein the at least one bicyclic sugar moiety is a cEt sugar moiety.
Embodiment 29. The compound of any one of embodiments 1-28, wherein the modified oligonucleotide comprises at least one 5-methylcytosine.
Embodiment 30. The compound of any one of embodiments 1-29, wherein the modified oligonucleotide comprises:
a gap segment consisting of linked 2′-deoxynucleosides;
a 5′ wing segment consisting of linked nucleosides; and
a 3′ wing segment consisting of linked nucleosides;
wherein the gap segment is positioned immediately adjacent to and between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar moiety.
Embodiment 31. The compound of any one of embodiments 1-30, wherein the compound is single-stranded.
Embodiment 32. The compound of any one of embodiments 1-30, wherein the compound is double-stranded.
Embodiment 33. The compound of any one of embodiments 1-32, wherein the compound comprises at least one unmodified ribosyl sugar moiety.
Embodiment 34. The compound of any one of embodiments 1-33, wherein the compound comprises at least one unmodified 2′-deoxyribosyl sugar moiety.
Embodiment 35. The compound of any one of embodiments 1-34, wherein the modified oligonucleotide consists of 10 to 30 linked nucleosides.
Embodiment 36. The compound of any one of embodiments 1-34, wherein the modified oligonucleotide consists of 12 to 30 linked nucleosides.
Embodiment 37. The compound of any one of embodiments 1-34, wherein the modified oligonucleotide consists of 15 to 30 linked nucleosides.
Embodiment 38. The compound of any one of embodiments 1-34, wherein the modified oligonucleotide consists of 16 to 20 linked nucleosides.
Embodiment 39. A compound comprising a modified oligonucleotide according to the following formula: Gks Tks Tks Tds Ads Tds Tds Ads Tds Ads Gds Gds Gds mCks Tks Tk; wherein,

Embodiment 40. The compound of any one of embodiments 1-39 comprising a conjugate group.
Embodiment 41. The compound of embodiment 40, wherein the compound consists of the modified oligonucleotide and the conjugate group.
Embodiment 42. A compound according to the following formula:
Embodiment 43. The compound of any one of embodiments 1-39 or 42, wherein the compound
consists of the
modified oligonucleotide.
Embodiment 44. A compound consisting of a pharmaceutically acceptable salt form of any one of the compounds of embodiments 1-43.
Embodiment 45. The compound of embodiment 44, wherein the pharmaceutically acceptable salt is a sodium salt.
Embodiment 46. The compound of embodiment 44, wherein the pharmaceutically acceptable salt is a potassium salt.
Embodiment 47. A pharmaceutical composition comprising the compound of any one of embodiments 1-44 and at least one pharmaceutically acceptable carrier or diluent.
Embodiment 48. A chirally enriched population of the compounds of any one of embodiments 1-46, wherein the population is enriched for modified oligonucleotides comprising at least one particular phosphorothioate internucleoside linkage having a particular stereochemical configuration.
Embodiment 49. The chirally enriched population of embodiment 48, wherein the population is enriched for modified oligonucleotides comprising at least one particular phosphorothioate internucleoside linkage having the (Sp) configuration.
Embodiment 50. The chirally enriched population of embodiment 48, wherein the population is enriched for modified oligonucleotides comprising at least one particular phosphorothioate internucleoside linkage having the (Rp) configuration.
Embodiment 51. The chirally enriched population of embodiment 48, wherein the population is enriched for modified oligonucleotides having a particular, independently selected stereochemical configuration at each phosphorothioate internucleoside linkage
Embodiment 52. The chirally enriched population of embodiment 51, wherein the population is enriched for modified oligonucleotides having the (Sp) configuration at each phosphorothioate internucleoside linkage.
Embodiment 53. The chirally enriched population of embodiment 51, wherein the population is enriched for modified oligonucleotides having the (Rp) configuration at each phosphorothioate internucleoside linkage.
Embodiment 54. The chirally enriched population of embodiment 51, wherein the population is enriched for modified oligonucleotides having the (Rp) configuration at one particular phosphorothioate internucleoside linkage and the (Sp) configuration at each of the remaining phosphorothioate internucleoside linkages.
Embodiment 55. The chirally enriched population of embodiment 48 or embodiment 51, wherein the population is enriched for modified oligonucleotides having at least 3 contiguous phosphorothioate internucleoside linkages in the Sp, Sp, and Rp configurations, in the 5′ to 3′ direction.
Embodiment 56. A chirally enriched population of the compounds of any one of embodiments 1-46, wherein all of the phosphorothioate internucleoside linkages of the modified oligonucleotide are stereorandom.
Embodiment 57. A pharmaceutical composition comprising the population of compounds of any one of embodiments 48-56 and at least one pharmaceutically acceptable diluent or carrier.
Embodiment 58. The compound of any one of embodiments 1-46, a pharmaceutical composition comprising the compound of any one of embodiments 1-46 and at least one pharmaceutically acceptable carrier or diluent, or a pharmaceutical composition comprising the population of compounds of any one of embodiments 48-56 and at least one pharmaceutically acceptable carrier or diluent, for use in therapy.
Embodiment 59. The compound or composition of embodiment 58, for use in treating, preventing, or ameliorating centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease.
Embodiment 60. A method of treating, preventing, or ameliorating a disease associated with DNM2 in an individual comprising administering to the individual any of the compounds or compositions of embodiment 1-59, thereby treating, preventing, or ameliorating the disease.
Embodiment 61. A method of treating, preventing, or ameliorating a disease associated with DNM2 in an individual comprising administering to the individual a compound comprising a modified oligonucleotide 100% complementary to an exon 10, an intron, or the 3′-UTR of a DNM2 nucleic acid transcript, thereby treating, preventing, or ameliorating the disease.
Embodiment 62. The method of embodiment 60 or 61, wherein the disease is centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease.
Embodiment 63. The method of any one of embodiments 60-62, wherein the compound is single-stranded.
Embodiment 64. The method of any one of embodiments 60-63, wherein the DNM2 nucleic acid transcript is a pre-mRNA.
Embodiment 65. The method of any one of embodiments 60-64, wherein the disease is X-linked myotubular myopathy, autosomal recessive centronuclear myopathy, or autosomal dominant centronuclear myopathy.
Embodiment 66. The method of embodiment 65, wherein the individual has at least one mutation in at least one gene selected from among MTM1, BIN1, and DNM2.
Embodiment 67. The method of any one of embodiments 60-66, wherein the administering increases body weight or muscle strength.
Embodiment 68. A method of inhibiting expression of DNM2 in a cell comprising contacting the cell with a single-stranded compound comprising a modified oligonucleotide 100% complementary to exon 10, an intron, or the 3′-UTR of a DNM2 nucleic acid transcript, thereby inhibiting expression of DNM2 in the cell.
Embodiment 69. The method of embodiment 68, wherein the cell is in the muscle of an individual.
Embodiment 70. The method of embodiment 69, wherein the individual has, or is at risk of having, a centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease.
Embodiment 71. A method of increasing body weight or muscle strength in an individual having, or at risk of having, a disease associated with DNM2 comprising administering a single-stranded compound comprising a modified oligonucleotide 100% complementary to a DNM2 nucleic acid transcript to the individual, thereby increasing body weight or muscle strength in the individual.
Embodiment 72. The method of embodiment 71, wherein the individual has, or is at risk of having, centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease.
Embodiment 73. The method of any one of embodiments 60-72, wherein the compound is the compound of any one of embodiments 1-46.
Embodiment 74. The method of any one of embodiments 60-72, wherein the compound is a member of the chirally enriched population of any one of embodiments 48-56.
Embodiment 75. The method of any one of embodiments 60-72, wherein the compound is a component of the pharmaceutical composition of embodiment 57.
Embodiment 76. The method any one of embodiments 60-67 or 69-75, wherein the compound is administered to the individual via subcutaneous injection.
Embodiment 77. The method any one of embodiments 60-67 or 69-75, wherein the compound is administered to the individual via intramuscular injection.
Embodiment 78. The method of any one of embodiments 60-67 or 69-77, wherein the compound is administered to the individual via intravenous injection.
Embodiment 79. Use of a single-stranded compound comprising a modified oligonucleotide 100% complementary to exon 10, intron 1, intron 11, intron, 12, intron 13, intron 14, or the 3′-UTR of a DNM2 nucleic acid transcript for treating, preventing, or ameliorating a disease associated with DNM2.
Embodiment 80. The use of embodiment 79, wherein the disease is centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease.
Embodiment 81. Use of the compound of any one of embodiments 1-46 or the composition of embodiment 57 for treating, preventing, or ameliorating a disease associated with DNM2.
Embodiment 82. Use of the compound of any one of embodiments 1-46 or the composition of embodiment 57 in the manufacture of a medicament for treating, preventing, or ameliorating a disease associated with DNM2.
Embodiment 83. The use of embodiment 81 or 82, wherein the disease is centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease.
Embodiment 84. The use of embodiment 80 or 83, wherein the disease is X-linked myotubular myopathy, autosomal recessive centronuclear myopathy, or autosomal dominant centronuclear myopathy.
Embodiment 85. The use of embodiment 84, wherein the disease is associated with a mutation in at least one gene selected from among MTM1, BIN1, and DNM2.
Embodiment 86. Use of the compound of any one of embodiments 1-46 or the composition of embodiment 57 in the preparation of a medicament for treating, preventing, or ameliorating a disease associated with DNM2.
Embodiment 87. The use of embodiment 86, wherein the disease is centronuclear myopathy, Duchenne Muscular Dystrophy, or Charcot-Marie Tooth disease.
Embodiment 88. The use of embodiment 87, wherein the disease is X-linked myotubular myopathy, autosomal recessive centronuclear myopathy, or autosomal dominant centronuclear myopathy.
Embodiment 89. The use of embodiment 88, wherein the disease is associated with a mutation in at least one gene selected from among MTM1, BIN1, and DNM2.

Certain Indications

Certain embodiments provided herein relate to methods of inhibiting DNM2 expression, which can be useful for treating, preventing, or ameliorating a disease associated with DNM2 in an individual, by administration of a compound that targets DNM2, such as the compound described herein. In certain diseases associated with DNM2, modest or partial inhibition of DNM2 expression is sufficient to treat, prevent, or ameliorate the disease (See, e.g., Cowling et al. J Clin. Invest. 124, 1350-1363 (2014) and Tasfaout et al. Nat. Commun. June 7; 8:15661 (2017).) In certain embodiments, the compound can be an antisense compound, oligomeric compound, or oligonucleotide complementary to DNM2, such as the ones described herein.
Examples of diseases associated with DNM2 that are treatable, preventable, and/or ameliorable with the methods provided herein include centronuclear myopathy (CNM), Charcot-Marie-Tooth disease (CMT), and Duchenne's Muscular Dystrophy (DMD). Centronuclear myopathy includes X-linked CNM (XLCNM), autosomal dominant CNM (ADCNM) and autosomal recessive CNM (ARCNM). Mutations in several genes are associated with CNM, including mutations in MTM1, BIN1, and DNM2. Other genes have been linked to a CNM-like myopathy: RYR1 encoding for the ryanodine receptor, TTN encoding Titin, CCDC78 (OMIM 614807) and the phosphoinositides phosphatase MTMR14 (called hJUMPY; OMIM 160150).
In certain embodiments, a method of treating, preventing, or ameliorating a disease associated with DNM2 in an individual comprises administering to the individual a compound comprising an antisense compound targeted to DNM2, thereby treating, preventing, or ameliorating the disease. In certain embodiments, the compound comprises an oligonucleotide complementary to a DNM2 nucleic acid transcript. In certain embodiments, the compound is a compound as described herein. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, intron 14, exon 10, or the 3′-UTR of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, or intron 14 of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; or 97,378-104,979 of SEQ ID NO: 1. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 50 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 12 to 50 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 50 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is compound number 951799, 949935, 950023, 950089, 951372, 950060, or 950132. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound. In certain embodiments, the compound is administered to the individual subcutaneously, intradermally, intranodally, intramedullary, intramuscularly, intrasternally, by intravenous (i.v.) injection, by infusion techniques or intraperitoneally. In certain embodiments, the compound is administered to the individual via subcutaneous injection. In certain embodiments, administering the compound increases or preserves body weight or muscle strength.
In certain embodiments, a method of treating, preventing, or ameliorating CNM, DMD, or CMT comprises administering to the individual a compound comprising a modified oligonucleotide complementary to a DNM2 nucleic acid, thereby treating, preventing, or ameliorating CNM, DMD, or CMT. In certain embodiments, the compound is a compound as described herein. In certain embodiments, the compound is an antisense compound targeted to DNM2. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, intron 14, exon 10, or the 3′-UTR of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, or intron 14 of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; or 97,378-104,979 of SEQ ID NO: 1. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 50 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 12 to 50 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 50 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is compound number 951799, 949935, 950023, 950089, 951372, 950060, or 950132. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound. In certain embodiments, the compound is administered to the individual subcutaneously, intradermally, intratumorally, intranodally, intramedullary, intramuscularly, intrasternally, by intravenous (i.v.) injection, by infusion techniques or intraperitoneally. In certain embodiments, the compound is administered to the individual systemically, e.g., via subcutaneous or intramuscular injection. In certain embodiments, administering the compound increases or preserves body weight or muscle strength. In certain such embodiments, time taken to rise from the floor, nine-meter walking time, or time taken to climb four stairs is decreased. In certain embodiments, ability to lift weight, distance of a 6 minute walk, or leg function grade is increased. In certain embodiments, pulmonary function or cardiac function is improved. In certain embodiments, the individual is identified as having or at risk of having a disease associated with DNM2. Examples of factors of risk of having a disease associated with DNM2 include, but are not limited to, genetic predisposition, such as, for example, a mutation in at least one gene selected from among MTM1, BIN1, and DNM2.
In certain embodiments, a method of inhibiting expression of DNM2 in an individual having, or at risk of having, a disease associated with DNM2 comprises administering to the individual a compound comprising a modified oligonucleotide complementary to a DNM2 nucleic acid, thereby inhibiting expression of DNM2 in the individual. In certain embodiments, administering the compound inhibits expression of DNM2 in skeletal muscle. In certain embodiments, the individual has, or is at risk of having CNM, DMD, or CMT. In certain embodiments, the compound comprises an antisense compound targeted to DNM2. In certain embodiments, the antisense compound comprises an oligonucleotide complementary to a DNM2 nucleic acid transcript. In certain embodiments, the compound is a compound as described herein. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, intron 14, exon 10, or the 3′-UTR of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, or intron 14 of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; or 97,378-104,979 of SEQ ID NO: 1. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 50 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 12 to 50 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 50 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is compound number 951799, 949935, 950023, 950089, 951372, 950060, or 950132. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound. In certain embodiments, the compound is administered to the individual subcutaneously, intradermally, intranodally, intramedullary, intramuscularly, intrasternally, by intravenous (i.v.) injection, by infusion techniques or intraperitoneally. In certain embodiments, the compound is administered to the individual systemically, e.g., via subcutaneous or intramuscular injection. In certain embodiments, administering the compound increases or preserves body weight or muscle strength. In certain embodiments, the individual is identified as having or at risk of having a disease associated with DNM2.
In certain embodiments, a method of inhibiting expression of DNM2 in a cell comprises contacting the cell with a compound comprising an antisense compound targeted to DNM2, thereby inhibiting expression of DNM2 in the cell. In one embodiment, the expression of DNM2 is inhibited by at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%. In one embodiment, the expression of DNM2 is inhibited by at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% as compared to an untreated or control sample. In one embodiment, the method is an in vitro method. In certain embodiments, the cell is a muscle cell. In certain embodiments, the cell is a skeletal muscle cell. In certain embodiments, the muscle cell is in an individual who has, or is at risk of having CNM, DMD, or CMT. In certain embodiments, the compound comprises an oligonucleotide complementary to a DNM2 nucleic acid transcript. In certain embodiments, the compound is a compound as described herein. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, intron 14, exon 10, or the 3′-UTR of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, or intron 14 of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; or 97,378-104,979 of SEQ ID NO: 1. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 50 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 12 to 50 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 50 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is compound number 951799, 949935, 950023, 950089, 951372, 950060, or 950132. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound.
In certain embodiments, a method of increasing or preserving body weight or muscle strength of an individual having, or at risk of having, a disease associated with DNM2 comprises administering to the individual a compound comprising an antisense compound targeted to DNM2, increasing or preserving body weight or muscle strength of the individual. In certain such embodiments, time taken to rise from the floor, nine-meter walking time, or time taken to climb four stairs is decreased. In certain embodiments, ability to lift weight, distance of a 6 minute walk, or leg function grade is increased. In certain embodiments, pulmonary function or cardiac function is improved. In certain embodiments, the individual has, or is at risk of having, CNM, DMD, or CMT. In certain embodiments, the compound comprises an oligonucleotide complementary to a DNM2 nucleic acid transcript. In certain embodiments, the compound is a compound as described herein. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, intron 14, exon 10, or the 3′-UTR of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, or intron 14 of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; or 97,378-104,979 of SEQ ID NO: 1. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 50 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 12 to 50 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 50 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is compound number 951799, 949935, 950023, 950089, 951372, 950060, or 950132. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound. In certain embodiments, the compound is administered to the individual subcutaneously, intradermally, intranodally, intramedullary, intramuscularly, intrasternally, by intravenous (i.v.) injection, by infusion techniques or intraperitoneally. In certain embodiments, the compound is administered to the individual systemically, e.g., via subcutaneous or intramuscular injection. In certain embodiments, the individual is identified as having or at risk of having a disease associated with DNM2.
Certain embodiments are drawn to a compound comprising an antisense compound targeted to DNM2 for use in treating a disease associated with DNM2. In certain embodiments, the disease is CNM, DMD, or CMT. In certain embodiments, the compound comprises an oligonucleotide complementary to a DNM2 nucleic acid transcript. In certain embodiments, the compound is a compound as described herein. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, intron 14, exon 10, or the 3′-UTR of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, or intron 14 of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; or 97,378-104,979 of SEQ ID NO: 1. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 50 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 12 to 50 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 50 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is compound number 951799, 949935, 950023, 950089, 951372, 950060, or 950132. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound. In certain embodiments, the compound is administered to the individual subcutaneously, intradermally, intranodally, intramedullary, intramuscularly, intrasternally, by intravenous (i.v.) injection, by infusion techniques or intraperitoneally. In certain embodiments, the compound is to be administered to the individual systemically, e.g., via subcutaneous or intramuscular injection.
Certain embodiments are drawn to a compound comprising an antisense compound targeted to DNM2 for use in increasing or preserving body weight or muscle strength of an individual having or at risk of having CNM, DMD, or CMT. In certain embodiments, the compound comprises an oligonucleotide complementary to a DNM2 nucleic acid transcript. In certain embodiments, the compound is a compound as described herein. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, intron 14, exon 10, or the 3′-UTR of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, or intron 14 of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; or 97,378-104,979 of SEQ ID NO: 1. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 50 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 12 to 50 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 50 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is compound number 951799, 949935, 950023, 950089, 951372, 950060, or 950132. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound.
Certain embodiments are drawn to use of the compound comprising an antisense compound targeted to DNM2 for the manufacture or preparation of a medicament for treating a disease associated with DNM2. Certain embodiments are drawn to use of the compound comprising an antisense compound targeted to DNM2 for the preparation of a medicament for treating a disease associated with DNM2. In certain embodiments, the disease is CNM, DMD, or CMT. In certain embodiments, the compound comprises an oligonucleotide complementary to a DNM2 nucleic acid transcript. In certain embodiments, the compound is a compound as described herein. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, intron 14, exon 10, or the 3′-UTR of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, or intron 14 of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; or 97,378-104,979 of SEQ ID NO: 1. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 50 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 12 to 50 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 50 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is compound number 951799, 949935, 950023, 950089, 951372, 950060, or 950132. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound.
Certain embodiments are drawn to use of a compound comprising an antisense compound targeted to DNM2 for the manufacture or preparation of a medicament for increasing or preserving body weight or muscle strength of an individual having or at risk of having CNM, DMD, or CMT. In certain embodiments, the compound comprises an oligonucleotide complementary to a DNM2 nucleic acid transcript. In certain embodiments, the compound is a compound as described herein. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, intron 14, exon 10, or the 3′-UTR of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1. In certain embodiments, the compound comprises or consists of a modified oligonucleotide complementary to intron 1, intron 11, intron 12, intron 13, or intron 14 of a DNM2 nucleic acid transcript. In certain embodiments, the modified oligonucleotide is complementary to a sequence within nucleotides 3,404-44,737; 83,573-87,287; 87,359-90,915; 90,968-97,263; or 97,378-104,979 of SEQ ID NO: 1. In certain embodiments, the compound comprises a modified oligonucleotide 8 to 50 linked nucleosides in length and having a nucleobase sequence comprising at least 8 contiguous nucleobases of any of the nucleobase sequences of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 12 to 50 linked nucleosides in length and having a nucleobase sequence comprising the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide consisting of the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a modified oligonucleotide 16 to 50 linked nucleosides in length having a nucleobase sequence comprising any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In certain embodiments, the compound comprises a modified oligonucleotide having a nucleobase sequence consisting of any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In any of the foregoing embodiments, the modified oligonucleotide can be 10 to 30 linked nucleosides in length. In certain embodiments, the compound is compound number 951799, 949935, 950023, 950089, 951372, 950060, or 950132. In any of the foregoing embodiments, the compound can be single-stranded or double-stranded. In any of the foregoing embodiments, the compound can be an antisense compound or oligomeric compound.
In any of the foregoing methods or uses, the compound can be targeted to a DNM2 nucleic acid transcript. In certain embodiments, the compound comprises or consists of a modified oligonucleotide, for example a modified oligonucleotide 8 to 50 linked nucleosides in length, 10 to 30 linked nucleosides in length, 12 to 30 linked nucleosides in length, or 20 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is at least 80%, 85%, 90%, 95% or 100% complementary to any of the nucleobase sequences recited in SEQ ID NOs: 1, 2, 3, or 3135. In certain embodiments, the modified oligonucleotide comprises at least one modified internucleoside linkage, at least one modified sugar, and at least one modified nucleobase. In certain such embodiments, the at least one modified internucleoside linkage is a phosphorothioate internucleoside linkage, the at least one modified sugar is a bicyclic sugar, and the at least one modified nucleobase is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide comprises a gap segment consisting of linked 2′-deoxynucleosides; a 5′ wing segment consisting of linked nucleosides; and a 3′ wing segment consisting of linked nucleosides, wherein the gap segment is positioned immediately adjacent to and between the 5′ wing segment and the 3′ wing segment and wherein each terminal wing nucleoside comprises a modified sugar.
In any of the foregoing embodiments, the modified oligonucleotide may be 12 to 30, 15 to 30, 15 to 25, 15 to 24, 16 to 24, 17 to 24, 18 to 24, 19 to 24, 20 to 24, 19 to 22, 20 to 22, 16 to 20, or 17 or 20 linked nucleosides in length. In certain embodiments, the modified oligonucleotide is at least 80%, 85%, 90%, 95% or 100% complementary to any of the nucleobase sequences recited in SEQ ID NOs: 1, 2, 3, or 3135. In certain embodiments, the modified oligonucleotide comprises at least one modified internucleoside linkage, at least one modified sugar, and at least one modified nucleobase. In certain embodiments, the at least one modified internucleoside linkage is a phosphorothioate internucleoside linkage, the at least one modified sugar is a bicyclic sugar, and the at least one modified nucleobase is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide comprises a gap segment consisting of linked 2′-deoxynucleosides; a 5′ wing segment consisting of linked nucleosides; and a 3′ wing segment consisting of linked nucleosides, wherein the gap segment is positioned immediately adjacent to and between the 5′ wing segment and the 3′ wing segment and wherein each terminal wing nucleoside comprises a modified sugar.
In any of the foregoing methods or uses, the compound may comprise or consist of a modified oligonucleotide 16 to 30 linked nucleosides in length and having a nucleobase sequence comprising any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232, wherein the modified oligonucleotide comprises:

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.
In any of the foregoing methods or uses, the compound may comprise or consist of a modified oligonucleotide 16 to 30 linked nucleosides in length and having a nucleobase sequence comprising any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232, wherein the modified oligonucleotide comprises:

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment, wherein each terminal wing nucleoside comprises a modified sugar.
In any of the foregoing methods or uses, the compound may comprise or consist of a modified oligonucleotide 16 to 50 linked nucleobases in length having a nucleobase sequence comprising or consisting of the sequence recited in any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232, wherein the modified oligonucleotide comprises:

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment, wherein each nucleoside of each wing segment comprises a cEt sugar; wherein each internucleoside linkage is a phosphorothioate linkage and wherein each cytosine is a 5-methylcytosine. In certain embodiments, the modified oligonucleotide consists of 16-30 linked nucleosides. In certain embodiments, the modified oligonucleotide consists of 16 linked nucleosides.
In one embodiment, in any of the foregoing methods or uses, the compound has the following chemical structure:
In one embodiment, in any of the foregoing methods or uses, the compound can be administered systemically. In certain embodiments, the compound of any of the foregoing methods or uses can be administered through injection or infusion. In certain embodiments, the compound of any of the foregoing methods or uses can be administered via subcutaneous administration, intravenous administration, intramuscular administration, intraarterial administration, intraperitoneal administration, or intracranial administration, e.g. intrathecal or intracerebroventricular administration. In certain embodiments, the compound is administered to the individual intradermally, intranodally, intramedullary or intrasternally. In certain embodiments, the compound of any of the foregoing methods or uses can be administered orally.

Certain Combinations and Combination Therapies

In certain embodiments, a first agent comprising the compound described herein is co-administered with one or more secondary agents. In certain embodiments, such second agents are designed to treat the same disease, disorder, or condition as the first agent described herein. In certain embodiments, such second agents are designed to treat a different disease, disorder, or condition as the first agent described herein. In certain embodiments, a first agent is designed to treat an undesired side effect of a second agent. In certain embodiments, second agents are co-administered with the first agent to treat an undesired effect of the first agent. In certain embodiments, such second agents are designed to treat an undesired side effect of one or more pharmaceutical compositions as described herein. In certain embodiments, second agents are co-administered with the first agent to produce a combinational effect. In certain embodiments, second agents are co-administered with the first agent to produce a synergistic effect. In certain embodiments, the co-administration of the first and second agents permits use of lower dosages than would be required to achieve a therapeutic or prophylactic effect if the agents were administered as independent therapy.
In certain embodiments, one or more compounds or compositions provided herein are co-administered with one or more secondary agents. In certain embodiments, one or more compounds or compositions provided herein and one or more secondary agents, are administered at different times. In certain embodiments, one or more compounds or compositions provided herein and one or more secondary agents, are prepared together in a single formulation. In certain embodiments, one or more compounds or compositions provided herein and one or more secondary agents, are prepared separately.
Certain embodiments are directed to the use of a compound comprising a modified oligonucleotide complementary to a DNM2 nucleic acid transcript as described herein in combination with a secondary agent. In particular embodiments such use is in a method of treating a patient suffering from CNM, DMD, or CMT, or in the preparation or manufacture of a medicament for treating CNM, DMD, or CMT.
In certain embodiments the compound comprising a modified oligonucleotide complementary to a DNM2 nucleic acid transcript as described herein and the secondary agent are used in combination treatment by administering the two agents simultaneously, separately or sequentially. In certain embodiments the two agents are formulated as a fixed dose combination product. In other embodiments the two agents are provided to the patient as separate units which can then either be taken simultaneously or serially (sequentially).

Certain Compounds

In certain embodiments, compounds described herein can be antisense compounds. In certain embodiments, the antisense compound comprises or consists of an oligomeric compound. In certain embodiments, the oligomeric compound comprises or consists of a modified oligonucleotide. In certain embodiments, the modified oligonucleotide has a nucleobase sequence complementary to that of a target nucleic acid.
In certain embodiments, a compound described herein comprises or consists of a modified oligonucleotide. In certain embodiments, the modified oligonucleotide has a nucleobase sequence complementary to that of a target nucleic acid.
In certain embodiments, a compound or antisense compound is single-stranded. Such a single-stranded compound or antisense compound comprises or consists of an oligomeric compound. In certain embodiments, such an oligomeric compound comprises or consists of an oligonucleotide and optionally a conjugate group. In certain embodiments, the oligonucleotide is an antisense oligonucleotide. In certain embodiments, the oligonucleotide is modified. In certain embodiments, the oligonucleotide of a single-stranded antisense compound or oligomeric compound comprises a self-complementary nucleobase sequence.
In certain embodiments, a compound or antisense compound is double-stranded. Such double-stranded compounds comprise a first oligomeric compound comprising or consisting of a first modified oligonucleotide having a region complementary to a target nucleic acid and a second oligomeric compound comprising or consisting of a second oligonucleotide having a region complementary to the first modified oligonucleotide. In certain embodiments, the first oligonucleotide is 100% complementary to the second oligonucleotide. In certain embodiments, the first and second oligonucleotides include non-complementary, overhanging nucleosides. In certain embodiments, the first modified oligonucleotide comprises unmodified ribosyl sugar moieties as those found in RNA. In such embodiments, thymine nucleobases in the first and/or second oligonucleotide are replaced by uracil nucleobases. In certain embodiments, the first and/or second oligomeric compound comprises a conjugate group. In certain embodiments, the first modified oligonucleotide is 12-30 linked nucleosides in length and the second oligonucleotide is 12-30 linked nucleosides in length. In certain embodiments, the second oligonucleotide is modified. In certain embodiments, the first modified oligonucleotide has a nucleobase sequence comprising at least 8 contiguous nucleobases of any of SEQ ID NOs: 7-3134.
Examples of single-stranded and double-stranded compounds include but are not limited to oligonucleotides, siRNAs, microRNA targeting oligonucleotides, and single-stranded RNAi compounds, such as small hairpin RNAs (shRNAs), single-stranded siRNAs (ssRNAs), and microRNA mimics.
In certain embodiments, a compound described herein has a nucleobase sequence that, when written in the 5′ to 3′ direction, comprises the reverse complement of the target segment of a target nucleic acid to which it is targeted.
In certain embodiments, a compound described herein comprises an oligonucleotide 10 to 30 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 12 to 30 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 12 to 22 linked subunits in length. In certain embodiments, compound described herein comprises an oligonucleotide 14 to 30 linked subunits in length. In certain embodiments, compound described herein comprises an oligonucleotide 14 to 20 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 15 to 30 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 15 to 20 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 16 to 30 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 16 to 20 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 17 to 30 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 17 to 20 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 18 to 30 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 18 to 21 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 18 to 20 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 20 to 30 linked subunits in length. In other words, such oligonucleotides are 12 to 30 linked subunits, 14 to 30 linked subunits, 14 to 20 subunits, 15 to 30 subunits, 15 to 20 subunits, 16 to 30 subunits, 16 to 20 subunits, 17 to 30 subunits, 17 to 20 subunits, 18 to 30 subunits, 18 to 20 subunits, 18 to 21 subunits, 20 to 30 subunits, or 12 to 22 linked subunits in length, respectively. In certain embodiments, a compound described herein comprises an oligonucleotide 14 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 16 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 17 linked subunits in length. In certain embodiments, compound described herein comprises an oligonucleotide 18 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 19 linked subunits in length. In certain embodiments, a compound described herein comprises an oligonucleotide 20 linked subunits in length. In other embodiments, a compound described herein comprises an oligonucleotide 8 to 80, 12 to 50, 13 to 30, 13 to 50, 14 to 30, 14 to 50, 15 to 30, 15 to 50, 16 to 30, 16 to 50, 17 to 30, 17 to 50, 18 to 22, 18 to 24, 18 to 30, 18 to 50, 19 to 22, 19 to 30, 19 to 50, or 20 to 30 linked subunits. In certain such embodiments, the compound described herein comprises an oligonucleotide 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, or 50 linked subunits in length, or a range defined by any two of the above values. In some embodiments the linked subunits are nucleotides, nucleosides, or nucleobases.
In certain embodiments, the compound may further comprise additional features or elements, such as a conjugate group, that are attached to the oligonucleotide. In certain embodiments, such compounds are antisense compounds. In certain embodiments, such compounds are oligomeric compounds. In embodiments where a conjugate group comprises a nucleoside (i.e. a nucleoside that links the conjugate group to the oligonucleotide), the nucleoside of the conjugate group is not counted in the length of the oligonucleotide.
In certain embodiments, compounds may be shortened or truncated. For example, a single subunit may be deleted from the 5′ end (5′ truncation), or alternatively from the 3′ end (3′ truncation). A shortened or truncated compound targeted to a DNM2 nucleic acid may have two subunits deleted from the 5′ end, or alternatively may have two subunits deleted from the 3′ end, of the compound. Alternatively, the deleted nucleosides may be dispersed throughout the compound.
When a single additional subunit is present in a lengthened compound, the additional subunit may be located at the 5′ or 3′ end of the compound. When two or more additional subunits are present, the added subunits may be adjacent to each other, for example, in a compound having two subunits added to the 5′ end (5′ addition), or alternatively to the 3′ end (3′ addition), of the compound. Alternatively, the added subunits may be dispersed throughout the compound.
It is possible to increase or decrease the length of a compound, such as an oligonucleotide, and/or introduce mismatch bases without eliminating activity (Woolf et al. Proc. Nat. Acad. Sci. USA 1992, 89:7305-7309; Gautschi et al. J Nat. Cancer Inst. March 2001, 93:463-471; Maher and Dolnick Nuc. Acid. Res. 1998, 16:3341-3358). However, seemingly small changes in oligonucleotide sequence, chemistry and motif can make large differences in one or more of the many properties required for clinical development (Seth et al. J Med. Chem. 2009, 52, 10; Egli et al. J. Am. Chem. Soc. 2011, 133, 16642).
In certain embodiments, compounds described herein are interfering RNA compounds (RNAi), which include double-stranded RNA compounds (also referred to as short-interfering RNA or siRNA) and single-stranded RNAi compounds (or ssRNA). Such compounds work at least in part through the RISC pathway to degrade and/or sequester a target nucleic acid (thus, include microRNA/microRNA-mimic compounds). As used herein, the term siRNA is meant to be equivalent to other terms used to describe nucleic acid molecules that are capable of mediating sequence specific RNAi, for example short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), short hairpin RNA (shRNA), short interfering oligonucleotide, short interfering nucleic acid, short interfering modified oligonucleotide, chemically modified siRNA, post-transcriptional gene silencing RNA (ptgsRNA), and others. In addition, as used herein, 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, or epigenetics.
In certain embodiments, a compound described herein can comprise any of the oligonucleotide sequences targeted to a DNM2 nucleic acid transcript described herein. In certain embodiments, the compound can be double-stranded. In certain embodiments, the compound comprises a first strand comprising at least an 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobase portion of any one of SEQ ID NOs: 7-3134 and a second strand. In certain embodiments, the compound can be double-stranded. In certain embodiments, the compound comprises a first strand comprising at least an 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobase portion of any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232 and a second strand. In certain embodiments, the compound can be double-stranded. In certain embodiments, the compound comprises a first strand comprising at least an 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobase portion of SEQ ID NO: 2879 and a second strand. In certain embodiments, the compound comprises a first strand comprising the nucleobase sequence of any one of SEQ ID NOs: 7-3134 and a second strand. In certain embodiments, the compound comprises a first strand comprising the nucleobase sequence of any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232 and a second strand. In certain embodiments, the compound comprises a first strand comprising the nucleobase sequence of SEQ ID NO: 2879 and a second strand. In certain embodiments, the compound comprises ribonucleotides in which the first strand has uracil (U) in place of thymine (T) in any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises (i) a first strand comprising a nucleobase sequence complementary to the site on a DNM2 nucleic acid to which any of SEQ ID NOs: 7-3134 is complementary, and (ii) a second strand. In certain embodiments, the compound comprises one or more modified nucleotides in which the 2′ position in the sugar contains a halogen (such as fluorine group; 2′-F) or contains an alkoxy group (such as a methoxy group; 2′-OMe). In certain embodiments, the compound comprises at least one 2′-F sugar modification and at least one 2′-OMe sugar modification. In certain embodiments, the at least one 2′-F sugar modification and at least one 2′-OMe sugar modification are arranged in an alternating pattern for at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobases along a strand of the dsRNA compound. In certain embodiments, the compound comprises one or more linkages between adjacent nucleotides other than a naturally-occurring phosphodiester linkage. Examples of such linkages include phosphoramide, phosphorothioate, and phosphorodithioate linkages. The compounds may also be chemically modified nucleic acid molecules as taught in U.S. Pat. No. 6,673,661. In other embodiments, the compound contains one or two capped strands, as disclosed, for example, by WO 00/63364, filed Apr. 19, 2000.
In certain embodiments, the first strand of the compound is an siRNA guide strand and the second strand of the compound is an siRNA passenger strand. In certain embodiments, the second strand of the compound is complementary to the first strand. In certain embodiments, each strand of the compound is 16, 17, 18, 19, 20, 21, 22, or 23 linked nucleosides in length. In certain embodiments, the first or second strand of the compound can comprise a conjugate group.
In certain embodiments, a compound described herein can comprise any of the oligonucleotide sequences targeted to a DNM2 nucleic acid described herein. In certain embodiments, the compound is single-stranded. In certain embodiments, such a compound is a single-stranded RNAi (ssRNAi) compound. In certain embodiments, the compound comprises at least an 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobase portion of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises at least an 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobase portion of any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In certain embodiments, the compound comprises at least an 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobase portion of SEQ ID NO: 2879. In certain embodiments, the compound comprises the nucleobase sequence of any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises the nucleobase sequence of any one of SEQ ID NOs: 2879, 3056, 2123, 2189, 2453, 2160, or 2232. In certain embodiments, the compound comprises the nucleobase sequence of SEQ ID NO: 2879. In certain embodiments, the compound comprises ribonucleotides in which uracil (U) is in place of thymine (T) in any one of SEQ ID NOs: 7-3134. In certain embodiments, the compound comprises a nucleobase sequence complementary to the site on DNM2 to which any of SEQ ID NOs: 7-3134 is targeted. In certain embodiments, the compound comprises one or more modified nucleotides in which the 2′ position in the sugar contains a halogen (such as fluorine group; 2′-F) or contains an alkoxy group (such as a methoxy group; 2′-OMe). In certain embodiments, the compound comprises at least one 2′-F sugar modification and at least one 2′-OMe sugar modification. In certain embodiments, the at least one 2′-F sugar modification and at least one 2′-OMe sugar modification are arranged in an alternating pattern for at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 contiguous nucleobases along a strand of the compound. In certain embodiments, the compound comprises one or more linkages between adjacent nucleotides other than a naturally-occurring phosphodiester linkage. Examples of such linkages include phosphoramide, phosphorothioate, and phosphorodithioate linkages. The compounds may also be chemically modified nucleic acid molecules as taught in U.S. Pat. No. 6,673,661. In other embodiments, the compound contains a capped strand, as disclosed, for example, by WO 00/63364, filed Apr. 19, 2000. In certain embodiments, the compound consists of 16, 17, 18, 19, 20, 21, 22, or 23 linked nucleosides. In certain embodiments, the compound can comprise a conjugate group.
Certain compounds described herein (e.g., modified oligonucleotides) have one or more asymmetric center and thus give rise to enantiomers, diastereomers, and other stereoisomeric configurations that may be defined, in terms of absolute stereochemistry, as (R) or (S), as α or β, such as for sugar anomers, or as (D) or (L), such as for amino acids, etc. Compounds provided herein that are drawn or described as having certain stereoisomeric configurations include only the indicated compounds. Compounds provided herein that are drawn or described with undefined stereochemistry include all such possible isomers, including their stereorandom and optically pure forms. All tautomeric forms of the compounds provided herein are included unless otherwise indicated.
The compounds described herein include variations in which one or more atoms are replaced with a non-radioactive isotope or radioactive isotope of the indicated element. For example, compounds herein that comprise hydrogen atoms encompass all possible deuterium substitutions for each of the ¹H hydrogen atoms. Isotopic substitutions encompassed by the compounds herein include but are not limited to: ²H or ³H in place of ¹H, ¹³C or ¹⁴C in place of ¹²C, ¹⁵N in place of ¹⁴N, ¹⁷O or ¹⁸O in place of ¹⁶O, and ³³S, ³⁴S, ³⁵S, or ³⁶S in place of ³²S. In certain embodiments, non-radioactive isotopic substitutions may impart new properties on the oligomeric compound that are beneficial for use as a therapeutic or research tool. In certain embodiments, radioactive isotopic substitutions may make the compound suitable for research or diagnostic purposes such as imaging.

Certain Mechanisms

In certain embodiments, compounds described herein comprise or consist of modified oligonucleotides. In certain embodiments, compounds described herein are antisense compounds. In certain embodiments, compounds comprise oligomeric compounds. In certain embodiments, compounds described herein are capable of hybridizing to a DNM2 target nucleic acid, resulting in at least one antisense activity. In certain embodiments, compounds described herein selectively affect one or more target nucleic acid. Such compounds comprise a nucleobase sequence that hybridizes to one or more target nucleic acid, resulting in one or more desired antisense activity and does not hybridize to one or more non-target nucleic acid or does not hybridize to one or more non-target nucleic acid in such a way that results in a significant undesired antisense activity.
In certain antisense activities, hybridization of a compound described herein to a target nucleic acid results in recruitment of a protein that cleaves the target nucleic acid. For example, certain compounds described herein result in RNase H mediated cleavage of the target nucleic acid. RNase H is a cellular endonuclease that cleaves the RNA strand of an RNA:DNA duplex. The DNA in such an RNA:DNA duplex need not be unmodified DNA. In certain embodiments, compounds described herein are sufficiently “DNA-like” to elicit RNase H activity. Further, in certain embodiments, one or more non-DNA-like nucleoside in the gap of a gapmer is tolerated.
In certain antisense activities, compounds described herein or a portion of the compound is loaded into an RNA-induced silencing complex (RISC), ultimately resulting in cleavage of the target nucleic acid. For example, certain compounds described herein result in cleavage of the target nucleic acid by Argonaute. Compounds that are loaded into RISC are RNAi compounds. RNAi compounds may be double-stranded (siRNA) or single-stranded (ssRNA).
Antisense activities may be observed directly or indirectly. In certain embodiments, observation or detection of an antisense activity involves observation or detection of a change in an amount of a target nucleic acid or protein encoded by such target nucleic acid, a change in the ratio of splice variants of a nucleic acid or protein, and/or a phenotypic change in a cell or animal.

Target Nucleic Acids, Target Regions and Nucleotide Sequences

In certain embodiments, compounds described herein comprise or consist of an oligonucleotide comprising a region that is complementary to a target nucleic acid. In certain embodiments, the target nucleic acid is an endogenous RNA molecule. In certain embodiments, the target nucleic acid encodes a protein. In certain such embodiments, the target nucleic acid is selected from: an mRNA and a pre-mRNA, including intronic, exonic and untranslated regions. In certain embodiments, the target RNA is an mRNA. In certain embodiments, the target nucleic acid is a pre-mRNA. In certain embodiments, a pre-mRNA and corresponding mRNA are both target nucleic acids of a single compound. In certain such embodiments, the target region is entirely within an intron of a target pre-mRNA. In certain embodiments, the target region spans an intron/exon junction. In certain embodiments, the target region is at least 50% within an intron. Target nucleic acid sequences that encode DNM2 include, without limitation, the following: RefSeq or GenBank Accession No. NC_000019.10 truncated from nucleosides 10715001 to Ser. No. 10/835,000, NM_004945.3, NM_001005361.2, or NM_001005360.2 (SEQ ID Nos: 1, 2, 3, and 3135, respectively).

Hybridization

In some embodiments, hybridization occurs between a compound disclosed herein and a DNM2 nucleic acid. The most common mechanism of hybridization involves hydrogen bonding (e.g., Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding) between complementary nucleobases of the nucleic acid molecules.
Hybridization can occur under varying conditions. Hybridization conditions are sequence-dependent and are determined by the nature and composition of the nucleic acid molecules to be hybridized.
Methods of determining whether a sequence is specifically hybridizable to a target nucleic acid are well known in the art. In certain embodiments, the compounds provided herein are specifically hybridizable with a DNM2 nucleic acid.

Complementarity

In certain embodiments, compounds described herein comprise or consist of modified oligonucleotides. In certain embodiments, compounds described herein are antisense compounds. In certain embodiments, compounds comprise oligomeric compounds. In certain embodiments, oligonucleotides complementary to a DNM2 nucleic acid comprise nucleobase that are non-complementary with the DNM2 nucleic acid, yet may be tolerated provided that the compound remains able to specifically hybridize to a target nucleic acid. Moreover, a compound may hybridize over one or more segments of a DNM2 nucleic acid such that intervening or adjacent segments are not involved in the hybridization event (e.g., a loop structure, mismatch or hairpin structure).
In certain embodiments, the compounds provided herein, or a specified portion thereof, are, are at least, or are up to 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% complementary to a DNM2 nucleic acid, a target region, target segment, or specified portion thereof. In certain embodiments, the compounds provided herein, or a specified portion thereof, are 70% to 75%, 75% to 80%, 80% to 85%, 85% to 90%, 90% to 95%, 95% to 100%, or any number in between these ranges, complementary to a DNM2 nucleic acid, a target region, target segment, or specified portion thereof. Percent complementarity of a compound with a target nucleic acid can be determined using routine methods.
For example, a compound in which 18 of 20 nucleobases of the compound are complementary to a target region, and would therefore specifically hybridize, would represent 90 percent complementarity. In this example, the remaining non-complementary nucleobases may be clustered or interspersed with complementary nucleobases and need not be contiguous to each other or to complementary nucleobases. As such, a compound which is 18 nucleobases in length having four non-complementary nucleobases which are flanked by two regions of complete complementarity with the target nucleic acid would have 77.8% overall complementarity with the target nucleic acid. Percent complementarity of a compound with a region of a target nucleic acid can be determined routinely using BLAST programs (basic local alignment search tools) and PowerBLAST programs known in the art (Altschul et al., J Mol. Biol., 1990, 215, 403 410; Zhang and Madden, Genome Res., 1997, 7, 649 656). Percent homology, sequence identity or complementarity, can be determined by, for example, the Gap program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, Madison Wis.), using default settings, which uses the algorithm of Smith and Waterman (Adv. Appl. Math., 1981, 2, 482 489).
In certain embodiments, compounds described herein, or specified portions thereof, are fully complementary (i.e. 100% complementary) to a target nucleic acid, or specified portion thereof. For example, a compound may be 100% complementary to a DNM2 nucleic acid, or a target region, or a target segment or target sequence thereof. As used herein, “fully complementary” means each nucleobase of a compound is complementary to the corresponding nucleobase of a target nucleic acid. For example, a 20 nucleobase compound is fully complementary to a target sequence that is 400 nucleobases long, so long as there is a corresponding 20 nucleobase portion of the target nucleic acid that is fully complementary to the compound. Fully complementary can also be used in reference to a specified portion of the first and/or the second nucleic acid. For example, a 20 nucleobase portion of a 30 nucleobase compound can be “fully complementary” to a target sequence that is 400 nucleobases long. The 20 nucleobase portion of the 30 nucleobase compound is fully complementary to the target sequence if the target sequence has a corresponding 20 nucleobase portion wherein each nucleobase is complementary to the 20 nucleobase portion of the compound. At the same time, the entire 30 nucleobase compound may or may not be fully complementary to the target sequence, depending on whether the remaining 10 nucleobases of the compound are also complementary to the target sequence.
In certain embodiments, compounds described herein comprise one or more mismatched nucleobases relative to the target nucleic acid. In certain such embodiments, antisense activity against the target is reduced by such mismatch, but activity against a non-target is reduced by a greater amount. Thus, in certain such embodiments selectivity of the compound is improved. In certain embodiments, the mismatch is specifically positioned within an oligonucleotide having a gapmer motif. In certain such embodiments, the mismatch is at position 1, 2, 3, 4, 5, 6, 7, or 8 from the 5′-end of the gap segment. In certain such embodiments, the mismatch is at position 9, 8, 7, 6, 5, 4, 3, 2, 1 from the 3′-end of the gap segment. In certain such embodiments, the mismatch is at position 1, 2, 3, or 4 from the 5′-end of the wing segment. In certain such embodiments, the mismatch is at position 4, 3, 2, or 1 from the 3′-end of the wing segment. In certain embodiments, the mismatch is specifically positioned within an oligonucleotide not having a gapmer motif. In certain such embodiments, the mismatch is at position 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 from the 5′-end of the oligonucleotide. In certain such embodiments, the mismatch is at position 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 from the 3′-end of the oligonucleotide.
The location of a non-complementary nucleobase may be at the 5′ end or 3′ end of the compound. Alternatively, the non-complementary nucleobase or nucleobases may be at an internal position of the compound. When two or more non-complementary nucleobases are present, they may be contiguous (i.e., linked) or non-contiguous. In one embodiment, a non-complementary nucleobase is located in the wing segment of a gapmer oligonucleotide.
In certain embodiments, compounds described herein that are, or are up to 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleobases in length comprise no more than 4, no more than 3, no more than 2, or no more than 1 non-complementary nucleobase(s) relative to a target nucleic acid, such as a DNM2 nucleic acid, or specified portion thereof.
In certain embodiments, compounds described herein that are, or are up to 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleobases in length comprise no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 non-complementary nucleobase(s) relative to a target nucleic acid, such as a DNM2 nucleic acid, or specified portion thereof.
In certain embodiments, compounds described herein also include those which are complementary to a portion (a defined number of contiguous nucleobases within a region or segment) of a target nucleic acid. In certain embodiments, the compounds, are complementary to at least an 8 nucleobase portion of a target segment. In certain embodiments, the compounds are complementary to at least a 9 nucleobase portion of a target segment. In certain embodiments, the compounds are complementary to at least a 10 nucleobase portion of a target segment. In certain embodiments, the compounds are complementary to at least an 11 nucleobase portion of a target segment. In certain embodiments, the compounds are complementary to at least a 12 nucleobase portion of a target segment. In certain embodiments, the compounds are complementary to at least a 13 nucleobase portion of a target segment. In certain embodiments, the compounds are complementary to at least a 14 nucleobase portion of a target segment. In certain embodiments, the compounds are complementary to at least a 15 nucleobase portion of a target segment. In certain embodiments, the compounds are complementary to at least a 16 nucleobase portion of a target segment. Also contemplated are compounds that are complementary to at least a 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more nucleobase portion of a target segment, or a range defined by any two of these values.

Certain Compounds

In certain embodiments, compounds described herein comprise or consist of oligonucleotides consisting of linked nucleosides. Oligonucleotides may be unmodified oligonucleotides (RNA or DNA) or may be modified oligonucleotides. Modified oligonucleotides comprise at least one modification relative to unmodified RNA or DNA (i.e., comprise at least one modified nucleoside (comprising a modified sugar moiety and/or a modified nucleobase) and/or at least one modified internucleoside linkage).
I. Modifications
A. Modified Nucleosides
Modified nucleosides comprise a modified sugar moiety or a modified nucleobase or both a modified sugar moiety and a modified nucleobase.
1. Modified Sugar Moieties
In certain embodiments, sugar moieties are non-bicyclic modified sugar moieties. In certain embodiments, modified sugar moieties are bicyclic or tricyclic sugar moieties. In certain embodiments, modified sugar moieties are sugar surrogates. Such sugar surrogates may comprise one or more substitutions corresponding to those of other types of modified sugar moieties.
In certain embodiments, modified sugar moieties are non-bicyclic modified furanosyl sugar moieties comprising one or more acyclic substituent, including but not limited to substituents at the 2′, 4′, and/or 5′ positions. In certain embodiments, the furanosyl sugar moiety is a ribosyl sugar moiety. In certain embodiments one or more acyclic substituent of non-bicyclic modified sugar moieties is branched. Examples of 2′-substituent groups suitable for non-bicyclic modified sugar moieties include but are not limited to: 2′-F, 2′-OCH₃(“OMe” or “O-methyl”), and 2′-O(CH₂)₂OCH₃(“MOE”). In certain embodiments, 2′-substituent groups are selected from among: halo, allyl, amino, azido, SH, CN, OCN, CF₃, OCF₃, O—C₁-C₁₀alkoxy, O—C₁-C₁₀substituted alkoxy, C₁-C₁₀alkyl, O—C₁-C₁₀substituted alkyl, S-alkyl, N(R_m)-alkyl, O-alkenyl, S-alkenyl, N(R_m)-alkenyl, O-alkynyl, S-alkynyl, N(R_m)-alkynyl, O-alkylenyl-O-alkyl, alkynyl, alkaryl, aralkyl, O-alkaryl, O-aralkyl, O(CH₂)₂SCH₃, O(CH₂)₂N(R_m)(R_n) or OCH₂C(═O)—N(R_m)(R_n), where each R_mand R_nis, independently, H, an amino protecting group, or substituted or unsubstituted C₁-C₁₀alkyl, and the 2′-substituent groups described in Cook et al., U.S. Pat. No. 6,531,584; Cook et al., U.S. Pat. No. 5,859,221; and Cook et al., U.S. Pat. No. 6,005,087. Certain embodiments of these 2′-substituent groups can be further substituted with one or more substituent groups independently selected from among: hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro (NO₂), thiol, thioalkoxy, thioalkyl, halogen, alkyl, aryl, alkenyl and alkynyl. Examples of 4′-substituent groups suitable for non-bicyclic modified sugar moieties include but are not limited to alkoxy (e.g., methoxy), alkyl, and those described in Manoharan et al., WO 2015/106128. Examples of 5′-substituent groups suitable for non-bicyclic modified sugar moieties include but are not limited to: 5′-methyl (R or S), 5′-vinyl, and 5′-methoxy. In certain embodiments, non-bicyclic modified sugars comprise more than one non-bridging sugar substituent, for example, 2′-F-5′-methyl sugar moieties and the modified sugar moieties and modified nucleosides described in Migawa et al., WO 2008/101157 and Rajeev et al., US2013/0203836).
In certain embodiments, a 2′-substituted nucleoside or 2′-non-bicyclic modified nucleoside comprises a sugar moiety comprising a non-bridging 2′-substituent group selected from: F, NH₂, N₃, OCF₃, OCH₃, O(CH₂)₃NH₂, CH₂CH═CH₂, OCH₂CH═CH₂, OCH₂CH₂OCH₃, O(CH₂)₂SCH₃, O(CH₂)₂₀N(R_m)(R_n), O(CH₂)₂O(CH₂)₂N(CH₃)₂, and N-substituted acetamide (OCH₂C(═O)—N(R_m)(R_n)), where each R_mand R_nis, independently, H, an amino protecting group, or substituted or unsubstituted C₁-C₁₀alkyl.
In certain embodiments, a 2′-substituted nucleoside or 2′-non-bicyclic modified nucleoside comprises a sugar moiety comprising a non-bridging 2′-substituent group selected from: F, OCF₃, OCH₃, OCH₂CH₂OCH₃, O(CH₂)₂SCH₃, O(CH₂)₂₀N(CH₃)₂, O(CH₂)₂O(CH₂)₂N(CH₃)₂, and OCH₂C(═O)—N(H)CH₃(“NMA”).
In certain embodiments, a 2′-substituted nucleoside or 2′-non-bicyclic modified nucleoside comprises a sugar moiety comprising a non-bridging 2′-substituent group selected from: F, OCH₃, and OCH₂CH₂OCH₃.
Nucleosides comprising modified sugar moieties, such as non-bicyclic modified sugar moieties, may be referred to by the position(s) of the substitution(s) on the sugar moiety of the nucleoside. For example, nucleosides comprising 2′-substituted or 2-modified sugar moieties are referred to as 2′-substituted nucleosides or 2-modified nucleosides.
Certain modified sugar moieties comprise a bridging sugar substituent that forms a second ring resulting in a bicyclic sugar moiety. In certain such embodiments, the bicyclic sugar moiety comprises a bridge between the 4′ and the 2′ furanose ring atoms. In certain such embodiments, the furanose ring is a ribose ring. Examples of such 4′ to 2′ bridging sugar substituents include but are not limited to: 4′-CH₂-2′, 4′-(CH₂)₂-2′, 4′-(CH₂)_{3-2′, 4}′-CH₂—O-2′ (“LNA”), 4′-CH₂—S-2′, 4′-(CH₂)₂—O-2′ (“ENA”), 4′- CH(CH₃)—O-2′ (referred to as “constrained ethyl” or “cEt” when in the S configuration), 4′-CH₂—O—CH₂-2′, 4′-CH₂—N(R)-2′, 4′-C—H(CH₂OCH₃)—O-2′ (“constrained MOE” or “cMOE”) and analogs thereof (see, e.g., Seth et al., U.S. Pat. No. 7,399,845, Bhat et al., U.S. Pat. No. 7,569,686, Swayze et al., U.S. Pat. No. 7,741,457, and Swayze et al., U.S. Pat. No. 8,022,193), 4′-C(CH₃)(CH₃)—O-2′ and analogs thereof (see, e.g., Seth et al., U.S. Pat. No. 8,278,283), 4′-CH₂—N(OCH₃)-2′ and analogs thereof (see, e.g., Prakash et al., U.S. Pat. No. 8,278,425), 4′-CH₂—O—N(CH₃)-2′ (see, e.g., Allerson et al., U.S. Pat. No. 7,696,345 and Allerson et al., U.S. Pat. No. 8,124,745), 4′-CH₂—C(H)(CH₃)-2′ (see, e.g., Zhou, et al., J Org. Chem., 2009, 74, 118-134), 4′-CH₂—C(═CH₂)-2′ and analogs thereof (see e.g., Seth et al., U.S. Pat. No. 8,278,426), 4′-C(R_aR_b)—N(R)—O-2′, 4′-C(R_aR_b)—O—N(R)-2′, 4′-CH₂—N(R)-2′, and 4′-CH₂—N(R)—O-2′, wherein each R, R_a, and R_bis, independently, H, a protecting group, or C₁-C₁₂alkyl (see, e.g. Imanishi et al., U.S. Pat. No. 7,427,672).
In certain embodiments, such 4′ to 2′ bridges independently comprise from 1 to 4 linked groups independently selected from: —[C(R_a)(R_b)]_n—, —[C(R_a)(R_b)]_n—O—, —C(R_a)═C(R_b)—, —C(R_a)═N—, —C(═NR_a)—, —C(═O)—, —C(═S)—, —O—, —Si(R_a)₂—, —S(═O)_x—, and —N(R_a)—;
wherein:
x is 0, 1, or 2;
n is 1, 2, 3, or 4;
each R_aand R_bis, independently, H, a protecting group, hydroxyl, C₁-C₁₂alkyl, substituted C₁-C₁₂alkyl, C₂-C₁₂alkenyl, substituted C₂-C₁₂alkenyl, C₂-C₁₂alkynyl, substituted C₂-C₁₂alkynyl, C₅-C₂₀aryl, substituted C₅-C₂₀aryl, heterocycle radical, substituted heterocycle radical, heteroaryl, substituted heteroaryl, C₅-C₇alicyclic radical, substituted C₅-C₇alicyclic radical, halogen, OJ₁, NJ₁J₂, SJ₁, N₃, COOJ₁, acyl (C(═O)—H), substituted acyl, CN, sulfonyl (S(═O)₂-J₁), or sulfoxyl (S(═O)-J₁); and
each J₁and J₂is, independently, H, C₁-C₁₂alkyl, substituted C₁-C₁₂alkyl, C₂-C₁₂alkenyl, substituted C₂-C₁₂alkenyl, C₂-C₁₂alkynyl, substituted C₂-C₁₂alkynyl, C₅-C₂₀aryl, substituted C₅-C₂₀aryl, acyl (C(═O)—H), substituted acyl, a heterocycle radical, a substituted heterocycle radical, C₁-C₁₂aminoalkyl, substituted C₁-C₁₂aminoalkyl, or a protecting group.
Additional bicyclic sugar moieties are known in the art, see, for example: Freier et al., Nucleic Acids Research, 1997, 25(22), 4429-4443, Albaek et al., J Org. Chem., 2006, 71, 7731-7740, Singh et al., Chem. Commun., 1998, 4, 455-456; Koshkin et al., Tetrahedron, 1998, 54, 3607-3630; Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222; Singh et al., J Org. Chem., 1998, 63, 10035-10039; Srivastava et al., J Am. Chem. Soc., 20017, 129, 8362-8379; Elayadi et al., Wengel et a., U.S. Pat. No. 7,053,207; Imanishi et al., U.S. Pat. No. 6,268,490; Imanishi et al. U.S. Pat. No. 6,770,748; Imanishi et al., U.S. RE44,779; Wengel et al., U.S. Pat. No. 6,794,499; Wengel et al., U.S. Pat. No. 6,670,461; Wengel et al., U.S. Pat. No. 7,034,133; Wengel et al., U.S. Pat. No. 8,080,644; Wengel et al., U.S. Pat. No. 8,034,909; Wengel et al., U.S. Pat. No. 8,153,365; Wengel et al., U.S. Pat. No. 7,572,582; and Ramasamy et al., U.S. Pat. No. 6,525,191; Torsten et al., WO 2004/106356; Wengel et al., WO 1999/014226; Seth et al., WO 2007/134181; Seth et al., U.S. Pat. No. 7,547,684; Seth et al., U.S. Pat. No. 7,666,854; Seth et al., U.S. Pat. No. 8,088,746; Seth et al., U.S. Pat. No. 7,750,131; Seth et al., U.S. Pat. No. 8,030,467; Seth et al., U.S. Pat. No. 8,268,980; Seth et al., U.S. Pat. No. 8,546,556; Seth et al., U.S. Pat. No. 8,530,640; Migawa et al., U.S. Pat. No. 9,012,421; Seth et al., U.S. Pat. No. 8,501,805; and U.S. Patent Publication Nos. Allerson et al., US2008/0039618 and Migawa et al., US2015/0191727.
In certain embodiments, bicyclic sugar moieties and nucleosides incorporating such bicyclic sugar moieties are further defined by isomeric configuration. For example, an LNA nucleoside (described herein) may be in the α-L configuration or in the β-D configuration.
α-L-methyleneoxy (4′-CH₂—O-2′) or α-L-LNA bicyclic nucleosides have been incorporated into antisense oligonucleotides that showed antisense activity (Frieden et al., Nucleic Acids Research, 2003, 21, 6365-6372). Herein, general descriptions of bicyclic nucleosides include both isomeric configurations. When the positions of specific bicyclic nucleosides (e.g., LNA) are identified in exemplified embodiments herein, they are in the f-D configuration, unless otherwise specified.
In certain embodiments, modified sugar moieties comprise one or more non-bridging sugar substituent and one or more bridging sugar substituent (e.g., 5′-substituted and 4′-2′ bridged sugars).
In certain embodiments, modified sugar moieties are sugar surrogates. In certain such embodiments, the oxygen atom of the sugar moiety is replaced, e.g., with a sulfur, carbon or nitrogen atom. In certain such embodiments, such modified sugar moieties also comprise bridging and/or non-bridging substituents as described herein. For example, certain sugar surrogates comprise a 4′-sulfur atom and a substitution at the 2′-position (see, e.g., Bhat et al., U.S. Pat. No. 7,875,733 and Bhat et al., U.S. Pat. No. 7,939,677) and/or the 5′ position.
In certain embodiments, sugar surrogates comprise rings having other than 5 atoms. For example, in certain embodiments, a sugar surrogate comprises a six-membered tetrahydropyran (“THP”). Such tetrahydropyrans may be further modified or substituted. Nucleosides comprising such modified tetrahydropyrans include but are not limited to hexitol nucleic acid (“HNA”), anitol nucleic acid (“ANA”), mannitol nucleic acid (“MNA”) (see, e.g., Leumann, C J. Bioorg. & Med. Chem. 2002, 10, 841-854), fluoro HNA:
(“F-HNA”, see e.g. Swayze et al., U.S. Pat. No. 8,088,904; Swayze et al., U.S. Pat. No. 8,440,803; Swayze et al., U.S. Pat. No. 8,796,437; and Swayze et al., U.S. Pat. No. 9,005,906; F-HNA can also be referred to as a F-THP or 3′-fluoro tetrahydropyran), and nucleosides comprising additional modified THP compounds having the formula:
wherein, independently, for each of said modified THP nucleoside:
Bx is a nucleobase moiety;
T₃and T₄are each, independently, an internucleoside linking group linking the modified THP nucleoside to the remainder of an oligonucleotide or one of T₃and T₄is an internucleoside linking group linking the modified THP nucleoside to the remainder of an oligonucleotide and the other of T₃and T₄is H, a hydroxyl protecting group, a linked conjugate group, or a 5′ or 3-terminal group; q₁, q₂, q₃, q₄, q₅, q₆and q₇are each, independently, H, C₁-C₆alkyl, substituted C₁-C₆alkyl, C₂-C₆alkenyl, substituted C₂-C₆alkenyl, C₂-C₆alkynyl, or substituted C₂-C₆alkynyl; and
each of R₁and R₂is independently selected from among: hydrogen, halogen, substituted or unsubstituted alkoxy, NJ₁J₂, SJ₁, N₃, OC(═X)J₁, OC(═X)NJ₁J₂, NJ₃C(═X)NJ₁J₂, and CN, wherein X is O, S or NJ₁, and each J₁, J₂, and J₃is, independently, H or C₁-C₆alkyl.
In certain embodiments, modified THP nucleosides are provided wherein q₁, q₂, q₃, q₄, q₅, q₆and q₇are each H. In certain embodiments, at least one of q₁, q₂, q₃, q₄, q₅, q₆and q₇is other than H. In certain embodiments, at least one of q₁, q₂, q₃, q₄, q₅, q₆and q₇is methyl. In certain embodiments, modified THP nucleosides are provided wherein one of R₁and R₂is F. In certain embodiments, R₁is F and R₂is H, in certain embodiments, R₁is methoxy and R₂is H, and in certain embodiments, R₁is methoxyethoxy and R₂is H.
In certain embodiments, sugar surrogates comprise rings having more than 5 atoms and more than one heteroatom. For example, nucleosides comprising morpholino sugar moieties and their use in oligonucleotides have been reported (see, e.g., Braasch et al., Biochemistry, 2002, 41, 4503-4510 and Summerton et al., U.S. Pat. No. 5,698,685; Summerton et al., U.S. Pat. No. 5,166,315; Summerton et al., U.S. Pat. No. 5,185,444; and Summerton et al., U.S. Pat. No. 5,034,506). As used here, the term “morpholino” means a sugar surrogate having the following structure:
In certain embodiments, morpholinos may be modified, for example by adding or altering various substituent groups from the above morpholino structure. Such sugar surrogates are referred to herein as “modified morpholinos.”
In certain embodiments, sugar surrogates comprise acyclic moieties. Examples of nucleosides and oligonucleotides comprising such acyclic sugar surrogates include but are not limited to: peptide nucleic acid (“PNA”), acyclic butyl nucleic acid (see, e.g., Kumar et al., Org. Biomol. Chem., 2013, 11, 5853-5865), and nucleosides and oligonucleotides described in Manoharan et al., WO2011/133876.
Many other bicyclic and tricyclic sugar and sugar surrogate ring systems are known in the art that can be used in modified nucleosides).
2. Modified Nucleobases
In certain embodiments, modified nucleobases are selected from: 5-substituted pyrimidines, 6-azapyrimidines, alkyl or alkynyl substituted pyrimidines, alkyl substituted purines, and N-2, N-6 and O-6 substituted purines. In certain embodiments, modified nucleobases are selected from: 2-aminopropyladenine, 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-N-methylguanine, 6-N-methyladenine, 2-propyladenine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-propynyl (—C≡C—CH₃) uracil, 5-propynylcytosine, 6-azouracil, 6-azocytosine, 6-azothymine, 5-ribosyluracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl, 8-aza and other 8-substituted purines, 5-halo, particularly 5-bromo, 5-trifluoromethyl, 5-halouracil, and 5-halocytosine, 7-methylguanine, 7-methyladenine, 2-F-adenine, 2-aminoadenine, 7-deazaguanine, 7-deazaadenine, 3-deazaguanine, 3-deazaadenine, 6-N-benzoyladenine, 2-N-isobutyrylguanine, 4-N-benzoylcytosine, 4-N-benzoyluracil, 5-methyl 4-N-benzoylcytosine, 5-methyl 4-N-benzoyluracil, universal bases, hydrophobic bases, promiscuous bases, size-expanded bases, and fluorinated bases. Further modified nucleobases include tricyclic pyrimidines, such as 1,3-diazaphenoxazine-2-one, 1,3-diazaphenothiazine-2-one and 9-(2-aminoethoxy)-1,3-diazaphenoxazine-2-one (G-clamp). Modified nucleobases may also include those in which the purine or pyrimidine base is replaced with other heterocycles, for example 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine and 2-pyridone. Further nucleobases include those disclosed in Merigan et al., U.S. Pat. No. 3,687,808, those disclosed in The Concise Encyclopedia Of Polymer Science And Engineering, Kroschwitz, J. I., Ed., John Wiley & Sons, 1990, 858-859; Englisch et al., Angewandte Chemie, International Edition, 1991, 30, 613; Sanghvi, Y. S., Chapter 15, Antisense Research and Applications, Crooke, S. T. and Lebleu, B., Eds., CRC Press, 1993, 273-288; and those disclosed in Chapters 6 and 15, Antisense Drug Technology, Crooke S. T., Ed., CRC Press, 2008, 163-166 and 442-443.
Publications that teach the preparation of certain of the above noted modified nucleobases as well as other modified nucleobases include without limitation, Manohara et al., US2003/0158403; Manoharan et al., US2003/0175906; Dinh et al., U.S. Pat. No. 4,845,205; Spielvogel et al., U.S. Pat. No. 5,130,302; Rogers et al., U.S. Pat. No. 5,134,066; Bischofberger et al., U.S. Pat. No. 5,175,273; Urdea et al., U.S. Pat. No. 5,367,066; Benner et al., U.S. Pat. No. 5,432,272; Matteucci et al., U.S. Pat. No. 5,434,257; Gmeiner et al., U.S. Pat. No. 5,457,187; Cook et al., U.S. Pat. No. 5,459,255; Froehler et al., U.S. Pat. No. 5,484,908; Matteucci et al., U.S. Pat. No. 5,502,177; Hawkins et al., U.S. Pat. No. 5,525,711; Haralambidis et al., U.S. Pat. No. 5,552,540; Cook et al., U.S. Pat. No. 5,587,469; Froehler et al., U.S. Pat. No. 5,594,121; Switzer et al., U.S. Pat. No. 5,596,091; Cook et al., U.S. Pat. No. 5,614,617; Froehler et al., U.S. Pat. No. 5,645,985; Cook et al., U.S. Pat. No. 5,681,941; Cook et al., U.S. Pat. No. 5,811,534; Cook et al., U.S. Pat. No. 5,750,692; Cook et al., U.S. Pat. No. 5,948,903; Cook et al., U.S. Pat. No. 5,587,470; Cook et al., U.S. Pat. No. 5,457,191; Matteucci et al., U.S. Pat. No. 5,763,588; Froehler et al., U.S. Pat. No. 5,830,653; Cook et al., U.S. Pat. No. 5,808,027; Cook et al., 6,166,199; and Matteucci et al., U.S. Pat. No. 6,005,096.
In certain embodiments, compounds comprise or consist of a modified oligonucleotide complementary to an α-DNM2 nucleic acid comprising one or more modified nucleobases. In certain embodiments, the modified nucleobase is 5-methylcytosine. In certain embodiments, each cytosine is a 5-methylcytosine.
B. Modified Internucleoside Linkages
In certain embodiments, compounds described herein having one or more modified internucleoside linkages are selected over compounds having only phosphodiester internucleoside linkages because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for target nucleic acids, and increased stability in the presence of nucleases.
In certain embodiments, compounds comprise or consist of a modified oligonucleotide complementary to a DNM2 nucleic acid comprising one or more modified internucleoside linkages. In certain embodiments, the modified internucleoside linkages are phosphorothioate linkages. In certain embodiments, each internucleoside linkage of an antisense compound is a phosphorothioate internucleoside linkage.
In certain embodiments, nucleosides of modified oligonucleotides may be linked together using any internucleoside linkage. The two main classes of internucleoside linking groups are defined by the presence or absence of a phosphorus atom. Representative phosphorus-containing internucleoside linkages include but are not limited to phosphates, which contain a phosphodiester bond (“P═O”) (also referred to as unmodified or naturally occurring linkages), phosphotriesters, methylphosphonates, phosphoramidates, and phosphorothioates (“P═S”), and phosphorodithioates (“HS—P=S”). Representative non-phosphorus containing internucleoside linking groups include but are not limited to methylenemethylimino (—CH₂—N(CH₃)—O—CH₂—), thiodiester, thionocarbamate (—O—C(═O)(NH)—S—); siloxane (—O—SiH₂—O—); and N,N′-dimethylhydrazine (—CH₂—N(CH₃)—N(CH₃)—). Modified internucleoside linkages, compared to naturally occurring phosphate linkages, can be used to alter, typically increase, nuclease resistance of the oligonucleotide. Methods of preparation of phosphorous-containing and non-phosphorous-containing internucleoside linkages are well known to those skilled in the art.
Representative internucleoside linkages having a chiral center include but are not limited to alkylphosphonates and phosphorothioates. Modified oligonucleotides comprising internucleoside linkages having a chiral center can be prepared as populations of modified oligonucleotides comprising stereorandom internucleoside linkages, or as populations of modified oligonucleotides comprising phosphorothioate linkages in particular stereochemical configurations. In certain embodiments, populations of modified oligonucleotides comprise phosphorothioate internucleoside linkages wherein all of the phosphorothioate internucleoside linkages are stereorandom. Such modified oligonucleotides can be generated using synthetic methods that result in random selection of the stereochemical configuration of each phosphorothioate linkage. Nonetheless, as is well understood by those of skill in the art, each individual phosphorothioate of each individual oligonucleotide molecule has a defined stereoconfiguration. In certain embodiments, populations of modified oligonucleotides are enriched for modified oligonucleotides comprising one or more particular phosphorothioate internucleoside linkages in a particular, independently selected stereochemical configuration. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 65% of the molecules in the population. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 70% of the molecules in the population. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 80% of the molecules in the population. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 90% of the molecules in the population. In certain embodiments, the particular configuration of the particular phosphorothioate linkage is present in at least 99% of the molecules in the population. Such chirally enriched populations of modified oligonucleotides can be generated using synthetic methods known in the art, e.g., methods described in Oka et al., JACS 125, 8307 (2003), Wan et al. Nuc. Acid. Res. 42, 13456 (2014), and WO 2017/015555. In certain embodiments, a population of modified oligonucleotides is enriched for modified oligonucleotides having at least one indicated phosphorothioate in the (Sp) configuration. In certain embodiments, a population of modified oligonucleotides is enriched for modified oligonucleotides having at least one phosphorothioate in the (Rp) configuration. In certain embodiments, modified oligonucleotides comprising (Rp) and/or (Sp) phosphorothioates comprise one or more of the following formulas, respectively, wherein “B” indicates a nucleobase:
Unless otherwise indicated, chiral internucleoside linkages of modified oligonucleotides described herein can be stereorandom or in a particular stereochemical configuration.
Neutral internucleoside linkages include, without limitation, phosphotriesters, methylphosphonates, MMI (3′-CH₂—N(CH₃)—O-5′), amide-3 (3′-CH₂—C(═O)—N(H)-5′), amide-4 (3′-CH₂—N(H)—C(═O)-5′), formacetal (3′-O—CH₂—O-5′), methoxypropyl, and thioformacetal (3′-S—CH₂—O-5′). Further neutral internucleoside linkages include nonionic linkages comprising siloxane (dialkylsiloxane), carboxylate ester, carboxamide, sulfide, sulfonate ester and amides (See for example: Carbohydrate Modifications in Antisense Research; Y. S. Sanghvi and P. D. Cook, Eds., ACS Symposium Series 580; Chapters 3 and 4, 40-65). Further neutral internucleoside linkages include nonionic linkages comprising mixed N, O, S and CH2 component parts.
II. Certain Motifs
In certain embodiments, compounds described herein comprise or consist of oligonucleotides. Oligonucleotides can have a motif, e.g. a pattern of unmodified and/or modified sugar moieties, nucleobases, and/or internucleoside linkages. In certain embodiments, modified oligonucleotides comprise one or more modified nucleoside comprising a modified sugar. In certain embodiments, modified oligonucleotides comprise one or more modified nucleosides comprising a modified nucleobase. In certain embodiments, modified oligonucleotides comprise one or more modified internucleoside linkage. In such embodiments, the modified, unmodified, and differently modified sugar moieties, nucleobases, and/or internucleoside linkages of a modified oligonucleotide define a pattern or motif. In certain embodiments, the patterns or motifs of sugar moieties, nucleobases, and internucleoside linkages are each independent of one another. Thus, a modified oligonucleotide may be described by its sugar motif, nucleobase motif and/or internucleoside linkage motif (as used herein, nucleobase motif describes the modifications to the nucleobases independent of the sequence of nucleobases).
A. Certain Sugar Motifs
In certain embodiments, compounds described herein comprise or consist of oligonucleotides. In certain embodiments, oligonucleotides comprise one or more type of modified sugar and/or unmodified sugar moiety arranged along the oligonucleotide or region thereof in a defined pattern or sugar motif. In certain instances, such sugar motifs include but are not limited to any of the sugar modifications discussed herein.
In certain embodiments, modified oligonucleotides comprise or consist of a region having a gapmer motif, which comprises two external segments or “wings” and a central or internal segment or “gap.” The three segments of a gapmer motif (the 5′-wing, the gap, and the 3′-wing) form a contiguous sequence of nucleosides wherein at least some of the sugar moieties of the nucleosides of each of the wings differ from at least some of the sugar moieties of the nucleosides of the gap. Specifically, at least the sugar moieties of the nucleosides of each wing that are immediately adjacent to the gap (the 3′-terminal wing nucleoside of the 5′-wing and the 5′-terminal wing nucleoside of the 3′-wing) differ from the sugar moiety of the adjacent gap nucleosides. In certain embodiments, the sugar moieties within the gap are the same as one another. In certain embodiments, the gap includes one or more nucleoside having a sugar moiety that differs from the sugar moiety of one or more other nucleosides of the gap.
In certain embodiments, the wings of a gapmer each comprise 1-5 nucleosides. In certain embodiments, the wings of a gapmer each comprise 2-5 nucleosides. In certain embodiments, the wings of a gapmer each comprise 3-5 nucleosides. In certain embodiments, the nucleosides of the wings of a gapmer are all modified nucleosides. In certain such embodiments, the sugar moieties of the wings of a gapmer are all modified sugar moieties.
In certain embodiments, the gap of a gapmer comprises 7-12 nucleosides. In certain embodiments, the gap of a gapmer comprises 7-10 nucleosides. In certain embodiments, the gap of a gapmer comprises 8-10 nucleosides. In certain embodiments, the gap of a gapmer comprises 10 nucleosides. In certain embodiments, each nucleoside of the gap of a gapmer is a 2′-deoxynucleoside.
In certain embodiments, the gapmer is a deoxy gapmer. In such embodiments, the nucleosides on the gap side of each wing/gap junction are 2′-deoxynucleosides and the terminal wing nucleosides immediately adjacent to the gap comprise modified sugar moieties. In certain such embodiments, each nucleoside of the gap is a 2′-deoxynucleoside. In certain such embodiments, each nucleoside of each wing comprises a modified sugar moiety.
In certain embodiments, a modified oligonucleotide has a fully modified sugar motif wherein each nucleoside of the modified oligonucleotide comprises a modified sugar moiety. In certain embodiments, modified oligonucleotides comprise or consist of a region having a fully modified sugar motif wherein each nucleoside of the region comprises a modified sugar moiety. In certain embodiments, modified oligonucleotides comprise or consist of a region having a fully modified sugar motif, wherein each nucleoside within the fully modified region comprises the same modified sugar moiety, referred to herein as a uniformly modified sugar motif. In certain embodiments, a fully modified oligonucleotide is a uniformly modified oligonucleotide. In certain embodiments, each nucleoside of a uniformly modified oligonucleotide comprises the same 2′-modification.
B. Certain Nucleobase Motifs
In certain embodiments, compounds described herein comprise or consist of oligonucleotides. In certain embodiments, oligonucleotides comprise modified and/or unmodified nucleobases arranged along the oligonucleotide or region thereof in a defined pattern or motif. In certain embodiments, each nucleobase is modified. In certain embodiments, none of the nucleobases are modified. In certain embodiments, each purine or each pyrimidine is modified. In certain embodiments, each adenine is modified. In certain embodiments, each guanine is modified. In certain embodiments, each thymine is modified. In certain embodiments, each uracil is modified. In certain embodiments, each cytosine is modified. In certain embodiments, some or all of the cytosine nucleobases in a modified oligonucleotide are 5-methylcytosines.
In certain embodiments, modified oligonucleotides comprise a block of modified nucleobases. In certain such embodiments, the block is at the 3′-end of the oligonucleotide. In certain embodiments the block is within 3 nucleosides of the 3′-end of the oligonucleotide. In certain embodiments, the block is at the 5′-end of the oligonucleotide. In certain embodiments the block is within 3 nucleosides of the 5′-end of the oligonucleotide.
In certain embodiments, oligonucleotides having a gapmer motif comprise a nucleoside comprising a modified nucleobase. In certain such embodiments, one nucleoside comprising a modified nucleobase is in the gap of an oligonucleotide having a gapmer motif. In certain such embodiments, the sugar moiety of said nucleoside is a 2′-deoxyribosyl moiety. In certain embodiments, the modified nucleobase is selected from: a 2-thiopyrimidine and a 5-propynepyrimidine.
C. Certain Internucleoside Linkage Motifs
In certain embodiments, compounds described herein comprise or consist of oligonucleotides. In certain embodiments, oligonucleotides comprise modified and/or unmodified internucleoside linkages arranged along the oligonucleotide or region thereof in a defined pattern or motif. In certain embodiments, each internucleoside linking group is a phosphodiester internucleoside linkage (P═O). In certain embodiments, each internucleoside linking group of a modified oligonucleotide is a phosphorothioate internucleoside linkage (P═S). In certain embodiments, each internucleoside linkage of a modified oligonucleotide is independently selected from a phosphorothioate internucleoside linkage and phosphodiester internucleoside linkage. In certain embodiments, each phosphorothioate internucleoside linkage is independently selected from a stereorandom phosphorothioate, a (Sp) phosphorothioate, and a (Rp) phosphorothioate. In certain embodiments, the sugar motif of a modified oligonucleotide is a gapmer and the internucleoside linkages within the gap are all modified. In certain such embodiments, some or all of the internucleoside linkages in the wings are unmodified phosphate linkages. In certain embodiments, the terminal internucleoside linkages are modified. In certain embodiments, the sugar motif of a modified oligonucleotide is a gapmer, and the internucleoside linkage motif comprises at least one phosphodiester internucleoside linkage in at least one wing, wherein the at least one phosphodiester linkage is not a terminal internucleoside linkage, and the remaining internucleoside linkages are phosphorothioate internucleoside linkages. In certain such embodiments, all of the phosphorothioate linkages are stereorandom. In certain embodiments, all of the phosphorothioate linkages in the wings are (Sp) phosphorothioates, and the gap comprises at least one Sp, Sp, Rp motif. In certain embodiments, populations of modified oligonucleotides are enriched for modified oligonucleotides comprising such internucleoside linkage motifs.
In certain embodiments, oligonucleotides comprise a region having an alternating internucleoside linkage motif. In certain embodiments, oligonucleotides comprise a region of uniformly modified internucleoside linkages. In certain such embodiments, the internucleoside linkages are phosphorothioate internucleoside linkages. In certain embodiments, all of the internucleoside linkages of the oligonucleotide are phosphorothioate internucleoside linkages. In certain embodiments, each internucleoside linkage of the oligonucleotide is selected from phosphodiester or phosphate and phosphorothioate. In certain embodiments, each internucleoside linkage of the oligonucleotide is selected from phosphodiester or phosphate and phosphorothioate and at least one internucleoside linkage is phosphorothioate.
In certain embodiments, the oligonucleotide comprises at least 6 phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least 8 phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least 10 phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least one block of at least 6 consecutive phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least one block of at least 8 consecutive phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least one block of at least 10 consecutive phosphorothioate internucleoside linkages. In certain embodiments, the oligonucleotide comprises at least block of at least one 12 consecutive phosphorothioate internucleoside linkages. In certain such embodiments, at least one such block is located at the 3′ end of the oligonucleotide. In certain such embodiments, at least one such block is located within 3 nucleosides of the 3′ end of the oligonucleotide.
In certain embodiments, oligonucleotides comprise one or more methylphosphonate linkages. In certain embodiments, oligonucleotides having a gapmer nucleoside motif comprise a linkage motif comprising all phosphorothioate linkages except for one or two methylphosphonate linkages. In certain embodiments, one methylphosphonate linkage is in the gap of an oligonucleotide having a gapmer sugar motif.
In certain embodiments, it is desirable to arrange the number of phosphorothioate internucleoside linkages and phosphodiester internucleoside linkages to maintain nuclease resistance. In certain embodiments, it is desirable to arrange the number and position of phosphorothioate internucleoside linkages and the number and position of phosphodiester internucleoside linkages to maintain nuclease resistance. In certain embodiments, the number of phosphorothioate internucleoside linkages may be decreased and the number of phosphodiester internucleoside linkages may be increased. In certain embodiments, the number of phosphorothioate internucleoside linkages may be decreased and the number of phosphodiester internucleoside linkages may be increased while still maintaining nuclease resistance. In certain embodiments it is desirable to decrease the number of phosphorothioate internucleoside linkages while retaining nuclease resistance. In certain embodiments it is desirable to increase the number of phosphodiester internucleoside linkages while retaining nuclease resistance.
III. Certain Modified Oligonucleotides
In certain embodiments, compounds described herein comprise or consist of modified oligonucleotides. In certain embodiments, the above modifications (sugar, nucleobase, internucleoside linkage) are incorporated into a modified oligonucleotide. In certain embodiments, modified oligonucleotides are characterized by their modifications, motifs, and overall lengths. In certain embodiments, such parameters are each independent of one another. Thus, unless otherwise indicated, each internucleoside linkage of an oligonucleotide having a gapmer sugar motif may be modified or unmodified and may or may not follow the gapmer modification pattern of the sugar modifications. Likewise, such gapmer oligonucleotides may comprise one or more modified nucleobase independent of the gapmer pattern of the sugar modifications. Furthermore, in certain instances, an oligonucleotide is described by an overall length or range and by lengths or length ranges of two or more regions (e.g., a region of nucleosides having specified sugar modifications), in such circumstances it may be possible to select numbers for each range that result in an oligonucleotide having an overall length falling outside the specified range. In such circumstances, both elements must be satisfied. For example, in certain embodiments, a modified oligonucleotide consists of 15-20 linked nucleosides and has a sugar motif consisting of three regions or segments, A, B, and C, wherein region or segment A consists of 2-6 linked nucleosides having a specified sugar motif, region or segment B consists of 6-10 linked nucleosides having a specified sugar motif, and region or segment C consists of 2-6 linked nucleosides having a specified sugar motif. Such embodiments do not include modified oligonucleotides where A and C each consist of 6 linked nucleosides and B consists of 10 linked nucleosides (even though those numbers of nucleosides are permitted within the requirements for A, B, and C) because the overall length of such oligonucleotide is 22, which exceeds the upper limit of 20 for the overall length of the modified oligonucleotide. Unless otherwise indicated, all modifications are independent of nucleobase sequence except that the modified nucleobase 5-methylcytosine is necessarily a “C” in an oligonucleotide sequence.
In certain embodiments, oligonucleotides consist of X to Y linked nucleosides, where X represents the fewest number of nucleosides in the range and Y represents the largest number nucleosides in the range. In certain such embodiments, X and Y are each independently selected from 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, and 50; provided that X≤Y. For example, in certain embodiments, oligonucleotides consist of 12 to 13, 12 to 14, 12 to 15, 12 to 16, 12 to 17, 12 to 18, 12 to 19, 12 to 20, 12 to 21, 12 to 22, 12 to 23, 12 to 24, 12 to 25, 12 to 26, 12 to 27, 12 to 28, 12 to 29, 12 to 30, 13 to 14, 13 to 15, 13 to 16, 13 to 17, 13 to 18, 13 to 19, 13 to 20, 13 to 21, 13 to 22, 13 to 23, 13 to 24, 13 to 25, 13 to 26, 13 to 27, 13 to 28, 13 to 29, 13 to 30, 14 to 15, 14 to 16, 14 to 17, 14 to 18, 14 to 19, 14 to 20, 14 to 21, 14 to 22, 14 to 23, 14 to 24, 14 to 25, 14 to 26, 14 to 27, 14 to 28, 14 to 29, 14 to 30, 15 to 16, 15 to 17, 15 to 18, 15 to 19, 15 to 20, 15 to 21, 15 to 22, 15 to 23, 15 to 24, 15 to 25, 15 to 26, 15 to 27, 15 to 28, 15 to 29, 15 to 30, 16 to 17, 16 to 18, 16 to 19, 16 to 20, 16 to 21, 16 to 22, 16 to 23, 16 to 24, 16 to 25, 16 to 26, 16 to 27, 16 to 28, 16 to 29, 16 to 30, 17 to 18, 17 to 19, 17 to 20, 17 to 21, 17 to 22, 17 to 23, 17 to 24, 17 to 25, 17 to 26, 17 to 27, 17 to 28, 17 to 29, 17 to 30, 18 to 19, 18 to 20, 18 to 21, 18 to 22, 18 to 23, 18 to 24, 18 to 25, 18 to 26, 18 to 27, 18 to 28, 18 to 29, 18 to 30, 19 to 20, 19 to 21, 19 to 22, 19 to 23, 19 to 24, 19 to 25, 19 to 26, 19 to 29, 19 to 28, 19 to 29, 19 to 30, 20 to 21, 20 to 22, 20 to 23, 20 to 24, 20 to 25, 20 to 26, 20 to 27, 20 to 28, 20 to 29, 20 to 30, 21 to 22, 21 to 23, 21 to 24, 21 to 25, 21 to 26, 21 to 27, 21 to 28, 21 to 29, 21 to 30, 22 to 23, 22 to 24, 22 to 25, 22 to 26, 22 to 27, 22 to 28, 22 to 29, 22 to 30, 23 to 24, 23 to 25, 23 to 26, 23 to 27, 23 to 28, 23 to 29, 23 to 30, 24 to 25, 24 to 26, 24 to 27, 24 to 28, 24 to 29, 24 to 30, 25 to 26, 25 to 27, 25 to 28, 25 to 29, 25 to 30, 26 to 27, 26 to 28, 26 to 29, 26 to 30, 27 to 28, 27 to 29, 27 to 30, 28 to 29, 28 to 30, or 29 to 30 linked nucleosides.
In certain embodiments oligonucleotides have a nucleobase sequence that is complementary to a second oligonucleotide or an identified reference nucleic acid, such as a target nucleic acid. In certain embodiments, a region of an oligonucleotide has a nucleobase sequence that is complementary to a second oligonucleotide or an identified reference nucleic acid, such as a target nucleic acid. In certain embodiments, the nucleobase sequence of a region or entire length of an oligonucleotide is at least 70%, at least 80%, at least 90%, at least 95%, or 100% complementary to the second oligonucleotide or nucleic acid, such as a target nucleic acid.
IV. Certain Conjugated Compounds
In certain embodiments, the compounds described herein comprise or consist of an oligonucleotide (modified or unmodified) and optionally one or more conjugate groups and/or terminal groups. Conjugate groups consist of one or more conjugate moiety and a conjugate linker that links the conjugate moiety to the oligonucleotide. Conjugate groups may be attached to either or both ends of an oligonucleotide and/or at any internal position. In certain embodiments, conjugate groups are attached to the 2-position of a nucleoside of a modified oligonucleotide. In certain embodiments, conjugate groups that are attached to either or both ends of an oligonucleotide are terminal groups. In certain such embodiments, conjugate groups or terminal groups are attached at the 3′ and/or 5′-end of oligonucleotides. In certain such embodiments, conjugate groups (or terminal groups) are attached at the 3′-end of oligonucleotides. In certain embodiments, conjugate groups are attached near the 3′-end of oligonucleotides. In certain embodiments, conjugate groups (or terminal groups) are attached at the 5′-end of oligonucleotides. In certain embodiments, conjugate groups are attached near the 5′-end of oligonucleotides.
Examples of terminal groups include but are not limited to conjugate groups, capping groups, phosphate moieties, protecting groups, modified or unmodified nucleosides, and two or more nucleosides that are independently modified or unmodified.
A. Certain Conjugate Groups
In certain embodiments, oligonucleotides are covalently attached to one or more conjugate groups. In certain embodiments, conjugate groups modify one or more properties of the attached oligonucleotide, including but not limited to pharmacodynamics, pharmacokinetics, stability, binding, absorption, tissue distribution, cellular distribution, cellular uptake, charge and clearance. In certain embodiments, conjugate groups impart a new property on the attached oligonucleotide, e.g., fluorophores or reporter groups that enable detection of the oligonucleotide.
Certain conjugate groups and conjugate moieties have been described previously, for example: cholesterol moiety (Letsinger et al., Proc. Natl. Acad. Sci. USA, 1989, 86, 6553-6556), cholic acid (Manoharan et al., Bioorg. Med. Chem. Lett., 1994, 4, 1053-1060), a thioether, e.g., hexyl-S-tritylthiol (Manoharan et al., Ann. N.Y. Acad. Sci., 1992, 660, 306-309; Manoharan et al., Bioorg. Med. Chem. Lett., 1993, 3, 2765-2770), a thiocholesterol (Oberhauser et al., Nucl. Acids Res., 1992, 20, 533-538), an aliphatic chain, e.g., do-decan-diol or undecyl residues (Saison-Behmoaras et al., EMBO J., 1991, 10, 1111-1118; Kabanov et al., FEBS Lett., 1990, 259, 327-330; Svinarchuk et al., Biochimie, 1993, 75, 49-54), a phospholipid, e.g., di-hexadecyl-rac-glycerol or triethyl-ammonium 1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate (Manoharan et al., Tetrahedron Lett., 1995, 36, 3651-3654; Shea et al., Nucl. Acids Res., 1990, 18, 3777-3783), a polyamine or a polyethylene glycol chain (Manoharan et al., Nucleosides & Nucleotides, 1995, 14, 969-973), or adamantane acetic, a palmityl moiety (Mishra et al., Biochim. Biophys. Acta, 1995, 1264, 229-237), an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety (Crooke et al., J Pharmacol. Exp. Ther., 1996, i, 923-937), a tocopherol group (Nishina et al., Molecular Therapy Nucleic Acids, 2015, 4, e220; doi:10.1038/mtna.2014.72 and Nishina et al., Molecular Therapy, 2008, 16, 734-740), or a GalNAc cluster (e.g., WO2014/179620).
1. Conjugate Moieties
Conjugate moieties include, without limitation, intercalators, reporter molecules, polyamines, polyamides, peptides, carbohydrates (e.g., GalNAc), vitamin moieties, polyethylene glycols, thioethers, polyethers, cholesterols, thiocholesterols, cholic acid moieties, folate, lipids, phospholipids, biotin, phenazine, phenanthridine, anthraquinone, adamantane, acridine, fluoresceins, rhodamines, coumarins, fluorophores, and dyes.
In certain embodiments, a conjugate moiety comprises an active drug substance, for example, aspirin, warfarin, phenylbutazone, ibuprofen, suprofen, fen-bufen, ketoprofen, (S)-(+)-pranoprofen, carprofen, dansylsarcosine, 2,3,5-triiodobenzoic acid, fingolimod, flufenamic acid, folinic acid, a benzothiadiazide, chlorothiazide, a diazepine, indo-methicin, a barbiturate, a cephalosporin, a sulfa drug, an antidiabetic, an antibacterial or an antibiotic.
2. Conjugate Linkers
Conjugate moieties are attached to oligonucleotides through conjugate linkers. In certain compounds, a conjugate group is a single chemical bond (i.e. conjugate moiety is attached to an oligonucleotide via a conjugate linker through a single bond). In certain embodiments, the conjugate linker comprises a chain structure, such as a hydrocarbyl chain, or an oligomer of repeating units such as ethylene glycol, nucleosides, or amino acid units.
In certain embodiments, a conjugate linker comprises one or more groups selected from alkyl, amino, oxo, amide, disulfide, polyethylene glycol, ether, thioether, and hydroxylamino. In certain such embodiments, the conjugate linker comprises groups selected from alkyl, amino, oxo, amide and ether groups. In certain embodiments, the conjugate linker comprises groups selected from alkyl and amide groups. In certain embodiments, the conjugate linker comprises groups selected from alkyl and ether groups. In certain embodiments, the conjugate linker comprises at least one phosphorus moiety. In certain embodiments, the conjugate linker comprises at least one phosphate group. In certain embodiments, the conjugate linker includes at least one neutral linking group.
In certain embodiments, conjugate linkers, including the conjugate linkers described above, are bifunctional linking moieties, e.g., those known in the art to be useful for attaching conjugate groups to parent compounds, such as the oligonucleotides provided herein. In general, a bifunctional linking moiety comprises at least two functional groups. One of the functional groups is selected to bind to a particular site on a compound and the other is selected to bind to a conjugate group. Examples of functional groups used in a bifunctional linking moiety include but are not limited to electrophiles for reacting with nucleophilic groups and nucleophiles for reacting with electrophilic groups. In certain embodiments, bifunctional linking moieties comprise one or more groups selected from amino, hydroxyl, carboxylic acid, thiol, alkyl, alkenyl, and alkynyl.
Examples of conjugate linkers include but are not limited to pyrrolidine, 8-amino-3,6-dioxaoctanoic acid (ADO), succinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC) and 6-aminohexanoic acid (AHEX or AHA). Other conjugate linkers include but are not limited to substituted or unsubstituted C₁-C₁₀alkyl, substituted or unsubstituted C₂-C₁₀alkenyl or substituted or unsubstituted C₂-C₁₀alkynyl, wherein a nonlimiting list of preferred substituent groups includes hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro, thiol, thioalkoxy, halogen, alkyl, aryl, alkenyl and alkynyl.
In certain embodiments, conjugate linkers comprise 1-10 linker-nucleosides. In certain embodiments, such linker-nucleosides are modified nucleosides. In certain embodiments such linker-nucleosides comprise a modified sugar moiety. In certain embodiments, linker-nucleosides are unmodified. In certain embodiments, linker-nucleosides comprise an optionally protected heterocyclic base selected from a purine, substituted purine, pyrimidine or substituted pyrimidine. In certain embodiments, a cleavable moiety is a nucleoside selected from uracil, thymine, cytosine, 4-N-benzoylcytosine, 5-methylcytosine, 4-N-benzoyl-5-methylcytosine, adenine, 6-N-benzoyladenine, guanine and 2-N-isobutyrylguanine. It is typically desirable for linker-nucleosides to be cleaved from the compound after it reaches a target tissue. Accordingly, linker-nucleosides are typically linked to one another and to the remainder of the compound through cleavable bonds. In certain embodiments, such cleavable bonds are phosphodiester bonds.
Herein, linker-nucleosides are not considered to be part of the oligonucleotide. Accordingly, in embodiments in which a compound comprises an oligonucleotide consisting of a specified number or range of linked nucleosides and/or a specified percent complementarity to a reference nucleic acid and the compound also comprises a conjugate group comprising a conjugate linker comprising linker-nucleosides, those linker-nucleosides are not counted toward the length of the oligonucleotide and are not used in determining the percent complementarity of the oligonucleotide for the reference nucleic acid. For example, a compound may comprise (1) a modified oligonucleotide consisting of 8-30 nucleosides and (2) a conjugate group comprising 1-10 linker-nucleosides that are contiguous with the nucleosides of the modified oligonucleotide. The total number of contiguous linked nucleosides in such a compound is more than 30. Alternatively, an compound may comprise a modified oligonucleotide consisting of 8-30 nucleosides and no conjugate group. The total number of contiguous linked nucleosides in such a compound is no more than 30. Unless otherwise indicated conjugate linkers comprise no more than 10 linker-nucleosides. In certain embodiments, conjugate linkers comprise no more than 5 linker-nucleosides. In certain embodiments, conjugate linkers comprise no more than 3 linker-nucleosides. In certain embodiments, conjugate linkers comprise no more than 2 linker-nucleosides. In certain embodiments, conjugate linkers comprise no more than 1 linker-nucleoside.
In certain embodiments, it is desirable for a conjugate group to be cleaved from the oligonucleotide. For example, in certain circumstances compounds comprising a particular conjugate moiety are better taken up by a particular cell type, but once the compound has been taken up, it is desirable that the conjugate group be cleaved to release the unconjugated or parent oligonucleotide. Thus, certain conjugate may comprise one or more cleavable moieties, typically within the conjugate linker. In certain embodiments, a cleavable moiety is a cleavable bond. In certain embodiments, a cleavable moiety is a group of atoms comprising at least one cleavable bond. In certain embodiments, a cleavable moiety comprises a group of atoms having one, two, three, four, or more than four cleavable bonds. In certain embodiments, a cleavable moiety is selectively cleaved inside a cell or subcellular compartment, such as a lysosome. In certain embodiments, a cleavable moiety is selectively cleaved by endogenous enzymes, such as nucleases.
In certain embodiments, a cleavable bond is selected from among: an amide, an ester, an ether, one or both esters of a phosphodiester, a phosphate ester, a carbamate, or a disulfide. In certain embodiments, a cleavable bond is one or both of the esters of a phosphodiester. In certain embodiments, a cleavable moiety comprises a phosphate or phosphodiester. In certain embodiments, the cleavable moiety is a phosphate linkage between an oligonucleotide and a conjugate moiety or conjugate group.
In certain embodiments, a cleavable moiety comprises or consists of one or more linker-nucleosides. In certain such embodiments, one or more linker-nucleosides are linked to one another and/or to the remainder of the compound through cleavable bonds. In certain embodiments, such cleavable bonds are unmodified phosphodiester bonds. In certain embodiments, a cleavable moiety is 2′-deoxy nucleoside that is attached to either the 3′ or 5′-terminal nucleoside of an oligonucleotide by a phosphate internucleoside linkage and covalently attached to the remainder of the conjugate linker or conjugate moiety by a phosphate or phosphorothioate linkage. In certain such embodiments, the cleavable moiety is 2′-deoxyadenosine.
3. Certain Cell-Targeting Conjugate Moieties
In certain embodiments, a conjugate group comprises a cell-targeting conjugate moiety. In certain embodiments, a conjugate group has the general formula:

- wherein n is from 1 to about 3, m is 0 when n is 1, m is 1 when n is 2 or greater, j is 1 or 0, and k is 1 or 0.

In certain embodiments, n is 1, j is 1 and k is 0. In certain embodiments, n is 1, j is 0 and k is 1. In certain embodiments, n is 1, j is 1 and k is 1. In certain embodiments, n is 2, j is 1 and k is 0. In certain embodiments, n is 2, j is 0 and k is 1. In certain embodiments, n is 2, j is 1 and k is 1. In certain embodiments, n is 3, j is 1 and k is 0. In certain embodiments, n is 3, j is 0 and k is 1. In certain embodiments, n is 3, j is 1 and k is 1.
In certain embodiments, conjugate groups comprise cell-targeting moieties that have at least one tethered ligand. In certain embodiments, cell-targeting moieties comprise two tethered ligands covalently attached to a branching group. In certain embodiments, cell-targeting moieties comprise three tethered ligands covalently attached to a branching group.
In certain embodiments, the cell-targeting moiety comprises a branching group comprising one or more groups selected from alkyl, amino, oxo, amide, disulfide, polyethylene glycol, ether, thioether and hydroxylamino groups. In certain embodiments, the branching group comprises a branched aliphatic group comprising groups selected from alkyl, amino, oxo, amide, disulfide, polyethylene glycol, ether, thioether and hydroxylamino groups. In certain such embodiments, the branched aliphatic group comprises groups selected from alkyl, amino, oxo, amide and ether groups. In certain such embodiments, the branched aliphatic group comprises groups selected from alkyl, amino and ether groups. In certain such embodiments, the branched aliphatic group comprises groups selected from alkyl and ether groups. In certain embodiments, the branching group comprises a mono or polycyclic ring system.
In certain embodiments, each tether of a cell-targeting moiety comprises one or more groups selected from alkyl, substituted alkyl, ether, thioether, disulfide, amino, oxo, amide, phosphodiester, and polyethylene glycol, in any combination. In certain embodiments, each tether is a linear aliphatic group comprising one or more groups selected from alkyl, ether, thioether, disulfide, amino, oxo, amide, and polyethylene glycol, in any combination. In certain embodiments, each tether is a linear aliphatic group comprising one or more groups selected from alkyl, phosphodiester, ether, amino, oxo, and amide, in any combination. In certain embodiments, each tether is a linear aliphatic group comprising one or more groups selected from alkyl, ether, amino, oxo, and amid, in any combination. In certain embodiments, each tether is a linear aliphatic group comprising one or more groups selected from alkyl, amino, and oxo, in any combination. In certain embodiments, each tether is a linear aliphatic group comprising one or more groups selected from alkyl and oxo, in any combination. In certain embodiments, each tether is a linear aliphatic group comprising one or more groups selected from alkyl and phosphodiester, in any combination. In certain embodiments, each tether comprises at least one phosphorus linking group or neutral linking group. In certain embodiments, each tether comprises a chain from about 6 to about 20 atoms in length. In certain embodiments, each tether comprises a chain from about 10 to about 18 atoms in length. In certain embodiments, each tether comprises about 10 atoms in chain length.
In certain embodiments, each ligand of a cell-targeting moiety has an affinity for at least one type of receptor on a target cell. In certain embodiments, each ligand has an affinity for at least one type of receptor on the surface of a mammalian lung cell.
In certain embodiments, each ligand of a cell-targeting moiety is a carbohydrate, carbohydrate derivative, modified carbohydrate, polysaccharide, modified polysaccharide, or polysaccharide derivative. In certain such embodiments, the conjugate group comprises a carbohydrate cluster (see, e.g., Maier et al., “Synthesis of Antisense Oligonucleotides Conjugated to a Multivalent Carbohydrate Cluster for Cellular Targeting,” Bioconjugate Chemistry, 2003, 14, 18-29, or Rensen et al., “Design and Synthesis of Novel N-Acetylgalactosamine-Terminated Glycolipids for Targeting of Lipoproteins to the Hepatic Asiaglycoprotein Receptor,” J. Med. Chem. 2004, 47, 5798-5808, which are incorporated herein by reference in their entirety).
In certain such embodiments, each ligand is an amino sugar or a thio sugar. For example, amino sugars may be selected from any number of compounds known in the art, such as sialic acid, α-D-galactosamine, β-muramic acid, 2-deoxy-2-methylamino-L-glucopyranose, 4,6-dideoxy-4-formamido-2,3-di-O-methyl-D-mannopyranose, 2-deoxy-2-sulfoamino-D-glucopyranose and N-sulfo-D-glucosamine, and N-glycoloyl-α-neuraminic acid. For example, thio sugars may be selected from 5-Thio-β-D-glucopyranose, methyl 2,3,4-tri-O-acetyl-1-thio-6-O-trityl-α-D-glucopyranoside, 4-thio-β-D-galactopyranose, and ethyl 3,4,6,7-tetra-O-acetyl-2-deoxy-1,5-dithio-α-D-gluco-heptopyranoside.
In certain embodiments compounds described herein comprise a conjugate group found in any of the following references: Lee, Carbohydr Res, 1978, 67, 509-514; Connolly et al., J Biol Chem, 1982, 257, 939-945; Pavia et al., Int J Pep Protein Res, 1983, 22, 539-548; Lee et al., Biochem, 1984, 23, 4255-4261; Lee et al., Glycoconjugate J, 1987, 4, 317-328; Toyokuni et al., Tetrahedron Lett, 1990, 31, 2673-2676; Biessen et al., J Med Chem, 1995, 38, 1538-1546; Valentijn et al., Tetrahedron, 1997, 53, 759-770; Kim et al., Tetrahedron Lett, 1997, 38, 3487-3490; Lee et al., Bioconjug Chem, 1997, 8, 762-765; Kato et al., Glycobiol, 2001, 11, 821-829; Rensen et al., J Biol Chem, 2001, 276, 37577-37584; Lee et al., Methods Enzymol, 2003, 362, 38-43; Westerlind et al., Glycoconj J, 2004, 21, 227-241; Lee et al., Bioorg Med Chem Lett, 2006, 16(19), 5132-5135; Maierhofer et al., Bioorg Med Chem, 2007, 15, 7661-7676; Khorev et al., Bioorg Med Chem, 2008, 16, 5216-5231; Lee et al., Bioorg Med Chem, 2011, 19, 2494-2500; Korilova et al., Analyt Biochem, 2012, 425, 43-46; Pujol et al., Angew Chemie Int Ed Engl, 2012, 51, 7445-7448; Biessen et al., J Med Chem, 1995, 38, 1846-1852; Sliedregt et al., J Med Chem, 1999, 42, 609-618; Rensen et al., J Med Chem, 2004, 47, 5798-5808; Rensen et al., Arterioscler Thromb Vasc Biol, 2006, 26, 169-175; van Rossenberg et al., Gene Ther, 2004, 11, 457-464; Sato et al., J Am Chem Soc, 2004, 126, 14013-14022; Lee et al., J Org Chem, 2012, 77, 7564-7571; Biessen et al., FASEB J, 2000, 14, 1784-1792; Rajur et al., Bioconjug Chem, 1997, 8, 935-940; Duff et al., Methods Enzymol, 2000, 313, 297-321; Maier et al., Bioconjug Chem, 2003, 14, 18-29; Jayaprakash et al., Org Lett, 2010, 12, 5410-5413; Manoharan, Antisense Nucleic Acid Drug Dev, 2002, 12, 103-128; Merwin et al., Bioconjug Chem, 1994, 5, 612-620; Tomiya et al., Bioorg Med Chem, 2013, 21, 5275-5281; International applications WO1998/013381; WO2011/038356; WO1997/046098; WO2008/098788; WO2004/101619; WO2012/037254; WO2011/120053; WO2011/100131; WO2011/163121; WO2012/177947; WO2013/033230; WO2013/075035; WO2012/083185; WO2012/083046; WO2009/082607; WO2009/134487; WO2010/144740; WO2010/148013; WO1997/020563; WO2010/088537; WO2002/043771; WO2010/129709; WO2012/068187; WO2009/126933; WO2004/024757; WO2010/054406; WO2012/089352; WO2012/089602; WO2013/166121; WO2013/165816; U.S. Pat. Nos. 4,751,219; 8,552,163; 6,908,903; 7,262,177; 5,994,517; 6,300,319; 8,106,022; 7,491,805; 7,491,805; 7,582,744; 8,137,695; 6,383,812; 6,525,031; 6,660,720; 7,723,509; 8,541,548; 8,344,125; 8,313,772; 8,349,308; 8,450,467; 8,501,930; 8,158,601; 7,262,177; 6,906,182; 6,620,916; 8,435,491; 8,404,862; 7,851,615; Published U.S. Patent Application Publications US2011/0097264; US2011/0097265; US2013/0004427; US2005/0164235; US2006/0148740; US2008/0281044; US2010/0240730; US2003/0119724; US2006/0183886; US2008/0206869; US2011/0269814; US2009/0286973; US2011/0207799; US2012/0136042; US2012/0165393; US2008/0281041; US2009/0203135; US2012/0035115; US2012/0095075; US2012/0101148; US2012/0128760; US2012/0157509; US2012/0230938; US2013/0109817; US2013/0121954; US2013/0178512; US2013/0236968; US2011/0123520; US2003/0077829; US2008/0108801; and US2009/0203132.

Compositions and Methods for Formulating Pharmaceutical Compositions

Compounds described herein may be admixed with pharmaceutically acceptable active or inert substances for the preparation of pharmaceutical compositions. Compositions and methods for the formulation of pharmaceutical compositions are dependent upon a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.
Certain embodiments provide pharmaceutical compositions comprising one or more compounds or a salt thereof. Certain embodiments provide pharmaceutical compositions comprising one or more compounds described herein or a salt thereof. In certain embodiments, the compounds are antisense compounds or oligomeric compounds. In certain embodiments, the compounds comprise or consist of a modified oligonucleotide. In certain such embodiments, the pharmaceutical composition comprises a suitable pharmaceutically acceptable diluent or carrier. In certain embodiments, a pharmaceutical composition comprises a sterile saline solution and one or more compound. In certain embodiments, such pharmaceutical composition consists of a sterile saline solution and one or more compound. In certain embodiments, the sterile saline is pharmaceutical grade saline. In certain embodiments, a pharmaceutical composition comprises one or more compound and sterile water. In certain embodiments, a pharmaceutical composition consists of one compound and sterile water. In certain embodiments, the sterile water is pharmaceutical grade water. In certain embodiments, a pharmaceutical composition comprises or consists of one or more compound and phosphate-buffered saline (PBS). In certain embodiments, a pharmaceutical composition consists of one or more compound and sterile PBS. In certain embodiments, the sterile PBS is pharmaceutical grade PBS. Compositions and methods for the formulation of pharmaceutical compositions are dependent upon a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.
A compound described herein complementary to a DNM2 nucleic acid can be utilized in pharmaceutical compositions by combining the compound with a suitable pharmaceutically acceptable diluent or carrier and/or additional components such that the pharmaceutical composition is suitable for injection. In certain embodiments, a pharmaceutically acceptable diluent is phosphate buffered saline. Accordingly, in one embodiment, employed in the methods described herein is a pharmaceutical composition comprising a compound complementary to a DNM2 nucleic acid and a pharmaceutically acceptable diluent. In certain embodiments, the pharmaceutically acceptable diluent is phosphate buffered saline. In certain embodiments, the compound comprises or consists of a modified oligonucleotide provided herein.
Pharmaceutical compositions comprising compounds provided herein encompass any pharmaceutically acceptable salts, esters, or salts of such esters, or any other oligonucleotide which, upon administration to an animal, including a human, is capable of providing (directly or indirectly) the biologically active metabolite or residue thereof. In certain embodiments, the compounds are antisense compounds or oligomeric compounds. In certain embodiments, the compound comprises or consists of a modified oligonucleotide. Accordingly, for example, the disclosure is also drawn to pharmaceutically acceptable salts of compounds, prodrugs, pharmaceutically acceptable salts of such prodrugs, and other bioequivalents. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.
A prodrug can include the incorporation of additional nucleosides at one or both ends of a compound which are cleaved by endogenous nucleases within the body, to form the active compound.
In certain embodiments, the compounds or compositions further comprise a pharmaceutically acceptable carrier or diluent.

EXAMPLES

The Examples below describe the screening process used to identify lead compounds targeted to DNM2. Out of over 3,000 oligonucleotides that were screened, many potent and tolerable oligonucleotides were identified, and compounds 951799, 949935, 950023, 950089, 951372, 950060, and 950132 emerged as the top lead compounds. In particular, compound 951799 exhibited the best combination of properties in terms of potency and tolerability.

Non-Limiting Disclosure and Incorporation by Reference

Although the sequence listing accompanying this filing identifies each sequence as either “RNA” or “DNA” as required, in reality, those sequences may be modified with any combination of chemical modifications. One of skill in the art will readily appreciate that such designation as “RNA” or “DNA” to describe modified oligonucleotides is, in certain instances, arbitrary. For example, an oligonucleotide comprising a nucleoside comprising a 2′-OH sugar moiety and a thymine nucleobase could be described as a DNA having an RNA sugar, or as an RNA having a DNA nucleobase.
Accordingly, nucleic acid sequences provided herein, including, but not limited to those in the sequence listing, are intended to encompass nucleic acids containing any combination of unmodified or modified RNA and/or DNA, including, but not limited to such nucleic acids having modified nucleobases. By way of further example and without limitation, an oligonucleotide having the nucleobase sequence “ATCGATCG” encompasses any oligonucleotides having such nucleobase sequence, whether modified or unmodified, including, but not limited to, such compounds comprising RNA bases, such as those having sequence “AUCGAUCG” and those having some DNA bases and some RNA bases such as “AUCGATCG” and compounds having other modified nucleobases, such as “AT^mCGAUCG,” wherein ^mC indicates a cytosine base comprising a methyl group at the 5-position.
While certain compounds, compositions and methods described herein have been described with specificity in accordance with certain embodiments, the following examples serve only to illustrate the compounds described herein and are not intended to limit the same. Each of the references recited in the present application is incorporated herein by reference in its entirety.

Example 1: Effect of Modified Oligonucleotides Complementary to Dynamin 2 In Vitro

Modified oligonucleotides complementary to one or more dynamin 2 (DNM2) nucleic acids were designed and tested for their effect on dynamin 2 mRNA expression in vitro. The modified oligonucleotides were tested in a series of experiments that had similar culture conditions.
A431 cells were cultured at a density of 10,000 cells per well and treated with 4,000 nM modified oligonucleotide via free uptake or no modified oligonucleotide for untreated controls. After approximately 24 hours, RNA was isolated from the cells and DNM2 mRNA levels were measured by quantitative real-time PCR. Human primer probe set RTS36027 (forward sequence GACCTCATGCCAAAGACCAT, designated herein as SEQ ID NO: 4; reverse sequence GTCTGCCGAGGAGTATAGGTA, designated herein as SEQ ID NO: 5; probe sequence CCTTCATCCACCACGAGCTGCT, designated herein as SEQ ID: 6) was used to measure mRNA levels. DNM2 mRNA levels were normalized to total RNA content, as measured by RIBOGREEN®. Results are presented in the tables below as normalized DNM2 mRNA level, relative to untreated control cells.
The modified oligonucleotides in the tables below each have a 3-10-3 cEt gapmer motif, wherein the central gap segment contains ten 2′-deoxynucleosides and is flanked by wing segments on the 3′ and 5′ ends, each containing three cEt nucleosides. All cytosine residues throughout each modified oligonucleotide are 5-methyl cytosines. All internucleoside linkages are phosphorothioate internucleoside linkages.
Each modified oligonucleotide listed in the table below is 100% complementary to human DNM2 nucleic acid sequence GENBANK Number NC_000019.10, truncated from 10715001 to 1083500 (designated herein as SEQ ID NO: 1) and/or GENBANK Number NM_004945.3 (designated herein as SEQ ID NO: 2). “Start Site” indicates the 5′-most nucleoside of the DNM2 nucleic acid to which the oligonucleotide is complementary. “Stop Site” indicates the 3′-most nucleoside of the DNM2 nucleic acid to which the oligonucleotide is complementary. ‘N/A’ indicates that the modified oligonucleotide is not 10000 complementary to the particular nucleic acid. Several oligonucleotides match two or more sites on a nucleic acid, as shown in the table below. As shown below, modified oligonucleotides complementary to human DNM2 inhibited human DNM2 mRNA expression.

TABLE 1

DNM2 mRNA Expression

					SEQ	SEQ
	SEQ ID:	SEQ ID:			ID: 2	ID 2:	SEQ
Compound	1 Start	1 Stop		DNM2	Start	Stop	ID
Number	Site	Site	Sequence	(%Control)	Site	Site	NO

694793	3066	3081	CTCAGGCGACACCCGA	96	14	29	7

694833	60707	60722	GATGAGGGTCAAGTTC	20	580	595	8

694838	60773	60788	CTTGATCTGGTACTCG	5	646	661	9

694852	N/A	N/A	GTCCGTAGGCCTTGGG	46	774	789	10

694853	62119	62134	ACCGATGGTCCGTAGG	76	781	796	11

695014	115999	116014	AATGGTGGGCCGGCTG	99	N/A	N/A	12

695171	82451	82466	ACACACTTCAAACTCG	7	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	9	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	5	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	6	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	7	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	8	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	6	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	6	N/A	N/A	13

907925	3055	3070	CCCGACCCGAGCGACC	96	3	18	14

907926	3057	3072	CACCCGACCCGAGCGA	77	5	20	15

907927	3059	3074	GACACCCGACCCGAGC	115	7	22	16

907928	3062	3077	GGCGACACCCGACCCG	96	10	25	17

907929	3069	3084	GTTCTCAGGCGACACC	101	17	32	18

907930	3072	3087	CCGGTTCTCAGGCGAC	95	20	35	19

907931	3075	3090	CATCCGGTTCTCAGGC	58	23	38	20

907932	3078	3093	CCTCATCCGGTTCTCA	47	26	41	21

907933	3083	3098	CGCCGCCTCATCCGGT	69	31	46	22

907934	3086	3101	GGTCGCCGCCTCATCC	65	34	49	23

907935	3089	3104	CACGGTCGCCGCCTCA	29	37	52	24

907936	3091	3106	CTCACGGTCGCCGCCT	42	39	54	25

907937	3093	3108	GCCTCACGGTCGCCGC	65	41	56	26

907938	3095	3110	CGGCCTCACGGTCGCC	106	43	58	27

907939	3098	3113	GCTCGGCCTCACGGTC	44	46	61	28

907940	3101	3116	CCGGCTCGGCCTCACG	87	49	64	29

907941	3111	3126	ACGCCCGCTCCCGGCT	100	59	74	30

907942	3123	3138	GGCCTCGGCAAGACGC	96	71	86	31

907943	3126	3141	CCGGGCCTCGGCAAGA	97	74	89	32

907944	3129	3144	CGCCCGGGCCTCGGCA	84	77	92	33

907945	3144	3159	CCGTTGCTCCCCGCCC	5	92	107	34

907946	3146	3161	AGCCGTTGCTCCCCGC	12	94	109	35

907947	3149	3164	TGTAGCCGTTGCTCCC	10	97	112	36

907948	3153	3168	CGTCTGTAGCCGTTGC	8	101	116	37

907949	3171	3186	AACGACCTGGCCCCGC	12	119	134	38

907950	3177	3192	ACCCTCAACGACCTGG	41	125	140	39

907951	3180	3195	CCGACCCTCAACGACC	26	128	143	40

907952	3182	3197	CGCCGACCCTCAACGA	54	130	145	41

907953	3186	3201	CCGCCGCCGACCCTCA	35	134	149	42

907954	3190	3205	TCGCCCGCCGCCGACC	61	138	153	43

907955	3196	3211	CGCTCCTCGCCCGCCG	52	144	159	44

907956	3198	3213	TGCGCTCCTCGCCCGC	120	146	161	45

907957	3201	3216	CCCTGCGCTCCTCGCC	32	149	164	46

907958	3204	3219	GCGCCCTGCGCTCCTC	105	152	167	47

907959	3220	3235	GCGGCCCCCGGCCCGA	90	168	183	48

907960	3302	3317	CTGGCCGATGGAGCTG	129	250	265	49

907961	3304	3319	CTCTGGCCGATGGAGC	70	252	267	50

907962	3308	3323	GCAGCTCTGGCCGATG	46	256	271	51

907963	N/A	N/A	GGGAAGGAAGTCCCGG	97	349	364	52

907964	44749	44764	ATTCCTGAACCGCGGG	89	363	378	53

907965	44751	44766	CGATTCCTGAACCGCG	47	365	380	54

907966	44753	44768	GACGATTCCTGAACCG	11	367	382	55

907967	57491	57506	GACTTGCAGTGCAAAA	6	438	453	56

907968	57493	57508	TGGACTTGCAGTGCAA	15	440	455	57

907969	57495	57510	TTTGGACTTGCAGTGC	2	442	457	58

907970	57499	57514	ACTTTTTGGACTTGCA	5	446	461	59

907971	57526	57541	CCTGCCGGACTTCATC	17	473	488	60

907972	57529	57544	TCTCCTGCCGGACTTC	13	476	491	61

907973	57532	57547	CAATCTCCTGCCGGAC	20	479	494	62

907974	57534	57549	TTCAATCTCCTGCCGG	25	481	496	63

907975	60711	60726	GGTCGATGAGGGTCAA	8	584	599	64

907976	60716	60731	CGGGAGGTCGATGAGG	66	589	604	65

907977	60718	60733	CCCGGGAGGTCGATGA	117	591	606	66

907978	60722	60737	GATACCCGGGAGGTCG	55	595	610	67

907979	60724	60739	GTGATACCCGGGAGGT	11	597	612	68

907980	60728	60743	CTTGGTGATACCCGGG	57	601	616	69

907981	60734	60749	AGGCACCTTGGTGATA	40	607	622	70

907982	60776	60791	GTCCTTGATCTGGTAC	28	649	664	71

907983	60778	60793	ATGTCCTTGATCTGGT	8	651	666	72

907984	60784	60799	AGGATCATGTCCTTGA	58	657	672	73

907985	60788	60803	CTGCAGGATCATGTCC	35	661	676	74

907986	60798	60813	GGCTGATGAACTGCAG	56	671	686	75

907987	60806	60821	GCTCTCCCGGCTGATG	40	679	694	76

907988	60809	60824	GCTGCTCTCCCGGCTG	68	682	697	77

907989	60826	60841	GTGACAGCCAGAATGA	26	699	714	78

907990	60842	60857	GTCCATGTTGGCGGGC	66	715	730	79

907991	60882	60897	CGACTTCCTTGGCCAG	68	755	770	80

907992	60884	60899	ATCGACTTCCTTGGCC	99	757	772	81

907993	N/A	N/A	TGGTCCGTAGGCCTTG	72	776	791	82

907994	62116	62131	GATGGTCCGTAGGCCT	68	778	793	83

907995	62195	62210	TCAACGGGAGCAACTT	18	857	872	84

907996	N/A	N/A	GCCTCTTCTCAACGGG	105	865	880	85

907997	N/A	N/A	TAGCCTCTTCTCAACG	17	867	882	86

907998	68005	68020	GCACGGATGTCCTTCT	8	924	939	87

907999	68008	68023	GCTGCACGGATGTCCT	25	927	942	88

908000	68013	68028	CCAGTGCTGCACGGAT	17	932	947	89

908001	68057	68072	GTGCCGGTAGGCCGGG	91	976	991	90

908002	68059	68074	ATGTGCCGGTAGGCCG	83	978	993	91

908003	68062	68077	GCCATGTGCCGGTAGG	77	981	996	92

908004	68064	68079	CGGCCATGTGCCGGTA	90	983	998	93

908005	68096	68111	CGTCTTCTGCAGATGT	26	1015	1030	94

908006	71583	71598	GCCGGCAGCGACTCCC	107	1059	1074	95

908007	71586	71601	AGGGCCGGCAGCGACT	64	1062	1077	96

908008	71589	71604	CGTAGGGCCGGCAGCG	67	1065	1080	97

908009	71603	71618	TCTGTAGTTTGCTACG	34	1079	1094	98

908010	71605	71620	GCTCTGTAGTTTGCTA	42	1081	1096	99

908011	71659	71674	GTCGGGCCGAAAGTTC	15	1135	1150	100

908012	71681	71696	CTTTGGTTTTGCGGGT	36	1157	1172	101

908013	78772	78787	GAGTGTCCACCTGATC	11	1235	1250	102

908014	78786	78801	CCCGGAGAGCTCCAGA	58	1249	1264	103

908015	78789	78804	GCCCCCGGAGAGCTCC	71	1252	1267	104

908016	78791	78806	GCGCCCCCGGAGAGCT	129	1254	1269	105

908017	78804	78819	GCGATTGATTCGGGCG	50	1267	1282	106

908018	78808	78823	AGATGCGATTGATTCG	14	1271	1286	107

908019	80422	80437	GACTCCATGGATGTTC	91	1369	1384	108

908020	N/A	N/A	GCCCGGTCCTGACTCC	54	1379	1394	109

908021	N/A	N/A	AAGCCCGGTCCTGACT	81	1381	1396	110

908022	81058	81073	GAAAAGCCCGGTCCTG	35	1384	1399	111

908023	81061	81076	GGTGAAAAGCCCGGTC	58	1387	1402	112

908024	81076	81091	GAATGCCAAGTCCGGG	43	1402	1417	113

908025	81079	81094	CTCGAATGCCAAGTCC	8	1405	1420	114

908026	81082	81097	GGCCTCGAATGCCAAG	169	1408	1423	115

908027	81103	81118	GACGACCTGCTTTTTC	4	1429	1444	116

908028	81106	81121	CTTGACGACCTGCTTT	13	1432	1447	117

908029	81107	81122	GCTTGACGACCTGCTT	14	1433	1448	118

908030	81109	81124	CAGCTTGACGACCTGC	33	1435	1450	119

908031	81111	81126	TTCAGCTTGACGACCT	5	1437	1452	120

908032	81113	81128	CTTTCAGCTTGACGAC	22	1439	1454	121

908033	81115	81130	CTCTTTCAGCTTGACG	7	1441	1456	122

908034	81137	81152	GGTCGACACATTTCAG	13	1463	1478	123

908035	81139	81154	CAGGTCGACACATTTC	2	1465	1480	124

908036	81147	81162	TGGATAACCAGGTCGA	14	1473	1488	125

908037	81160	81175	ATTGATTAGCTCCTGG	2	1486	1501	126

908038	81170	81185	GCCTAACTGTATTGAT	22	1496	1511	127

908039	81172	81187	CTGCCTAACTGTATTG	28	1498	1513	128

908040	81175	81190	ACACTGCCTAACTGTA	54	1501	1516	129

908041	81182	81197	TACTGGTACACTGCCT	4	1508	1523	130

908042	N/A	N/A	CTGAGCTTACTGGTAC	19	1515	1530	131

908043	N/A	N/A	GAACTGAGCTTACTGG	10	1518	1533	132

908044	N/A	N/A	GGTAGGAACTGAGCTT	8	1523	1538	133

908045	83499	83514	TCCTCTCGCAACCGGG	46	1539	1554	134

908046	83502	83517	GTCTCCTCTCGCAACC	34	1542	1557	135

908047	83505	83520	TCTGTCTCCTCTCGCA	11	1545	1560	136

908048	83517	83532	GTGACGATTCGCTCTG	48	1557	1572	137

908049	83522	83537	AAGTGGTGACGATTCG	5	1562	1577	138

908050	83524	83539	GTAAGTGGTGACGATT	17	1564	1579	139

908051	83536	83551	CCGTTCCCGGATGTAA	86	1576	1591	140

908052	83548	83563	CGTTCTCCCCTCCCGT	9	1588	1603	141

908053	N/A	N/A	CAGAAGAATCTGGTCC	33	1606	1621	142

908054	87290	87305	GTCGATCAGCAGAAGA	33	1615	1630	143

908055	87294	87309	CAATGTCGATCAGCAG	16	1619	1634	144

908056	87296	87311	CTCAATGTCGATCAGC	10	1621	1636	145

908057	87299	87314	CTGCTCAATGTCGATC	18	1624	1639	146

908058	87302	87317	GGACTGCTCAATGTCG	58	1627	1642	147

908059	87304	87319	TAGGACTGCTCAATGT	3	1629	1644	148

908060	87308	87323	GATGTAGGACTGCTCA	19	1633	1648	149

908061	87311	87326	GTTGATGTAGGACTGC	3	1636	1651	150

908062	97288	97303	GGCTGATGTTGTTGAT	31	1760	1775	151

908063	97290	97305	CAGGCTGATGTTGTTG	36	1762	1777	152

908064	97294	97309	TCATCAGGCTGATGTT	100	1766	1781	153

908065	97299	97314	GCCTTTCATCAGGCTG	89	1771	1786	154

908066	97338	97353	TGACTCGGCAGTCAGC	86	1810	1825	155

908067	97341	97356	CAGTGACTCGGCAGTC	24	1813	1828	156

908068	97345	97360	AGGACAGTGACTCGGC	17	1817	1832	157

908069	105015	105030	ATCACGGATCTTGAGG	40	1885	1900	158

908070	105019	105034	CCACATCACGGATCTT	12	1889	1904	159

908071	105020	105035	TCCACATCACGGATCT	31	1890	1905	160

908072	105022	105037	TCTCCACATCACGGAT	72	1892	1907	161

908073	105060	105075	GAAGATGGCGAAGACG	81	1930	1945	162

908074	108791	108806	CAGGTCCTTGTAGACG	60	1963	1978	163

908075	108804	108819	GCTCGATCTGCCGCAG	55	1976	1991	164

908076	108806	108821	CAGCTCGATCTGCCGC	54	1978	1993	165

908077	108857	108872	TCGGAGGAACGAGGCC	80	2029	2044	166

908078	108859	108874	GCTCGGAGGAACGAGG	67	2031	2046	167

908079	108861	108876	CAGCTCGGAGGAACGA	59	2033	2048	168

908080	108863	108878	GCCAGCTCGGAGGAAC	93	2035	2050	169

908081	N/A	N/A	GCCTGGTCCTTCTCGG	70	2058	2073	170

908082	110075	110090	AGGTGTTCTCCTGGGC	74	2090	2105	171

908083	110077	110092	GAAGGTGTTCTCCTGG	81	2092	2107	172

908084	110080	110095	GGAGAAGGTGTTCTCC	119	2095	2110	173

908085	110160	110175	ATGGACTTGTTGATGA	11	2175	2190	174

908086*	110166	110181	TCGCGGATGGACTTGT	36	2181	2196	175

908087*	110168	110183	GGTCGCGGATGGACTT	58	2183	2198	176

908088*	110171	110186	TGAGGTCGCGGATGGA	4	2186	2201	177

908089*	114042	114057	CGTGGTGGATGAAGGC	3	2243	2258	178

908090*	114069	114084	CCGAGGAGTATAGGTA	17	2270	2285	179

908091*	114072	114087	CTGCCGAGGAGTATAG	14	2273	2288	180

908092*	114074	114089	GTCTGCCGAGGAGTAT	11	2275	2290	181

908093*	114077	114092	CTGGTCTGCCGAGGAG	9	2278	2293	182

908094*	114080	114095	GCTCTGGTCTGCCGAG	39	2281	2296	183

908095	115141	115156	CTGGACACCGGTCGGC	39	2484	2499	184

908096	115144	115159	ATGCTGGACACCGGTC	47	2487	2502	185

908097	115146	115161	GTATGCTGGACACCGG	33	2489	2504	186

908098	115150	115165	GGGTGTATGCTGGACA	11	2493	2508	187

908099	115214	115229	CACGGGAACAGGAATC	25	2557	2572	188

908100	115221	115236	CTGCCCCCACGGGAAC	80	2564	2579	189

908101	115238	115253	CGCCGAGAAGGAGGCT	97	2581	2596	190

908102	115283	115298	GTTGGCAAACACGCTC	16	2626	2641	191

908103	115285	115300	CTGTTGGCAAACACGC	30	2628	2643	192

908104	115292	115307	GAGGTCACTGTTGGCA	9	2635	2650	193

908105	115324	115339	GGCCGAGATGGGATCT	73	2667	2682	194

908106	115326	115341	CTGGCCGAGATGGGAT	45	2669	2684	195

908107	115329	115344	GAACTGGCCGAGATGG	6	2672	2687	196

908108	115332	115347	TCCGAACTGGCCGAGA	33	2675	2690	197

908109	115336	115351	GGGATCCGAACTGGCC	82	2679	2694	198

908110	116010	116025	GCTGGGCGGATAATGG	45	2754	2769	199

908111	116015	116030	GCTCGGCTGGGCGGAT	101	2759	2774	200

908112	116017	116032	TGGCTCGGCTGGGCGG	89	2761	2776	201

908113	116022	116037	AGGGATGGCTCGGCTG	75	2766	2781	202

908114	116030	116045	AGTCGAGCAGGGATGG	53	2774	2789	203

908115	116035	116050	GGCCTAGTCGAGCAGG	104	2779	2794	204

908116	116038	116053	CGAGGCCTAGTCGAGC	79	2782	2797	205

908117	116042	116057	CCCTCGAGGCCTAGTC	87	2786	2801	206

908118	116045	116060	CCCCCCTCGAGGCCTA	69	2789	2804	207

908119	116047	116062	CGCCCCCCTCGAGGCC	89	2791	2806	208

908120	116057	116072	CCCGAGAGCACGCCCC	40	2801	2816	209

908121	116060	116075	CCCCCCGAGAGCACGC	38	2804	2819	210

908122	116063	116078	GGCCCCCCCGAGAGCA	95	2807	2822	211

908123	116068	116083	CGTGAGGCCCCCCCGA	67	2812	2827	212

908124	116088	116103	AAGCTCCTGCGCCGCG	55	2832	2847	213

908125	116096	116111	GACCACTGAAGCTCCT	32	2840	2855	214

908126	116098	116113	CAGACCACTGAAGCTC	38	2842	2857	215

908127	116108	116123	CGGAGGGCCCCAGACC	100	2852	2867	216

908128	116110	116125	GGCGGAGGGCCCCAGA	91	2854	2869	217

908129	116128	116143	TGGTCCCAGCATAGGG	88	2872	2887	218

908130	116130	116145	CCTGGTCCCAGCATAG	73	2874	2889	219

908131	116138	116153	ACTGGGAGCCTGGTCC	72	2882	2897	220

908132	116164	116179	CGTTAAGGAAGAGGCC	69	2908	2923	221

908133	116209	116224	GCGGTGTCCAGCCAGG	49	2953	2968	222

908134	116211	116226	GTGCGGTGTCCAGCCA	37	2955	2970	223

908135	116215	116230	CGCAGTGCGGTGTCCA	13	2959	2974	224

908136	116218	116233	TTGCGCAGTGCGGTGT	29	2962	2977	225

908137	116220	116235	CTTTGCGCAGTGCGGT	15	2964	2979	226

908138	116223	116238	CCCCTTTGCGCAGTGC	12	2967	2982	227

908139	116226	116241	GGGCCCCTTTGCGCAG	101	2970	2985	228

908140	116228	116243	CAGGGCCCCTTTGCGC	92	2972	2987	229

908141	116260	116275	TGCAACACCCCAGCGC	78	3004	3019	230

908142	116265	116280	CAAAGTGCAACACCCC	20	3009	3024	231

908143	116268	116283	CCCCAAAGTGCAACAC	47	3012	3027	232

908144	116300	116315	TGGTCCCCCCTCTGCC	64	3044	3059	233

908145	116309	116324	CAAGGGTTCTGGTCCC	45	3053	3068	234

908146	116312	116327	TGTCAAGGGTTCTGGT	15	3056	3071	235

908147	116314	116329	GGTGTCAAGGGTTCTG	8	3058	3073	236

908148	116318	116333	GGATGGTGTCAAGGGT	7	3062	3077	237

908149	116331	116346	GACCCCTCATTCAGGA	84	3075	3090	238

908150	116333	116348	TGGACCCCTCATTCAG	64	3077	3092	239

908151	116336	116351	GGCTGGACCCCTCATT	80	3080	3095	240

908152	116348	116363	GAGTCCCCCCCAGGCT	63	3092	3107	241

908153	116352	116367	GGTAGAGTCCCCCCCA	31	3096	3111	242

908154	116356	116371	CCTTGGTAGAGTCCCC	3	3100	3115	243

908155	116359	116374	AGACCTTGGTAGAGTC	73	3103	3118	244

908156	116385	116400	CCTACATGGGCTTTCC	37	3129	3144	245

908157	116389	116404	CTGCCCTACATGGGCT	115	3133	3148	246

908158	116398	116413	ATAGAAGGCCTGCCCT	71	3142	3157	247

908159	116400	116415	TTATAGAAGGCCTGCC	41	3144	3159	248

908160	116404	116419	GCACTTATAGAAGGCC	70	3148	3163	249

908161	116408	116423	GCCCGCACTTATAGAA	45	3152	3167	250

908162	116410	116425	GTGCCCGCACTTATAG	66	3154	3169	251

908163	116418	116433	CGCCCTTGGTGCCCGC	62	3162	3177	252

908164	116455	116470	TATACCCCTGCACCCC	11	3199	3214	253

908165	116464	116479	GGAAGTTGATATACCC	29	3208	3223	254

908166	116471	116486	GCTAATGGGAAGTTGA	13	3215	3230	255

908167	116474	116489	CCTGCTAATGGGAAGT	14	3218	3233	256

908168	116477	116492	GCTCCTGCTAATGGGA	139	3221	3236	257

908169	116481	116496	GGGAGCTCCTGCTAAT	102	3225	3240	258

908170	116497	116512	GCCAGGCTTGCCGCTG	80	3241	3256	259

908171	116499	116514	GGGCCAGGCTTGCCGC	87	3243	3258	260

908172	116511	116526	ACCGAGCCCACTGGGC	72	3255	3270	261

908173	116513	116528	CTACCGAGCCCACTGG	57	3257	3272	262

908174	116518	116533	GGGCACTACCGAGCCC	87	3262	3277	263

908175	116523	116538	CAGCTGGGCACTACCG	86	3267	3282	264

908176	116541	116556	TGTACACCTCAGGCCT	83	3285	3300	265

908177	116544	116559	CTATGTACACCTCAGG	10	3288	3303	266

908178	116547	116562	GGACTATGTACACCTC	7	3291	3306	267

908179	116550	116565	GAAGGACTATGTACAC	8	3294	3309	268

908180	116556	116571	GGCCGGGAAGGACTAT	51	3300	3315	269

908181	116558	116573	ATGGCCGGGAAGGACT	78	3302	3317	270

908182	116561	116576	AATATGGCCGGGAAGG	19	3305	3320	271

908183	116565	116580	GGTTAATATGGCCGGG	13	3309	3324	272

908184	116573	116588	GGCTGTGTGGTTAATA	8	3317	3332	273

908185	116603	116618	CCTCTGGCAGCCGAGG	100	3347	3362	274

908186	116606	116621	GCACCTCTGGCAGCCG	73	3350	3365	275

908187	116608	116623	AGGCACCTCTGGCAGC	61	3352	3367	276

908188	116614	116629	TAGCAAAGGCACCTCT	41	3358	3373	277

908189	116617	116632	GCCTAGCAAAGGCACC	146	3361	3376	278

908190	116621	116636	CCGGGCCTAGCAAAGG	60	3365	3380	279

908191	116624	116639	GCTCCGGGCCTAGCAA	123	3368	3383	280

908192	116626	116641	CGGCTCCGGGCCTAGC	97	3370	3385	281

908193	116630	116645	CCAACGGCTCCGGGCC	117	3374	3389	282

908194	116638	116653	GGCCCGGGCCAACGGC	109	3382	3397	283

908195	116640	116655	CCGGCCCGGGCCAACG	104	3384	3399	284

908196	116642	116657	GGCCGGCCCGGGCCAA	99	3386	3401	285

908197	116644	116659	AAGGCCGGCCCGGGCC	86	3388	3403	286

908198	116646	116661	GCAAGGCCGGCCCGGG	88	3390	3405	287

908199	116648	116663	GGGCAAGGCCGGCCCG	125	3392	3407	288

908200	116650	116665	TAGGGCAAGGCCGGCC	99	3394	3409	289

908201	116655	116670	AGGAATAGGGCAAGGC	6	3399	3414	290

908202	116712	116727	ACCACCCACATAGCCC	31	3456	3471	291

908203	116729	116744	CAAGACCCCCCGCCAC	64	3473	3488	292

908204	116732	116747	CCCCAAGACCCCCCGC	59	3476	3491	293

908205	116738	116753	AGAGGCCCCCAAGACC	86	3482	3497	294

908206	116772	116787	GGCCCACCCATCAGGG	110	3516	3531	295

908207	116774	116789	TGGGCCCACCCATCAG	127	3518	3533	296

908208	116789	116804	AGAGAGAGGCCGCCCT	37	3533	3548	297

908209	116791	116806	TCAGAGAGAGGCCGCC	70	3535	3550	298

908210	116808	116823	GAGTGGGTGAGGTCTC	77	3552	3567	299

908211	116815	116830	GAGCGAGGAGTGGGTG	59	3559	3574	300

908212	116819	116834	AACTGAGCGAGGAGTG	55	3563	3578	301

908213	116822	116837	TCAAACTGAGCGAGGA	5	3566	3581	302

908214	116826	116841	GTGGTCAAACTGAGCG	15	3570	3585	303

908215	116832	116847	CTTACAGTGGTCAAAC	10	3576	3591	304

908216	116843	116858	GAGTGCAGGCACTTAC	48	3587	3602	305

908217	116849	116864	AATACAGAGTGCAGGC	4	3593	3608	306

908221	82462	82477	CCACGAGATCAACACA	20	N/A	N/A	307

908222	82464	82479	GACCACGAGATCAACA	88	N/A	N/A	308

908223	82465	82480	AGACCACGAGATCAAC	82	N/A	N/A	309

908224	82468	82483	CTGAGACCACGAGATC	34	N/A	N/A	310

908225	82471	82486	GCTCTGAGACCACGAG	59	N/A	N/A	311

908226	82482	82497	GACCGTGGCCAGCTCT	52	N/A	N/A	312

908227	82484	82499	ATGACCGTGGCCAGCT	54	N/A	N/A	313

908228	82485	82500	TATGACCGTGGCCAGC	31	N/A	N/A	314

908229	82487	82502	TTTATGACCGTGGCCA	64	N/A	N/A	315

908230	82488	82503	TTTTATGACCGTGGCC	102	N/A	N/A	316

908231	82498	82513	CGGCACACTTTTTTAT	37	N/A	N/A	317

908232	82502	82517	TTCTCGGCACACTTTT	4	N/A	N/A	318

908236	N/A	N/A	CCCCGCCGCGGGCACG	103	3506	3521	319

908237	4387	4402	CCAAAGGAGGGCCCCC	88	N/A	N/A	320

908238	4857	4872	CGCCAGCCTAGTGGCC	142	N/A	N/A	321

908239	5585	5600	TGGGACTACCACCCTA	129	N/A	N/A	322

908240	6095	6110	CTCCATACTGGCCGGC	91	N/A	N/A	323

908241	6449	6464	CTTAACGAGGCCAGCC	56	N/A	N/A	324

908242	7057	7072	ACCCATCTAGGCCCCA	23	N/A	N/A	325

908243	7335	7350	TGGTGCACCGCAGACA	61	N/A	N/A	326

908244	8045	8060	TCAGGAGGCGAGATTG	93	N/A	N/A	327

908245	8318	8333	CGGGAAAACCCCGCCT	123	N/A	N/A	328

908246	8813	8828	AGCCACTAAGTGGCCC	130	N/A	N/A	329

908247	9601	9616	GCTAAACTGGCCTCCG	78	N/A	N/A	330

908248	9989	10004	GCCAACGGGCATCCGA	104	N/A	N/A	331

908249	10650	10665	GTCCACTCCCAGTAGG	77	N/A	N/A	332

908250	11631	11646	GGTGATCCGATGCTGG	34	N/A	N/A	333

908251	12300	12315	GGCCATCTTCCGCCCT	102	N/A	N/A	334

908252	12662	12677	CTAACCTAGGCCGGGT	70	N/A	N/A	335

908253	12991	13006	AGACGCAGGGCCACAT	66	N/A	N/A	336

908254	13381	13396	GGCTCTAGAGGGCCCA	110	N/A	N/A	337

908255	13954	13969	CCGCATACAGCCTTTT	12	N/A	N/A	338

908256	14620	14635	GCATTCCACGGGCAGC	34	N/A	N/A	339

908257	15396	15411	CACCGCTACCCTAGCT	63	N/A	N/A	340

908258	15792	15807	GGGCACCCCGCAGGTT	102	N/A	N/A	341

908259	16235	16250	TCTAGAAGGACCCTGT	64	N/A	N/A	342

908260	16810	16825	AGCCGGGCTTCCGTCT	84	N/A	N/A	343

908261	17238	17253	TTTAAGGGCGCGGTGG	99	N/A	N/A	344

908262	17624	17639	TGCACTTCCGGACAGG	82	N/A	N/A	345

908263	18173	18188	CACCAGCCCAGGCGCG	117	N/A	N/A	346

908264	18617	18632	AGCTAGGTTAATTTCT	64	N/A	N/A	347

908265	19070	19085	CAACATCCCGCCAGGC	80	N/A	N/A	348

908266	19407	19422	AGCAATCTAGCCTCCC	39	N/A	N/A	349

908267	19819	19834	TGCAATGTCCAGTCCT	11	N/A	N/A	350

908268	20435	20450	ACCTAGCAAGCCTACC	64	N/A	N/A	351

908269	21415	21430	GCTACAGCAATATAGA	67	N/A	N/A	352

908270	21839	21854	TCTTAACAGGCTCCCT	63	N/A	N/A	353

908271	22199	22214	CCTCAGGGACGACTTC	87	N/A	N/A	354

908272	22636	22651	TATACGGGCATGTTGA	55	N/A	N/A	355

908273	22916	22931	GTGCATCCCGAGAGGG	89	N/A	N/A	356

908274	23292	23307	ATGAAAGTCTCCACTT	61	N/A	N/A	357

908275	23543	23558	AACCGCTGGGATCCCC	71	N/A	N/A	358

908276	24229	24244	GTACAGGAGTTCCCCA	90	N/A	N/A	359

908277	25038	25053	AAATACACCCTAGATG	83	N/A	N/A	360

908278	25670	25685	GAAGATTTTGGCCGGG	85	N/A	N/A	361

908279	25997	26012	CTCCAGGACACACCGT	42	N/A	N/A	362

908280	27088	27103	GGGAGTAGGCTCGACC	51	N/A	N/A	363

908281	27400	27415	CCCTATTCCACAGGAC	85	N/A	N/A	364

908282	27746	27761	GCTAAGAGCAGCCCTC	80	N/A	N/A	365

908283	28198	28213	GGCATCACGGCCGGGC	76	N/A	N/A	366

908284	29125	29140	TGTCACCCCAGCCGAA	67	N/A	N/A	367

908285	29568	29583	AGCCGGGCACCACCCA	67	N/A	N/A	368

908286	30330	30345	GGCAAGGACAGCCGAC	94	N/A	N/A	369

908287	30334	30349	GTTTGGCAAGGACAGC	40	N/A	N/A	370

908288	30337	30352	GTGGTTTGGCAAGGAC	14	N/A	N/A	371

908289	30340	30355	GCAGTGGTTTGGCAAG	46	N/A	N/A	372

908290	30342	30357	ATGCAGTGGTTTGGCA	86	N/A	N/A	373

908291	30401	30416	TCAACACCTTGAGGGC	34	N/A	N/A	374

908292	30498	30513	TGCCTAGGCGATCCTC	88	N/A	N/A	375

908293	30928	30943	GACATTGCTGTCGCAG	78	N/A	N/A	376

908294	31347	31362	TCCCAAGTGCCCCCCA	90	N/A	N/A	377

908295	31703	31718	AGCCGGCACGGTTGCT	125	N/A	N/A	378

908296	32187	32202	GAGAACATTCGCCCCA	27	N/A	N/A	379

908297	32386	32401	TGACCTGTGGCAATTC	26	N/A	N/A	380

908298	32388	32403	GGTGACCTGTGGCAAT	44	N/A	N/A	381

908299	32689	32704	TCCCGTATCGGCCCCT	85	N/A	N/A	382

908300	33343	33358	GTCCAGCCGCGAGTCC	59	N/A	N/A	383

908301	33635	33650	GGCAGACCCGCAGTCA	73	N/A	N/A	384

908302	34108	34123	GGGCAAGCCACCCCTC	93	N/A	N/A	385

908303	34623	34638	AAGTCGAGCTCCGCCC	63	N/A	N/A	386

908304	35031	35046	TGGTCCTAGCCCGTCC	78	N/A	N/A	387

908305	35549	35564	GGGTACAGGCAAACCT	88	N/A	N/A	388

908306	36028	36043	CCATACCCCCATCTGA	102	N/A	N/A	389

908307	36712	36727	GCGCAGAGGGCCTCGC	123	N/A	N/A	390

908308	37097	37112	CACTACAGACATCCCT	64	N/A	N/A	391

908309	37538	37553	AGCCAGGGCCCGGCTT	93	N/A	N/A	392

908310	38044	38059	CTCCAACTGTAGGGCT	87	N/A	N/A	393

908311	39735	39750	GCGGATGGTGGCATTC	90	N/A	N/A	394

908312	40399	40414	CAGCAGCGGCGGTGAC	84	N/A	N/A	395

908313	41143	41158	CTCTTAGCATGGCCCC	73	N/A	N/A	396

908314	41620	41635	GGCCTGGGATGCTACC	129	N/A	N/A	397

908315	41957	41972	TTAAACCCCCCTCTGG	104	N/A	N/A	398

908316	42239	42254	GCCCAAGCCCTCGGCG	147	N/A	N/A	399

908317	42806	42821	ATGCACGCGCTACCAT	106	N/A	N/A	400

908318	43841	43856	ACATATCCTGGCCGGG	112	N/A	N/A	401

908319	44266	44281	CTGGATGTGGGAACGG	94	N/A	N/A	402

908320	44817	44832	CCTCAGGCCGCCCCAT	57	N/A	N/A	403

908321	45179	45194	GGCCATGGAGGGTCAC	111	N/A	N/A	404

908322	45544	45559	GGGACAACTGGCGACA	39	N/A	N/A	405

908323	45952	45967	TGGTAGAGGGCTGGTG	78	N/A	N/A	406

908324	46827	46842	GCCCATCGGCTGCTGT	153	N/A	N/A	407

908325	47604	47619	GGGTAGGCCTGCGCTC	119	N/A	N/A	408

908326	48030	48045	TGCCATGATCAGGCGG	109	N/A	N/A	409

908327	48530	48545	ACCAAGTGGGCACCTA	92	N/A	N/A	410

908328	49088	49103	CCACAAGGCCCCGCCG	103	N/A	N/A	411

908329	49883	49898	GTGACCAGGTGCGGGT	84	N/A	N/A	412

908330	50310	50325	CTGGGCAGAGCCGATC	95	N/A	N/A	413

908331	50840	50855	GACGAGGCTGGCCCTG	36	N/A	N/A	414

908332	51504	51519	CCCCACTGTGCTAAAC	91	N/A	N/A	415

908333	51765	51780	AGTCAGCCGTCCTCGC	58	N/A	N/A	416

908334	52587	52602	CACCAGGCCGAGAGCA	111	N/A	N/A	417

908335	53006	53021	GCCCAGGCACCGACCA	92	N/A	N/A	418

908336	53680	53695	CCCCGGAACCTCAGGC	85	N/A	N/A	419

908337	54271	54286	CAGCACGCTGTGTGCA	65	N/A	N/A	420

908338	54667	54682	CACTCGGCCGCTCTTC	80	N/A	N/A	421

908339	55325	55340	GCGCACTGCCCTGTCG	77	N/A	N/A	422

908340	56452	56467	CAAGAGGGCCCCCTAC	110	N/A	N/A	423

908341	56855	56870	TATGAATCCCCCTCCC	99	N/A	N/A	424

908342	57413	57428	GCGCATGGACTGCGGG	150	N/A	N/A	425

908343	57695	57710	TGGAAGTACACAGGCT	40	N/A	N/A	426

908344	58949	58964	GTCTATGGCCCCGGGC	102	N/A	N/A	427

908345	59713	59728	GCATAGGCCAACGCAG	88	N/A	N/A	428

908346	60635	60650	AAACAGGCCCGCAGCC	95	N/A	N/A	429

908347	61004	61019	CCTACAGCGGCCATGG	115	N/A	N/A	430

908348	61545	61560	GTGCTAACCGTGTGTG	85	N/A	N/A	431

908349	62025	62040	GTCCAGTTGGCCCTGA	84	N/A	N/A	432

908350	62964	62979	AACCACCGAAGCTGGG	125	N/A	N/A	433

908351	63324	63339	GGAAGATGGCCGGGTG	111	N/A	N/A	434

908352	63675	63690	GCTACAATGCCTAGGA	85	N/A	N/A	435

908353	64393	64408	GGTCAACGCAGGCTCA	48	N/A	N/A	436

908354	64657	64672	AGCCAGGAGCAATCGC	78	N/A	N/A	437

908355	65114	65129	GGCAACCAAGGTCTGA	15	N/A	N/A	438

908356	65639	65654	CGCCTAGCCTGGGTTC	67	N/A	N/A	439

908357	66042	66057	CACCATCACACACGGA	44	N/A	N/A	440

908358	66433	66448	GGGAGAGGTGCCGGTC	99	N/A	N/A	441

908359	66885	66900	CACCGAGAAGGAGGTA	99	N/A	N/A	442

908360	67650	67665	AAAATCTCGGCCGGGA	101	N/A	N/A	443

908361	67903	67918	GCCAAGTGGACCTGGC	84	N/A	N/A	444

908362	68192	68207	GCCTGAAACAAGTGCC	75	N/A	N/A	445

908363	68628	68643	CCCCAGGCAATGCATA	125	N/A	N/A	446

908364	68993	69008	AGAACGATAGGCCAGG	52	N/A	N/A	447

908365	69310	69325	TCTTTTCCCGGCCCCC	48	N/A	N/A	448

908366	71364	71379	CGGCACGGAGCTGCCC	126	N/A	N/A	449

908367	71557	71572	GGTCAGTTGCTGCAGG	40	N/A	N/A	450

908368	72040	72055	GGCCGGCCAATGCTCA	118	N/A	N/A	451

908369	73315	73330	TTGCACTGGCCGGGCT	75	N/A	N/A	452

908370	73937	73952	CCCTTAGCCCACGACC	92	N/A	N/A	453

908371	74402	74417	CTGCCAGTTTGTCCCC	25	N/A	N/A	454

908372	74408	74423	TGTAATCTGCCAGTTT	53	N/A	N/A	455

908373	74410	74425	CTTGTAATCTGCCAGT	55	N/A	N/A	456

908374	74415	74430	GGCACCTTGTAATCTG	96	N/A	N/A	457

908375	74908	74923	TGTGATGGGCATCCCC	56	N/A	N/A	458

908376	75761	75776	ATACATGAGGTCCGGG	106	N/A	N/A	459

908377	76583	76598	GCCCAACCCACGCTGC	79	N/A	N/A	460

908378	77237	77252	CCCCAACTGGGAGAGG	107	N/A	N/A	461

908379	77517	77532	TAGAACGGGCCTGGCC	93	N/A	N/A	462

908380	78331	78346	TCCATAGGTTCCCGCC	89	N/A	N/A	463

908381	78849	78864	GGGCACTACCTTCACC	138	N/A	N/A	464

908382	79488	79503	TCCCACGGATTATAGG	140	N/A	N/A	465

908383	80002	80017	GCCCAGGAAGGTCGAA	105	N/A	N/A	466

908384	80333	80348	ATGCATGGAGGCACCC	93	N/A	N/A	467

908385	80626	80641	TGCAATGGACGCAGGA	37	N/A	N/A	468

908386	80968	80983	GGCCGGGTCCCCAACA	113	N/A	N/A	469

908387	81643	81658	GGCCATTCCCGGAGCT	114	N/A	N/A	470

908388	82021	82036	CCTTTTCTAATCGAAG	116	N/A	N/A	471

908389	82046	82061	ACCTGTTTGGACTGTT	5	N/A	N/A	472

908390	82049	82064	CCAACCTGTTTGGACT	74	N/A	N/A	473

908391	82051	82066	TTCCAACCTGTTTGGA	54	N/A	N/A	474

908392	82055	82070	CGTTTTCCAACCTGTT	3	N/A	N/A	475

908393	82069	82084	GGCTCTAGGACGAGCG	55	N/A	N/A	476

908394	82072	82087	CCTGGCTCTAGGACGA	41	N/A	N/A	477

908395	82083	82098	TGTCCATGACACCTGG	66	N/A	N/A	478

908396	82086	82101	TCATGTCCATGACACC	8	N/A	N/A	479

908397	82088	82103	ATTCATGTCCATGACA	25	N/A	N/A	480

908398	82704	82719	CCCCACGGGCAGCCAC	127	N/A	N/A	481

908399	83290	83305	GGGCACCGTGGACCGG	112	N/A	N/A	482

908400	83646	83661	CATATGCAGCTGTCAG	27	N/A	N/A	483
	83734	83749			N/A	N/A
	83822	83837			N/A	N/A

908401	83647	83662	CCATATGCAGCTGTCA	2	N/A	N/A	484
	83735	83750			N/A	N/A
	83823	83838			N/A	N/A

908402	83649	83664	TGCCATATGCAGCTGT	27	N/A	N/A	485
	83737	83752			N/A	N/A
	83825	83840			N/A	N/A

908403	83650	83665	TTGCCATATGCAGCTG	18	N/A	N/A	486
	83738	83753			N/A	N/A
	83826	83841			N/A	N/A

908404	83651	83666	TTTGCCATATGCAGCT	17	N/A	N/A	487
	83739	83754			N/A	N/A
	83827	83842			N/A	N/A

908405	83658	83673	GGGTTACTTTGCCATA	7	N/A	N/A	488
	83746	83761			N/A	N/A
	83834	83849			N/A	N/A

908406	83659	83674	AGGGTTACTTTGCCAT	74	N/A	N/A	489
	83747	83762			N/A	N/A
	83835	83850			N/A	N/A

908407	83660	83675	AAGGGTTACTTTGCCA	7	N/A	N/A	490
	83748	83763			N/A	N/A
	83836	83851			N/A	N/A

908408	83661	83676	TAAGGGTTACTTTGCC	2	N/A	N/A	491
	83749	83764			N/A	N/A
	83837	83852			N/A	N/A

908409	83662	83677	GTAAGGGTTACTTTGC	8	N/A	N/A	492
	83750	83765			N/A	N/A
	83838	83853			N/A	N/A

908410	83663	83678	GGTAAGGGTTACTTTG	12	N/A	N/A	493
	83751	83766			N/A	N/A
	83839	83854			N/A	N/A

908411	83664	83679	AGGTAAGGGTTACTTT	17	N/A	N/A	494
	83752	83767			N/A	N/A
	83840	83855			N/A	N/A

908412	83665	83680	GAGGTAAGGGTTACTT	12	N/A	N/A	495
	83753	83768			N/A	N/A
	83841	83856			N/A	N/A

908413	83666	83681	TGAGGTAAGGGTTACT	3	N/A	N/A	496
	83754	83769			N/A	N/A
	83842	83857			N/A	N/A

908414	83667	83682	TTGAGGTAAGGGTTAC	5	N/A	N/A	497
	83755	83770			N/A	N/A
	83843	83858			N/A	N/A

908415	83669	83684	AACCCTTACCTCAATC	3	N/A	N/A	498
	83757	83772			N/A	N/A
	83845	83860			N/A	N/A

908416	83670	83685	TGATTGAGGTAAGGGT	4	N/A	N/A	499
	83758	83773			N/A	N/A
	83846	83861			N/A	N/A

908417	83671	83686	TTGATTGAGGTAAGGG	2	N/A	N/A	500
	83759	83774			N/A	N/A
	83847	83862			N/A	N/A

908418	83672	83687	CTTGATTGAGGTAAGG	8	N/A	N/A	501
	83760	83775			N/A	N/A
	83848	83863			N/A	N/A

908419	83673	83688	TCTTGATTGAGGTAAG	5	N/A	N/A	502
	83761	83776			N/A	N/A
	83849	83864			N/A	N/A

908420	83674	83689	ATCTTGATTGAGGTAA	2	N/A	N/A	503
	83762	83777			N/A	N/A
	83850	83865			N/A	N/A

908421	83684	83699	ACCGCTCTGTATCTTG	3	N/A	N/A	504
	83772	83787			N/A	N/A
	83860	83875			N/A	N/A

908422	83685	83700	AACCGCTCTGTATCTT	3	N/A	N/A	505
	83773	83788			N/A	N/A
	83861	83876			N/A	N/A

908423	83686	83701	AGATACAGAGCGGTTC	28	N/A	N/A	506
	83774	83789			N/A	N/A
	83862	83877			N/A	N/A

908424	83687	83702	GGAACCGCTCTGTATC	8	N/A	N/A	507
	83775	83790			N/A	N/A
	83863	83878			N/A	N/A

908425	83688	83703	CGGAACCGCTCTGTAT	13	N/A	N/A	508
	83776	83791			N/A	N/A
	83864	83879			N/A	N/A

908426	83689	83704	ACGGAACCGCTCTGTA	47	N/A	N/A	509
	83777	83792			N/A	N/A
	83865	83880			N/A	N/A

908427	83690	83705	GACGGAACCGCTCTGT	21	N/A	N/A	510
	83778	83793			N/A	N/A
	83866	83881			N/A	N/A

908428	83691	83706	TGACGGAACCGCTCTG	33	N/A	N/A	511
	83779	83794			N/A	N/A
	83867	83882			N/A	N/A

908429	83692	83707	GTGACGGAACCGCTCT	44	N/A	N/A	512
	83780	83795			N/A	N/A
	83868	83883			N/A	N/A

908430	83693	83708	GGTGACGGAACCGCTC	27	N/A	N/A	513
	83781	83796			N/A	N/A
	83869	83884			N/A	N/A

908431	83694	83709	AGCGGTTCCGTCACCT	67	N/A	N/A	514
	83782	83797			N/A	N/A
	83870	83885			N/A	N/A

908432	83695	83710	GAGGTGACGGAACCGC	69	N/A	N/A	515
	83783	83798			N/A	N/A
	83871	83886			N/A	N/A

908433	83696	83711	GGAGGTGACGGAACCG	62	N/A	N/A	516
	83784	83799			N/A	N/A
	83872	83887			N/A	N/A

908434	83697	83712	AGGAGGTGACGGAACC	6	N/A	N/A	517
	83785	83800			N/A	N/A
	83873	83888			N/A	N/A

908435	83698	83713	TAGGAGGTGACGGAAC	16	N/A	N/A	518
	83786	83801			N/A	N/A
	83874	83889			N/A	N/A

908436	83699	83714	TTAGGAGGTGACGGAA	14	N/A	N/A	519
	83787	83802			N/A	N/A
	83875	83890			N/A	N/A

908437	83709	83724	GACAGGGATTTTAGGA	29	N/A	N/A	520
	83885	83900			N/A	N/A

908438	83710	83725	GGACAGGGATTTTAGG	49	N/A	N/A	521
	83886	83901			N/A	N/A

908439	83719	83734	CCTGTCCCTATCATGC	3	N/A	N/A	522
	83807	83822			N/A	N/A

908440	83720	83735	AGCATGATAGGGACAG	4	N/A	N/A	523
	83808	83823			N/A	N/A

908441	83721	83736	CAGCATGATAGGGACA	2	N/A	N/A	524
	83809	83824			N/A	N/A

908442	83722	83737	TCAGCATGATAGGGAC	4	N/A	N/A	525
	83810	83825			N/A	N/A

908443	83723	83738	GTCAGCATGATAGGGA	4	N/A	N/A	526
	83811	83826			N/A	N/A

908444	83724	83739	TGTCAGCATGATAGGG	4	N/A	N/A	527
	83812	83827			N/A	N/A

908445	83725	83740	CTGTCAGCATGATAGG	44	N/A	N/A	528
	83813	83828			N/A	N/A

908446	83726	83741	GCTGTCAGCATGATAG	70	N/A	N/A	529
	83814	83829			N/A	N/A

908447	83727	83742	TATCATGCTGACAGCT	55	N/A	N/A	530
	83815	83830			N/A	N/A

908448	83910	83925	GAGGTTGACTACAAAG	12	N/A	N/A	531

908449	83913	83928	TGAGAGGTTGACTACA	6	N/A	N/A	532

908450	83915	83930	TTTGAGAGGTTGACTA	17	N/A	N/A	533

908451	83921	83936	TGGGTTTTTGAGAGGT	9	N/A	N/A	534

908452	84177	84192	GGCGATGGCACGACCA	71	N/A	N/A	535

908453	84938	84953	GTATAAGGACGGCCCA	45	N/A	N/A	536

908454	85500	85515	TCAGATCCGGCCTGGC	191	N/A	N/A	537

908455	86165	86180	GTGGTGTGGCGTGCGC	75	N/A	N/A	538

908456	86583	86598	GAACTTACTGTCTCCC	14	N/A	N/A	539

908457	87056	87071	AGGCAGGGCCGCTGTC	100	N/A	N/A	540

908458	87386	87401	AGGATTGGTGCCGCCC	36	N/A	N/A	541

908459	87906	87921	AATCAGCGGGTGATGC	118	N/A	N/A	542

908460	88179	88194	GCCCAGTGGCCGCCCT	70	N/A	N/A	543

908461	88831	88846	CTACAGCTGGTGCGGT	40	N/A	N/A	544

908462	89400	89415	CGCCAGGCTTGGGCCC	100	N/A	N/A	545

908463	89711	89726	GCCCAGATAAGAAGCC	38	N/A	N/A	546

908464	90096	90111	GGCTAGTTCATGCCGG	63	N/A	N/A	547

908465	90743	90758	ACGGCCTGGGCACACG	52	N/A	N/A	548

908466	90747	90762	GGAAACGGCCTGGGCA	64	N/A	N/A	549

908467	90750	90765	AAGGGAAACGGCCTGG	76	N/A	N/A	550

908468	90753	90768	CGCAAGGGAAACGGCC	108	N/A	N/A	551

908469	90755	90770	TGCGCAAGGGAAACGG	87	N/A	N/A	552

908470	90757	90772	GCTGCGCAAGGGAAAC	124	N/A	N/A	553

908471	90761	90776	CCCTTGCGCAGCTCTG	18	N/A	N/A	554

908472	90765	90780	CACACAGAGCTGCGCA	57	N/A	N/A	555

908473	90778	90793	GAAGCCCCCTGCACAC	37	N/A	N/A	556

908474	90781	90796	AGAGAAGCCCCCTGCA	72	N/A	N/A	557

908475	90809	90824	CCAAGTGAGCAGCCAC	9	N/A	N/A	558

908476	90811	90826	GACCAAGTGAGCAGCC	27	N/A	N/A	559

908477	90813	90828	GGGACCAAGTGAGCAG	63	N/A	N/A	560

908478	91157	91172	CACTAGGCCCTGCCAG	128	N/A	N/A	561

908479	91836	91851	GAGCGGGGGTCTGTGA	58	N/A	N/A	562

908480	92802	92817	CAGCACTTTGTCCGGG	82	N/A	N/A	563

908481	93315	93330	ACCTACGGCAAGCGCA	31	N/A	N/A	564

908482	93568	93583	TACCAGGATCTCCCCC	19	N/A	N/A	565

908483	94077	94092	GCTAGAGGAGCGGCAC	74	N/A	N/A	566

908484	94531	94546	GCCCATCTCACGCCCC	71	N/A	N/A	567

908485	95058	95073	CCAGAGGCCACGGCCA	94	N/A	N/A	568

908486	95635	95650	CCGCACACCCCTCGGT	111	N/A	N/A	569

908487	96522	96537	CAGGCCGCCCTGTGCG	93	N/A	N/A	570

908488	97096	97111	CCCAAGCATTTCCAGC	37	N/A	N/A	571

908489	97109	97124	AGTTCCACCCTGTCCC	23	N/A	N/A	572

908490	97144	97159	TAGAGACACCTCCAGC	47	N/A	N/A	573

908491	97147	97162	CAATAGAGACACCTCC	44	N/A	N/A	574

908492	97150	97165	CCGCAATAGAGACACC	43	N/A	N/A	575

908493	97152	97167	GACCGCAATAGAGACA	64	N/A	N/A	576

908494	97154	97169	GGGACCGCAATAGAGA	94	N/A	N/A	577

908495	97157	97172	CTATTGCGGTCCCTGG	31	N/A	N/A	578

908496	97159	97174	AGCCAGGGACCGCAAT	85	N/A	N/A	579

908497	97426	97441	GCCCACCCGCTCTTCC	80	N/A	N/A	580

908498	98041	98056	TGGAACCCCGGTGCCC	119	N/A	N/A	581

908499	98971	98986	CTTCACCGTTTGGCCA	45	N/A	N/A	582

908500	100149	100164	CGCGGAGCCAGGCCCT	114	N/A	N/A	583

908501	101337	101352	CCCCAGAGTCCCGCAG	72	N/A	N/A	584

908502	101845	101860	ACACGAAGGGCCCCTC	52	N/A	N/A	585

908503	102223	102238	GCAGACTGATAGGGCC	103	N/A	N/A	586

908504	102495	102510	GGATAGCCGGCCCCGC	102	N/A	N/A	587

908505	102955	102970	CGCGGCCGGAAGCTTC	93	N/A	N/A	588

908506	103211	103226	CAGAGCATAGGAGAGG	7	N/A	N/A	589
	103249	103264			N/A	N/A

908507	103213	103228	AGCAGAGCATAGGAGA	12	N/A	N/A	590
	103251	103266			N/A	N/A

908508	103216	103231	GGGAGCAGAGCATAGG	16	N/A	N/A	591
	103254	103269			N/A	N/A

908509	104082	104097	GCCAGCCACGGCTGTC	81	N/A	N/A	592

908510	104495	104510	GCTGACGGCACGCACA	65	N/A	N/A	593

908511	105108	105123	TGGGGATGGCTCGGGG	79	N/A	N/A	594

908512	105526	105541	CACCAGGGCTCCGCCT	107	N/A	N/A	595

908513	106268	106283	GGCACACGGCTCAGGG	93	N/A	N/A	596

908514	106974	106989	CAGCAAGTGGCTGCCC	107	N/A	N/A	597

908515	107645	107660	TGGGAGTGGTACTGCA	70	N/A	N/A	598

908516	108733	108748	TGCCATGGGTCTGGCA	86	N/A	N/A	599

908517	108893	108908	GCTCCTCACCTGGTCC	92	N/A	N/A	600

908518	108899	108914	AGGACGGCTCCTCACC	111	N/A	N/A	601

908519	108902	108917	GAGGAGCCGTCCTGCG	85	N/A	N/A	602

908520	108906	108921	GCTGCGCAGGACGGCT	104	N/A	N/A	603

908521	109226	109241	GGGCGAACCACCCACC	78	N/A	N/A	604

908522	109577	109592	GCCCAAAGGATCTGCC	88	N/A	N/A	605

908523	109968	109983	CCCTATTGTCCCACCA	43	N/A	N/A	606

908524	110612	110627	GTGCACTCTAGGCTTA	100	N/A	N/A	607

908525	111378	111393	GGACACAGCGGCTGCC	78	N/A	N/A	608

908526	111925	111940	CCGCAGGTTTCTGTGG	122	N/A	N/A	609

908527	112198	112213	ACCGATAGGATCGGAG	95	N/A	N/A	610

908528	112578	112593	GGCCGTCACAACTTTT	116	N/A	N/A	611

908529	112910	112925	CTGGAACCTCCTACTG	87	N/A	N/A	612

908530	114292	114307	GGCCGAATGCTTGAGG	97	N/A	N/A	613

908531	114921	114936	TCACAGGGCCCCCTCA	68	N/A	N/A	614

908532	115371	115386	GGCCTTACCTGGGCAC	101	N/A	N/A	615

908533	115837	115852	CCGCAAGCCACCAGGG	87	N/A	N/A	616

*Oligos with an asterisk target an amplicon region.

Each modified oligonucleotide listed in the table below is 100% complementary to human DNM2 nucleic acid sequence GENBANK Number NM_001005361.2 (designated herein as SEQ ID NO:3). “Start Site” indicates the 5′-most nucleoside of SEQ ID No: 3 to which the oligonucleotide is complementary. “Stop Site” indicates the 3′-most nucleoside of SEQ ID No: 3 to which the oligonucleotide is complementary. ‘N/A’ indicates that the modified oligonucleotide is not 100% complementary to the particular nucleic acid. As shown below, modified oligonucleotides complementary to DNM2 reduced human DNM2 mRNA expression.

TABLE 2

DNM2 mRNA Expression

	SEQ	SEQ
	ID: 3	ID: 3			SEQ
Compound	Start	Stop		Dynamin2	ID
Number	Site	Site	Sequence	(%Control)	NO

695118	1728	1743	ATCTCCCCCTGATTGG	60	617

908218	1732	1747	CAGGATCTCCCCCTGA	114	618

908219	1379	1394	GCCCCGTCCTGACTCC	61	619

908220	1381	1396	GAGCCCCGTCCTGACT	92	620

908233	1515	1530	CTGAGCTTCTCGGCAC	70	621

908234	1517	1532	AACTGAGCTTCTCGGC	85	622

908235	1519	1534	GGAACTGAGCTTCTCG	33	623

Example 2: Effect of Modified Oligonucleotides Complementary to Dynamin 2 In Vitro

Modified oligonucleotides complementary to dynamin (DNM2) nucleic acid were designed and tested for their effect on dynamin 2 mRNA in vitro. The modified oligonucleotides were tested in a series of experiments that had similar culture conditions.
A431 cells cultured at a density of 10,000 cells per well were treated with 2,000 nM of modified oligonucleotide via free uptake or no oligonucleotide for untreated controls. After approximately 24 hours, RNA was isolated from the cells and DNM2 mRNA levels were measured by quantitative real-time PCR as described in Example 1.
The modified oligonucleotides in the tables below each have a 3-10-3 cEt gapmer motif, wherein the central gap segment contains ten 2′-deoxynucleosides and is flanked by wing segments on the 3′ and 5′ ends, each containing three cEt nucleosides. All cytosine residues throughout each modified oligonucleotide are 5-methyl cytosines. All internucleoside linkages are phosphorothioate internucleoside linkages.
Each modified oligonucleotide listed in the tables below is 100% complementary to human DNM2 nucleic acid sequence GENBANK Number NC_000019.10, truncated from 10715001 to 1083500 (SEQ ID NO: 1) and/or GENBANK Number NM_004945.3 (SEQ ID NO:2). As shown below, modified oligonucleotides complementary to human DNM2 inhibited human DNM2 mRNA expression.

TABLE 3

DNM2 mRNA Expression

					SEQ	SEQ
	SEQ ID:	SEQ ID:			ID: 2	ID 2:	SEQ
Compound	1 Start	1 Stop		Dunamin2	Start	Stop	ID
Number	Site	Site	Sequence	(%Control)	Site	Site	NO

694792	3060	3075	CGACACCCGACCCGAG	26	N/A	N/A	624

694812	3317	3332	GTCCAGGTGGCAGCTC	37	265	280	625

694826	57522	57537	CCGGACTTCATCAAAG	82	469	484	626

694999	115263	115278	GTCCAGGCCGGGATGG	92	2606	N/A	627

695015	116005	116020	GCGGATAATGGTGGGC	12	2749	N/A	628

695016	116011	116026	GGCTGGGCGGATAATG	76	2755	N/A	629

695170	82446	82461	CTTCAAACTCGGCTCT	9	N/A	N/A	630

695171	82451	82466	ACACACTTCAAACTCG	7	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	12	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	9	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	8	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	11	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	7	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	121	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	14	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	8	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	6	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	31	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	8	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	6	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	10	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	8	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	12	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	11	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	11	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	5	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	12	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	7	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	120	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	6	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	8	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	9	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	7	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	10	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	9	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	9	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	9	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	10	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	7	N/A	N/A	13

948484	82379	82394	GTGAAGAGCCCCGTCC	42	N/A	N/A	631

948485	82380	82395	GGTGAAGAGCCCCGTC	103	N/A	N/A	632

948486	82443	82458	CAAACTCGGCTCTTTG	99	N/A	N/A	633

948487	82444	82459	TCAAACTCGGCTCTTT	15	N/A	N/A	634

948488	82445	82460	TTCAAACTCGGCTCTT	5	N/A	N/A	635

948489	82448	82463	CACTTCAAACTCGGCT	3	N/A	N/A	636

948490	82449	82464	ACACTTCAAACTCGGC	1	N/A	N/A	637

948491	82450	82465	CACACTTCAAACTCGG	25	N/A	N/A	638

948492	82452	82467	AACACACTTCAAACTC	11	N/A	N/A	639

948493	82453	82468	CAACACACTTCAAACT	51	N/A	N/A	640

948494	82454	82469	TCAACACACTTCAAAC	30	N/A	N/A	641

948495	82458	82473	GAGATCAACACACTTC	19	N/A	N/A	642

948496	82460	82475	ACGAGATCAACACACT	5	N/A	N/A	643

948497	82461	82476	CACGAGATCAACACAC	28	N/A	N/A	644

948498	82463	82478	ACCACGAGATCAACAC	50	N/A	N/A	645

948499	82467	82482	TGAGACCACGAGATCA	55	N/A	N/A	646

948500	82469	82484	TCTGAGACCACGAGAT	107	N/A	N/A	647

948501	82470	82485	CTCTGAGACCACGAGA	119	N/A	N/A	648

948502	82490	82505	TTTTTTATGACCGTGG	18	N/A	N/A	649

948503	82496	82511	GCACACTTTTTTATGA	17	N/A	N/A	650

948504	82497	82512	GGCACACTTTTTTATG	54	N/A	N/A	651

948505	82499	82514	TCGGCACACTTTTTTA	10	N/A	N/A	652

948506	82500	82515	CTCGGCACACTTTTTT	3	N/A	N/A	653

948507	82501	82516	TCTCGGCACACTTTTT	3	N/A	N/A	654

948508	82503	82518	CTTCTCGGCACACTTT	3	N/A	N/A	655

948514	3056	3071	ACCCGACCCGAGCGAC	81	4	19	656

948515	3067	3082	TCTCAGGCGACACCCG	88	15	30	657

948516	3092	3107	CCTCACGGTCGCCGCC	56	40	55	658

948517	3094	3109	GGCCTCACGGTCGCCG	98	42	57	659

948518	3103	3118	TCCCGGCTCGGCCTCA	48	51	66	660

948519	3139	3154	GCTCCCCGCCCGCCCG	71	87	102	661

948520	3141	3156	TTGCTCCCCGCCCGCC	23	89	104	662

948521	3142	3157	GTTGCTCCCCGCCCGC	10	90	105	663

948522	3143	3158	CGTTGCTCCCCGCCCG	50	91	106	664

948523	3145	3160	GCCGTTGCTCCCCGCC	53	93	108	665

948524	3147	3162	TAGCCGTTGCTCCCCG	17	95	110	666

948525	3148	3163	GTAGCCGTTGCTCCCC	14	96	111	667

948526	3150	3165	CTGTAGCCGTTGCTCC	10	98	113	668

948527	3151	3166	TCTGTAGCCGTTGCTC	9	99	114	669

948528	3152	3167	GTCTGTAGCCGTTGCT	13	100	115	670

948529	3168	3183	GACCTGGCCCCGCGGC	76	116	131	671

948530	3169	3184	CGACCTGGCCCCGCGG	83	117	132	672

948531	3170	3185	ACGACCTGGCCCCGCG	36	118	133	673

948532	3172	3187	CAACGACCTGGCCCCG	17	120	135	674

948533	3173	3188	TCAACGACCTGGCCCC	20	121	136	675

948534	3174	3189	CTCAACGACCTGGCCC	30	122	137	676

948535	3175	3190	CCTCAACGACCTGGCC	101	123	138	677

948536	3176	3191	CCCTCAACGACCTGGC	57	124	139	678

948537	3278	3293	CTGCAGTTTGTTGACC	45	226	241	679

948538	3299	3314	GCCGATGGAGCTGAAG	7	247	262	680

948539	3300	3315	GGCCGATGGAGCTGAA	104	248	263	681

948540	N/A	N/A	AAGGAAGTCCCGGCCC	81	346	361	682

948541	N/A	N/A	GAAGGAAGTCCCGGCC	101	347	362	683

948542	44750	44765	GATTCCTGAACCGCGG	106	364	379	684

948543	44752	44767	ACGATTCCTGAACCGC	38	366	381	685

948544	44754	44769	TGACGATTCCTGAACC	18	368	383	686

948545	44755	44770	GTGACGATTCCTGAAC	40	369	384	687

948546	44756	44771	GGTGACGATTCCTGAA	42	370	385	688

948547	44758	44773	CGGGTGACGATTCCTG	111	372	387	689

948548	44761	44776	CGCCGGGTGACGATTC	44	375	390	690

948549	44769	44784	TGAGAGGCCGCCGGGT	11	383	398	691

948550	44771	44786	AATGAGAGGCCGCCGG	76	385	400	692

948551	44772	44787	GAATGAGAGGCCGCCG	67	386	401	693

948552	44775	44790	GCAGAATGAGAGGCCG	99	389	404	694

948553	57480	57495	CAAAAACTCGGCATGT	20	427	442	695

948554	57481	57496	GCAAAAACTCGGCATG	15	428	443	696

948555	57490	57505	ACTTGCAGTGCAAAAA	15	437	452	697

948556	57492	57507	GGACTTGCAGTGCAAA	7	439	454	698

948557	57494	57509	TTGGACTTGCAGTGCA	3	441	456	699

948558	57497	57512	TTTTTGGACTTGCAGT	10	444	459	700

948559	57498	57513	CTTTTTGGACTTGCAG	6	445	460	701

948560	57500	57515	AACTTTTTGGACTTGC	3	447	462	702

948561	57501	57516	AAACTTTTTGGACTTG	92	448	463	703

948562	57502	57517	TAAACTTTTTGGACTT	8	449	464	704

948563	57503	57518	GTAAACTTTTTGGACT	26	450	465	705

948564	57504	57519	TGTAAACTTTTTGGAC	15	451	466	706

948565	57524	57539	TGCCGGACTTCATCAA	31	471	486	707

948566	57525	57540	CTGCCGGACTTCATCA	24	472	487	708

948567	57527	57542	TCCTGCCGGACTTCAT	26	474	489	709

948568	57530	57545	ATCTCCTGCCGGACTT	14	477	492	710

948569	57531	57546	AATCTCCTGCCGGACT	18	478	493	711

948570	57535	57550	CTTCAATCTCCTGCCG	29	482	497	712

948571	57597	57612	TCGAAGGTTGATGGGC	89	544	559	713

948572	57598	57613	CTCGAAGGTTGATGGG	88	545	560	714

948573	57599	57614	ACTCGAAGGTTGATGG	75	546	561	715

948574	60706	60721	ATGAGGGTCAAGTTCA	52	579	594	716

948575	60708	60723	CGATGAGGGTCAAGTT	15	581	596	717

948576	60709	60724	TCGATGAGGGTCAAGT	28	582	597	718

948577	60710	60725	GTCGATGAGGGTCAAG	22	583	598	719

948578	60712	60727	AGGTCGATGAGGGTCA	15	585	600	720

948579	60714	60729	GGAGGTCGATGAGGGT	53	587	602	721

948580	60717	60732	CCGGGAGGTCGATGAG	93	590	605	722

948581	60721	60736	ATACCCGGGAGGTCGA	32	594	609	723

948582	60723	60738	TGATACCCGGGAGGTC	34	596	611	724

948583	60725	60740	GGTGATACCCGGGAGG	32	598	613	725

948584	60726	60741	TGGTGATACCCGGGAG	79	599	614	726

948585	60727	60742	TTGGTGATACCCGGGA	27	600	615	727

948586	60729	60744	CCTTGGTGATACCCGG	44	602	617	728

948587	60774	60789	CCTTGATCTGGTACTC	6	647	662	729

948588	60775	60790	TCCTTGATCTGGTACT	20	648	663	730

948589	60777	60792	TGTCCTTGATCTGGTA	12	650	665	731

948590	60779	60794	CATGTCCTTGATCTGG	13	652	667	732

948591	60781	60796	ATCATGTCCTTGATCT	34	654	669	733

948592	60782	60797	GATCATGTCCTTGATC	57	655	670	734

948593	60783	60798	GGATCATGTCCTTGAT	38	656	671	735

948594	60815	60830	AATGAGGCTGCTCTCC	42	688	703	736

948595	60820	60835	GCCAGAATGAGGCTGC	92	693	708	737

948596	60848	60863	GGCCAGGTCCATGTTG	92	721	736	738

948597	N/A	N/A	GGTCCGTAGGCCTTGG	45	775	790	739

948598	62120	62135	CACCGATGGTCCGTAG	49	782	797	740

948599	62192	62207	ACGGGAGCAACTTGTT	76	854	869	741

948600	62196	62211	CTCAACGGGAGCAACT	48	858	873	742

948601	62197	62212	TCTCAACGGGAGCAAC	45	859	874	743

948602	68004	68019	CACGGATGTCCTTCTT	8	923	938	744

948603	68006	68021	TGCACGGATGTCCTTC	25	925	940	745

948604	68007	68022	CTGCACGGATGTCCTT	5	926	941	746

948605	68009	68024	TGCTGCACGGATGTCC	16	928	943	747

948606	68010	68025	GTGCTGCACGGATGTC	20	929	944	748

948607	68011	68026	AGTGCTGCACGGATGT	22	930	945	749

948608	68012	68027	CAGTGCTGCACGGATG	8	931	946	750

948609	68014	68029	GCCAGTGCTGCACGGA	37	933	948	751

948610	68015	68030	TGCCAGTGCTGCACGG	50	934	949	752

948611	68016	68031	CTGCCAGTGCTGCACG	27	935	950	753

948612	68018	68033	AGCTGCCAGTGCTGCA	107	937	952	754

948613	68056	68071	TGCCGGTAGGCCGGGT	122	975	990	755

948614	71653	71668	CCGAAAGTTCTTGTAC	31	1129	1144	756

948615	71656	71671	GGGCCGAAAGTTCTTG	88	1132	1147	757

948616	71657	71672	CGGGCCGAAAGTTCTT	75	1133	1148	758

948617	71658	71673	TCGGGCCGAAAGTTCT	42	1134	1149	759

948618	71660	71675	CGTCGGGCCGAAAGTT	20	1136	1151	760

948619	71683	71698	GGCTTTGGTTTTGCGG	80	1159	1174	761

948620	78729	78744	ATCCACCCCAAACTGC	45	1192	1207	762

948621	78771	78786	AGTGTCCACCTGATCT	27	1234	1249	763

948622	78773	78788	AGAGTGTCCACCTGAT	11	1236	1251	764

948623	78774	78789	CAGAGTGTCCACCTGA	88	1237	1252	765

948624	78775	78790	CCAGAGTGTCCACCTG	74	1238	1253	766

948625	78777	78792	CTCCAGAGTGTCCACC	51	1240	1255	767

948626	78785	78800	CCGGAGAGCTCCAGAG	98	1248	1263	768

948627	78787	78802	CCCCGGAGAGCTCCAG	44	1250	1265	769

948628	78805	78820	TGCGATTGATTCGGGC	11	1268	1283	770

948629	78806	78821	ATGCGATTGATTCGGG	3	1269	1284	771

948630	78810	78825	GAAGATGCGATTGATT	10	1273	1288	772

948631	78811	78826	GGAAGATGCGATTGAT	3	1274	1289	773

948632	78812	78827	TGGAAGATGCGATTGA	9	1275	1290	774

948633	78813	78828	GTGGAAGATGCGATTG	8	1276	1291	775

948634	78829	78844	CAAATGGGAACCGCTC	60	1292	1307	776

948335	78830	78845	TCAAATGGGAACCGCT	65	1293	1308	777

948636	78831	78846	CTCAAATGGGAACCGC	81	1294	1309	778

948637	78832	78847	GCTCAAATGGGAACCG	58	1295	1310	779

948638	78833	78848	AGCTCAAATGGGAACC	43	1296	1311	780

948639	78837	78852	CACCAGCTCAAATGGG	70	1300	1315	781

948640	80420	80435	CTCCATGGATGTTCTT	42	1367	1382	782

948641	81059	81074	TGAAAAGCCCGGTCCT	38	1385	1400	783

948642	81060	81075	GTGAAAAGCCCGGTCC	37	1386	1401	784

948643	81062	81077	GGGTGAAAAGCCCGGT	120	1388	1403	785

948644	81077	81092	CGAATGCCAAGTCCGG	84	1403	1418	786

948645	81078	81093	TCGAATGCCAAGTCCG	28	1404	1419	787

948646	81080	81095	CCTCGAATGCCAAGTC	13	1406	1421	788

948647	81081	81096	GCCTCGAATGCCAAGT	73	1407	1422	789

948648	81084	81099	ATGGCCTCGAATGCCA	94	1410	1425	790

948649	81088	81103	CACAATGGCCTCGAAT	35	1414	1429	791

948650	81104	81119	TGACGACCTGCTTTTT	15	1430	1445	792

948651	81105	81120	TTGACGACCTGCTTTT	21	1431	1446	793

948652	81108	81123	AGCTTGACGACCTGCT	84	1434	1449	794

948653	81110	81125	TCAGCTTGACGACCTG	61	1436	1451	795

948654	81112	81127	TTTCAGCTTGACGACC	11	1438	1453	796

948655	81114	81129	TCTTTCAGCTTGACGA	27	1440	1455	797

948656	81116	81131	GCTCTTTCAGCTTGAC	3	1442	1457	798

948657	81117	81132	GGCTCTTTCAGCTTGA	59	1443	1458	799

948658	81118	81133	GGGCTCTTTCAGCTTG	73	1444	1459	800

948659	81120	81135	CAGGGCTCTTTCAGCT	75	1446	1461	801

948660	81132	81147	ACACATTTCAGACAGG	3	1458	1473	802

948661	81134	81149	CGACACATTTCAGACA	7	1460	1475	803

948662	81135	81150	TCGACACATTTCAGAC	10	1461	1476	804

948663	81136	81151	GTCGACACATTTCAGA	51	1462	1477	805

948664	81138	81153	AGGTCGACACATTTCA	3	1464	1479	806

948665	81140	81155	CCAGGTCGACACATTT	6	1466	1481	807

948666	81141	81156	ACCAGGTCGACACATT	8	1467	1482	808

948667	81142	81157	AACCAGGTCGACACAT	34	1468	1483	809

948668	81146	81161	GGATAACCAGGTCGAC	34	1472	1487	810

948669	81148	81163	CTGGATAACCAGGTCG	43	1474	1489	811

948670	81149	81164	CCTGGATAACCAGGTC	111	1475	1490	812

948671	81150	81165	TCCTGGATAACCAGGT	100	1476	1491	813

948672	81152	81167	GCTCCTGGATAACCAG	51	1478	1493	814

948673	81159	81174	TTGATTAGCTCCTGGA	2	1485	1500	815

948674	81161	81176	TATTGATTAGCTCCTG	8	1487	1502	816

948675	81162	81177	GTATTGATTAGCTCCT	2	1488	1503	817

948676	81163	81178	TGTATTGATTAGCTCC	1	1489	1504	818

948677	81164	81179	CTGTATTGATTAGCTC	2	1490	1505	819

948678	81165	81180	ACTGTATTGATTAGCT	7	1491	1506	820

948679	81168	81183	CTAACTGTATTGATTA	56	1494	1509	821

948680	81169	81184	CCTAACTGTATTGATT	44	1495	1510	822

948681	81171	81186	TGCCTAACTGTATTGA	25	1497	1512	823

948682	81177	81192	GTACACTGCCTAACTG	23	1503	1518	824

948683	81180	81195	CTGGTACACTGCCTAA	14	1506	1521	825

948684	81181	81196	ACTGGTACACTGCCTA	8	1507	1522	826

948685	81183	81198	TTACTGGTACACTGCC	2	1509	1524	827

948686	81184	81199	CTTACTGGTACACTGC	1	1510	1525	828

948687	N/A	N/A	GCTTACTGGTACACTG	6	1511	1526	829

948688	N/A	N/A	AGCTTACTGGTACACT	26	1512	1527	830

948689	N/A	N/A	GAGCTTACTGGTACAC	12	1513	1528	831

948690	N/A	N/A	ACTGAGCTTACTGGTA	47	1516	1531	832

948691	N/A	N/A	AACTGAGCTTACTGGT	24	1517	1532	833

948692	N/A	N/A	GGAACTGAGCTTACTG	10	1519	1534	834

948693	N/A	N/A	AGGAACTGAGCTTACT	12	1520	1535	835

948694	N/A	N/A	TAGGAACTGAGCTTAC	17	1521	1536	836

948695	N/A	N/A	GTAGGAACTGAGCTTA	15	1522	1537	837

948696	83484	83499	GGGTAGGAACTGAGCT	28	1524	1539	838

948697	83500	83515	CTCCTCTCGCAACCGG	56	1540	1555	839

948698	83503	83518	TGTCTCCTCTCGCAAC	44	1543	1558	840

948699	83504	83519	CTGTCTCCTCTCGCAA	27	1544	1559	841

948700	83506	83521	CTCTGTCTCCTCTCGC	5	1546	1561	842

948701	83507	83522	GCTCTGTCTCCTCTCG	8	1547	1562	843

948702	83508	83523	CGCTCTGTCTCCTCTC	5	1548	1563	844

948703	83510	83525	TTCGCTCTGTCTCCTC	5	1550	1565	845

948704	83514	83529	ACGATTCGCTCTGTCT	69	1554	1569	846

948705	83515	83530	GACGATTCGCTCTGTC	91	1555	1570	847

948706	83516	83531	TGACGATTCGCTCTGT	37	1556	1571	848

948707	83521	83536	AGTGGTGACGATTCGC	5	1561	1576	849

948708	83523	83538	TAAGTGGTGACGATTC	25	1563	1578	850

948709	83525	83540	TGTAAGTGGTGACGAT	21	1565	1580	851

948710	83526	83541	ATGTAAGTGGTGACGA	27	1566	1581	852

948711	83527	83542	GATGTAAGTGGTGACG	37	1567	1582	853

948712	83530	83545	CCGGATGTAAGTGGTG	60	1570	1585	854

948713	83543	83558	TCCCCTCCCGTTCCCG	40	1583	1598	855

948714	83545	83560	TCTCCCCTCCCGTTCC	30	1585	1600	856

948715	83546	83561	TTCTCCCCTCCCGTTC	43	1586	1601	857

948716	83547	83562	GTTCTCCCCTCCCGTT	14	1587	1602	858

948717	N/A	N/A	GCAGAAGAATCTGGTC	27	1607	1622	859

948718	87287	87302	GATCAGCAGAAGAATC	126	1612	1627	860

948719	87289	87304	TCGATCAGCAGAAGAA	70	1614	1629	861

948720	87291	87306	TGTCGATCAGCAGAAG	27	1616	1631	862

948721	87292	87307	ATGTCGATCAGCAGAA	51	1617	1632	863

948722	87293	87308	AATGTCGATCAGCAGA	41	1618	1633	864

948723	87295	87310	TCAATGTCGATCAGCA	17	1620	1635	865

948724	87297	87312	GCTCAATGTCGATCAG	14	1622	1637	866

948725	87298	87313	TGCTCAATGTCGATCA	41	1623	1638	867

948726	87301	87316	GACTGCTCAATGTCGA	91	1626	1641	868

948727	87303	87318	AGGACTGCTCAATGTC	28	1628	1643	869

948728	87305	87320	GTAGGACTGCTCAATG	136	1630	1645	870

948729	87306	87321	TGTAGGACTGCTCAAT	2	1631	1646	871

948730	87307	87322	ATGTAGGACTGCTCAA	18	1632	1647	872

948731	87309	87324	TGATGTAGGACTGCTC	6	1634	1649	873

948732	87310	87325	TTGATGTAGGACTGCT	13	1635	1650	874

948733	87312	87327	TGTTGATGTAGGACTG	39	1637	1652	875

948734	87313	87328	GTGTTGATGTAGGACT	22	1638	1653	876

948735	87314	87329	CGTGTTGATGTAGGAC	3	1639	1654	877

948736	N/A	N/A	GGCATTGGCAAACCCG	106	1672	1687	878

948737	97293	97308	CATCAGGCTGATGTTG	43	1765	1780	879

948738	97296	97311	TTTCATCAGGCTGATG	90	1768	1783	880

948739	97340	97355	AGTGACTCGGCAGTCA	100	1812	1827	881

948740	97342	97357	ACAGTGACTCGGCAGT	25	1814	1829	882

948741	97343	97358	GACAGTGACTCGGCAG	13	1815	1830	883

948742	97344	97359	GGACAGTGACTCGGCA	33	1816	1831	884

948743	97346	97361	CAGGACAGTGACTCGG	23	1818	1833	885

948744	97347	97362	CCAGGACAGTGACTCG	36	1819	1834	886

948745	97348	97363	ACCAGGACAGTGACTC	23	1820	1835	887

948746	97349	97364	TACCAGGACAGTGACT	93	1821	1836	888

948747	105014	105029	TCACGGATCTTGAGGT	27	1884	1899	889

948748	105016	105031	CATCACGGATCTTGAG	52	1886	1901	890

948749	105017	105032	ACATCACGGATCTTGA	43	1887	1902	891

948750	105018	105033	CACATCACGGATCTTG	19	1888	1903	892

948751	105021	105036	CTCCACATCACGGATC	60	1891	1906	893

948752	105024	105039	CTTCTCCACATCACGG	4	1894	1909	894

948753	105061	105076	TGAAGATGGCGAAGAC	52	1931	1946	895

948754	105062	105077	TTGAAGATGGCGAAGA	101	1932	1947	896

948755	105063	105078	GTTGAAGATGGCGAAG	83	1933	1948	897

948756	108794	108809	CCGCAGGTCCTTGTAG	60	1966	1981	898

948757	108796	108811	TGCCGCAGGTCCTTGT	42	1968	1983	899

948758	108809	108824	GGCCAGCTCGATCTGC	85	1981	1996	900

948759	108855	108870	GGAGGAACGAGGCCTT	80	2027	2042	901

948760	108856	108871	CGGAGGAACGAGGCCT	107	2028	2043	902

948761	108858	108873	CTCGGAGGAACGAGGC	111	2030	2045	903

948762	108864	108879	CGCCAGCTCGGAGGAA	125	2036	2051	904

948763	110079	110094	GAGAAGGTGTTCTCCT	98	2094	2109	905

948764	110082	110097	ATGGAGAAGGTGTTCT	112	2097	2112	906

948765	110158	110173	GGACTTGTTGATGATG	9	2173	2188	907

948766	110159	110174	TGGACTTGTTGATGAT	10	2174	2189	908

948767	110161	110176	GATGGACTTGTTGATG	12	2176	2191	909

948768	110162	110177	GGATGGACTTGTTGAT	8	2177	2192	910

948769	110163	110178	CGGATGGACTTGTTGA	10	2178	2193	911

948770	110164	110179	GCGGATGGACTTGTTG	36	2179	2194	912

948771	110165	110180	CGCGGATGGACTTGTT	78	2180	2195	913

948772	110173	110188	CATGAGGTCGCGGATG	25	2188	2203	914

948773	110174	110189	GCATGAGGTCGCGGAT	4	2189	2204	915

948774	110176	110191	TGGCATGAGGTCGCGG	9	2191	2206	916

948775	114037	114052	TGGATGAAGGCCTTCG	6	2238	2253	917

948776	114065	114080	GGAGTATAGGTAGGCC	15	2266	2281	918

948777	114067	114082	GAGGAGTATAGGTAGG	7	2268	2283	919

948778	114070	114085	GCCGAGGAGTATAGGT	51	2271	2286	920

948779	114071	114086	TGCCGAGGAGTATAGG	17	2272	2287	921

948780	114152	114167	CTTGAGGGCATGGTAC	48	2353	2368	922

948781	114225	114240	TGTCATCGACAGGCGG	32	2426	2441	923

948782	115139	115154	GGACACCGGTCGGCGC	103	2482	2497	924

948783	115140	115155	TGGACACCGGTCGGCG	43	2483	2498	925

948784	115145	115160	TATGCTGGACACCGGT	33	2488	2503	926

948785	115147	115162	TGTATGCTGGACACCG	22	2490	2505	927

948786	115148	115163	GTGTATGCTGGACACC	78	2491	2506	928

948787	115149	115164	GGTGTATGCTGGACAC	79	2492	2507	929

948788	115212	115227	CGGGAACAGGAATCAG	13	2555	2570	930

948789	115213	115228	ACGGGAACAGGAATCA	19	2556	2571	931

948790	115217	115232	CCCCACGGGAACAGGA	101	2560	2575	932

948791	115235	115250	CGAGAAGGAGGCTGCT	45	2578	2593	933

948792	115236	115251	CCGAGAAGGAGGCTGC	37	2579	2594	934

948793	115237	115252	GCCGAGAAGGAGGCTG	124	2580	2595	935

948794	115279	115294	GCAAACACGCTCTGGG	6	2622	2637	936

948795	115280	115295	GGCAAACACGCTCTGG	11	2623	2638	937

948796	115281	115296	TGGCAAACACGCTCTG	31	2624	2639	938

948797	115282	115297	TTGGCAAACACGCTCT	23	2625	2640	939

948798	115284	115299	TGTTGGCAAACACGCT	15	2627	2642	940

948799	115286	115301	ACTGTTGGCAAACACG	66	2629	2644	941

948800	115287	115302	CACTGTTGGCAAACAC	39	2630	2645	942

948801	115288	115303	TCACTGTTGGCAAACA	28	2631	2646	943

948802	115289	115304	GTCACTGTTGGCAAAC	17	2632	2647	944

948803	115290	115305	GGTCACTGTTGGCAAA	16	2633	2648	945

948804	115291	115306	AGGTCACTGTTGGCAA	9	2634	2649	946

948805	115293	115308	AGAGGTCACTGTTGGC	3	2636	2651	947

948806	115294	115309	AAGAGGTCACTGTTGG	7	2637	2652	948

948807	115295	115310	GAAGAGGTCACTGTTG	19	2638	2653	949

948808	115297	115312	GGGAAGAGGTCACTGT	13	2640	2655	950

948809	115321	115336	CGAGATGGGATCTGAG	33	2664	2679	951

948810	115322	115337	CCGAGATGGGATCTGA	18	2665	2680	952

948811	115323	115338	GCCGAGATGGGATCTG	28	2666	2681	953

948812	115327	115342	ACTGGCCGAGATGGGA	13	2670	2685	954

948813	115328	115343	AACTGGCCGAGATGGG	20	2671	2686	955

948814	115330	115345	CGAACTGGCCGAGATG	27	2673	2688	956

948815	115331	115346	CCGAACTGGCCGAGAT	46	2674	2689	957

948816	115333	115348	ATCCGAACTGGCCGAG	35	2676	2691	958

948817	115334	115349	GATCCGAACTGGCCGA	105	2677	2692	959

948818	115991	116006	GCCGGCTGGGCGCAGC	124	2735	2750	960

948819	116000	116015	TAATGGTGGGCCGGCT	83	2744	2759	961

948820	116002	116017	GATAATGGTGGGCCGG	96	2746	2761	962

948821	116003	116018	GGATAATGGTGGGCCG	73	2747	2762	963

948822	116004	116019	CGGATAATGGTGGGCC	41	2748	2763	964

948823	116014	116029	CTCGGCTGGGCGGATA	109	2758	2773	965

948824	116023	116038	CAGGGATGGCTCGGCT	48	2767	2782	966

948825	116028	116043	TCGAGCAGGGATGGCT	84	2772	2787	967

948826	116029	116044	GTCGAGCAGGGATGGC	135	2773	2788	968

948827	116034	116049	GCCTAGTCGAGCAGGG	84	2778	2793	969

948828	116055	116070	CGAGAGCACGCCCCCC	37	2799	2814	970

948829	116058	116073	CCCCGAGAGCACGCCC	73	2802	2817	971

948830	116092	116107	ACTGAAGCTCCTGCGC	75	2836	2851	972

948831	116109	116124	GCGGAGGGCCCCAGAC	94	2853	2868	973

948832	116163	116178	GTTAAGGAAGAGGCCA	27	2907	2922	974

948833	116210	116225	TGCGGTGTCCAGCCAG	42	2954	2969	975

948834	116212	116227	AGTGCGGTGTCCAGCC	24	2956	2971	976

948835	116213	116228	CAGTGCGGTGTCCAGC	9	2957	2972	977

948836	116214	116229	GCAGTGCGGTGTCCAG	11	2958	2973	978

948837	116216	116231	GCGCAGTGCGGTGTCC	60	2960	2975	979

948838	116217	116232	TGCGCAGTGCGGTGTC	58	2961	2976	980

948839	116221	116236	CCTTTGCGCAGTGCGG	57	2965	2980	981

948840	116222	116237	CCCTTTGCGCAGTGCG	21	2966	2981	982

948841	116224	116239	GCCCCTTTGCGCAGTG	104	2968	2983	983

948842	116225	116240	GGCCCCTTTGCGCAGT	98	2969	2984	984

948843	116261	116276	GTGCAACACCCCAGCG	58	3005	3020	985

948844	116266	116281	CCAAAGTGCAACACCC	31	3010	3025	986

948845	116267	116282	CCCAAAGTGCAACACC	82	3011	3026	987

948846	116288	116303	TGCCACCCTGAGACTC	62	3032	3047	988

948847	116310	116325	TCAAGGGTTCTGGTCC	10	3054	3069	989

948848	116311	116326	GTCAAGGGTTCTGGTC	28	3055	3070	990

948849	116313	116328	GTGTCAAGGGTTCTGG	8	3057	3072	991

948850	116315	116330	TGGTGTCAAGGGTTCT	6	3059	3074	992

948851	116316	116331	ATGGTGTCAAGGGTTC	5	3060	3075	993

948852	116317	116332	GATGGTGTCAAGGGTT	6	3061	3076	994

948853	116319	116334	AGGATGGTGTCAAGGG	13	3063	3078	995

948854	116320	116335	CAGGATGGTGTCAAGG	11	3064	3079	996

948855	116321	116336	TCAGGATGGTGTCAAG	13	3065	3080	997

948856	116323	116338	ATTCAGGATGGTGTCA	13	3067	3082	998

948857	116325	116340	TCATTCAGGATGGTGT	20	3069	3084	999

948858	116327	116342	CCTCATTCAGGATGGT	82	3071	3086	1000

948859	116351	116366	GTAGAGTCCCCCCCAG	22	3095	3110	1001

948860	116353	116368	TGGTAGAGTCCCCCCC	44	3097	3112	1002

948861	116354	116369	TTGGTAGAGTCCCCCC	19	3098	3113	1003

948862	116355	116370	CTTGGTAGAGTCCCCC	9	3099	3114	1004

948863	116357	116372	ACCTTGGTAGAGTCCC	4	3101	3116	1005

948864	116358	116373	GACCTTGGTAGAGTCC	94	3102	3117	1006

948865	116361	116376	GAAGACCTTGGTAGAG	6	3105	3120	1007

948866	116363	116378	AAGAAGACCTTGGTAG	17	3107	3122	1008

948867	116364	116379	CAAGAAGACCTTGGTA	60	3108	3123	1009

948868	116365	116380	CCAAGAAGACCTTGGT	62	3109	3124	1010

948869	116366	116381	CCCAAGAAGACCTTGG	70	3110	3125	1011

948870	116367	116382	GCCCAAGAAGACCTTG	60	3111	3126	1012

948871	116384	116399	CTACATGGGCTTTCCC	85	3128	3143	1013

948872	116386	116401	CCCTACATGGGCTTTC	98	3130	3145	1014

948873	116399	116414	TATAGAAGGCCTGCCC	100	3143	3158	1015

948874	116401	116416	CTTATAGAAGGCCTGC	20	3145	3160	1016

948875	116402	116417	ACTTATAGAAGGCCTG	25	3146	3161	1017

948876	116403	116418	CACTTATAGAAGGCCT	34	3147	3162	1018

948877	116405	116420	CGCACTTATAGAAGGC	57	3149	3164	1019

948878	116406	116421	CCGCACTTATAGAAGG	83	3150	3165	1020

948879	116450	116465	CCCTGCACCCCAGCAA	128	3194	3209	1021

948880	116454	116469	ATACCCCTGCACCCCA	18	3198	3213	1022

948881	116456	116471	ATATACCCCTGCACCC	45	3200	3215	1023

948882	116457	116472	GATATACCCCTGCACC	11	3201	3216	1024

948883	116458	116473	TGATATACCCCTGCAC	13	3202	3217	1025

948884	116459	116474	TTGATATACCCCTGCA	9	3203	3218	1026

948885	116460	116475	GTTGATATACCCCTGC	4	3204	3219	1027

948886	116465	116480	GGGAAGTTGATATACC	14	3209	3224	1028

948887	116466	116481	TGGGAAGTTGATATAC	20	3210	3225	1029

948888	116468	116483	AATGGGAAGTTGATAT	59	3212	3227	1030

948889	116469	116484	TAATGGGAAGTTGATA	49	3213	3228	1031

948890	116470	116485	CTAATGGGAAGTTGAT	15	3214	3229	1032

948891	116472	116487	TGCTAATGGGAAGTTG	12	3216	3231	1033

948892	116473	116488	CTGCTAATGGGAAGTT	4	3217	3232	1034

948893	116475	116490	TCCTGCTAATGGGAAG	2	3219	3234	1035

948894	116476	116491	CTCCTGCTAATGGGAA	86	3220	3235	1036

948895	116479	116494	GAGCTCCTGCTAATGG	94	3223	3238	1037

948896	116498	116513	GGCCAGGCTTGCCGCT	99	3242	3257	1038

948897	116512	116527	TACCGAGCCCACTGGG	103	3256	3271	1039

948898	116515	116530	CACTACCGAGCCCACT	82	3259	3274	1040

948899	116517	116532	GGCACTACCGAGCCCA	64	3261	3276	1041

948900	116520	116535	CTGGGCACTACCGAGC	92	3264	3279	1042

948901	116525	116540	GCCAGCTGGGCACTAC	149	3269	3284	1043

948902	116526	116541	TGCCAGCTGGGCACTA	130	3270	3285	1044

948903	116539	116554	TACACCTCAGGCCTGC	28	3283	3298	1045

948904	116540	116555	GTACACCTCAGGCCTG	98	3284	3299	1046

948905	116542	116557	ATGTACACCTCAGGCC	77	3286	3301	1047

948906	116543	116558	TATGTACACCTCAGGC	9	3287	3302	1048

948907	116545	116560	ACTATGTACACCTCAG	5	3289	3304	1049

948908	116546	116561	GACTATGTACACCTCA	16	3290	3305	1050

948909	116548	116563	AGGACTATGTACACCT	125	3292	3307	1051

948910	116549	116564	AAGGACTATGTACACC	5	3293	3308	1052

948911	116551	116566	GGAAGGACTATGTACA	11	3295	3310	1053

948912	116552	116567	GGGAAGGACTATGTAC	28	3296	3311	1054

948913	116553	116568	CGGGAAGGACTATGTA	16	3297	3312	1055

948914	116554	116569	CCGGGAAGGACTATGT	60	3298	3313	1056

948915	116555	116570	GCCGGGAAGGACTATG	40	3299	3314	1057

948916	116560	116575	ATATGGCCGGGAAGGA	21	3304	3319	1058

948917	116562	116577	TAATATGGCCGGGAAG	2	3306	3321	1059

948918	116563	116578	TTAATATGGCCGGGAA	9	3307	3322	1060

948919	116564	116579	GTTAATATGGCCGGGA	16	3308	3323	1061

948920	116566	116581	TGGTTAATATGGCCGG	37	3310	3325	1062

948921	116567	116582	GTGGTTAATATGGCCG	45	3311	3326	1063

948922	116568	116583	TGTGGTTAATATGGCC	10	3312	3327	1064

948923	116570	116585	TGTGTGGTTAATATGG	3	3314	3329	1065

948924	116571	116586	CTGTGTGGTTAATATG	4	3315	3330	1066

948925	116572	116587	GCTGTGTGGTTAATAT	7	3316	3331	1067

948926	116574	116589	AGGCTGTGTGGTTAAT	3	3318	3333	1068

948927	116618	116633	GGCCTAGCAAAGGCAC	118	3362	3377	1069

948928	116631	116646	GCCAACGGCTCCGGGC	119	3375	3390	1070

948929	116637	116652	GCCCGGGCCAACGGCT	96	3381	3396	1071

948930	116645	116660	CAAGGCCGGCCCGGGC	131	3389	3404	1072

948931	116647	116662	GGCAAGGCCGGCCCGG	104	3391	3406	1073

948932	116652	116667	AATAGGGCAAGGCCGG	65	3396	3411	1074

948933	116653	116668	GAATAGGGCAAGGCCG	126	3397	3412	1075

948934	116654	116669	GGAATAGGGCAAGGCC	22	3398	3413	1076

948935	116656	116671	GAGGAATAGGGCAAGG	7	3400	3415	1077

948936	116657	116672	AGAGGAATAGGGCAAG	4	3401	3416	1078

948937	116658	116673	GAGAGGAATAGGGCAA	6	3402	3417	1079

948938	116660	116675	AGGAGAGGAATAGGGC	8	3404	3419	1080

948939	116705	116720	ACATAGCCCAAGCCCA	54	3449	3464	1081

948940	116706	116721	CACATAGCCCAAGCCC	72	3450	3465	1082

948941	116707	116722	CCACATAGCCCAAGCC	90	3451	3466	1083

948942	116709	116724	ACCCACATAGCCCAAG	71	3453	3468	1084

948943	116767	116782	ACCCATCAGGGAGGCA	93	3511	3526	1085

948944	116790	116805	CAGAGAGAGGCCGCCC	43	3534	3549	1086

948945	116793	116808	CCTCAGAGAGAGGCCG	134	3537	3552	1087

948946	116814	116829	AGCGAGGAGTGGGTGA	43	3558	3573	1088

948947	116817	116832	CTGAGCGAGGAGTGGG	57	3561	3576	1089

948948	116818	116833	ACTGAGCGAGGAGTGG	34	3562	3577	1090

948949	116820	116835	AAACTGAGCGAGGAGT	64	3564	3579	1091

948950	116821	116836	CAAACTGAGCGAGGAG	7	3565	3580	1092

948951	116823	116838	GTCAAACTGAGCGAGG	5	3567	3582	1093

948952	116824	116839	GGTCAAACTGAGCGAG	5	3568	3583	1094

948953	116825	116840	TGGTCAAACTGAGCGA	9	3569	3584	1095

948954	116827	116842	AGTGGTCAAACTGAGC	5	3571	3586	1096

948955	116828	116843	CAGTGGTCAAACTGAG	64	3572	3587	1097

948956	116829	116844	ACAGTGGTCAAACTGA	73	3573	3588	1098

948957	116830	116845	TACAGTGGTCAAACTG	69	3574	3589	1099

948958	116831	116846	TTACAGTGGTCAAACT	34	3575	3590	1100

948959	116833	116848	ACTTACAGTGGTCAAA	17	3577	3592	1101

948960	116834	116849	CACTTACAGTGGTCAA	11	3578	3593	1102

948961	116835	116850	GCACTTACAGTGGTCA	4	3579	3594	1103

948962	116836	116851	GGCACTTACAGTGGTC	45	3580	3595	1104

948963	116837	116852	AGGCACTTACAGTGGT	5	3581	3596	1105

948964	116841	116856	GTGCAGGCACTTACAG	43	3585	3600	1106

948965	116844	116859	AGAGTGCAGGCACTTA	7	3588	3603	1107

948966	116846	116861	ACAGAGTGCAGGCACT	8	3590	3605	1108

948967	116847	116862	TACAGAGTGCAGGCAC	12	3591	3606	1109

948968	116848	116863	ATACAGAGTGCAGGCA	6	3592	3607	1110

948969	116850	116865	GAATACAGAGTGCAGG	6	3594	3609	1111

948970	116851	116866	AGAATACAGAGTGCAG	2	3595	3610	1112

948971	116852	116867	TAGAATACAGAGTGCA	1	3596	3611	1113

948972	116854	116869	AATAGAATACAGAGTG	6	3598	3613	1114

948973	3394	3409	GCTCACCGGCCCACGA	72	N/A	N/A	1115

948974	3500	3515	CGCGGGCACGCAAGCG	144	N/A	N/A	1116

948975	3607	3622	CGTGACACCGCCCCTG	71	N/A	N/A	1117

948976	3715	3730	ATCTACCCCAGACCCA	63	N/A	N/A	1118

948977	3725	3740	CAGAAGAGGGATCTAC	99	N/A	N/A	1119

948978	3776	3791	CAATAGATATCTAGAT	144	N/A	N/A	1120

948979	3778	3793	CCCAATAGATATCTAG	59	N/A	N/A	1121

948980	3815	3830	AGATACAAGAACAAGC	67	N/A	N/A	1122

948981	3934	3949	AGCTAGCCAGACAGCA	146	N/A	N/A	1123

948982	4010	4025	CAACAGAGGGACTGCT	85	N/A	N/A	1124

948983	4035	4050	ACAGTAGAGGCCAGCT	96	N/A	N/A	1125

948984	4130	4145	CGGAAGAGAGGAGCCA	84	N/A	N/A	1126

948985	4135	4150	TCACACGGAAGAGAGG	63	N/A	N/A	1127

948986	4142	4157	GAAGAATTCACACGGA	50	N/A	N/A	1128

948987	4262	4277	GTCCAAACCTCCTCCT	63	N/A	N/A	1129

948988	4315	4330	TCCTGAAGTAAGGTAC	96	N/A	N/A	1130

948989	4326	4341	TCGGAAAGGAATCCTG	40	N/A	N/A	1131

948990	4371	4386	AATTTGTAGGCTGTAG	21	N/A	N/A	1132

948991	4388	4403	TCCAAAGGAGGGCCCC	140	N/A	N/A	1133

948992	4476	4491	CACCGCGGCTTCTCAA	38	N/A	N/A	1134

948993	4624	4639	GAGAAGTGGCCGCCCA	75	N/A	N/A	1135

948994	4649	4664	CCACAAATGAGTAGTT	49	N/A	N/A	1136

948995	4730	4745	CATTATCCCTTGACAA	115	N/A	N/A	1137

948996	4742	4757	ATCAAGAAGGGACATT	85	N/A	N/A	1138

948997	4838	4853	GGTGATCACAGTCCCT	72	N/A	N/A	1139

948998	4938	4953	AGAGACAAGGTAGCAA	45	N/A	N/A	1140

948999	4950	4965	AGGCAATGAGCTAGAG	39	N/A	N/A	1141

949000	5049	5064	GAAGATGGTCACTCAA	47	N/A	N/A	1142

949001	5051	5066	AGGAAGATGGTCACTC	32	N/A	N/A	1143

949002	5162	5177	GTCAACCAAGACTTAG	48	N/A	N/A	1144

949003	5545	5560	TAGAATGGCATCTTGT	28	N/A	N/A	1145

949004	5710	5725	CCGGATCTGGTTCTGT	88	N/A	N/A	1146

949005	5810	5825	AACCAGCCAGATAAAC	94	N/A	N/A	1147

949006	5872	5887	GCTTAAGTGTGTAACG	8	N/A	N/A	1148

949007	5873	5888	GGCTTAAGTGTGTAAC	10	N/A	N/A	1149

949008	5898	5913	GATTATAGCAATGCCT	22	N/A	N/A	1150

949009	5940	5955	GCTCAGCTATCTGAGC	107	N/A	N/A	1151

949010	6074	6089	CCAGACACCATGGTAG	33	N/A	N/A	1152

949011	6150	6165	ATATAATGAGCAACTC	47	N/A	N/A	1153

949012	6174	6189	GTGAGACTCGGTGTAA	10	N/A	N/A	1154

949013	6298	6313	GTGGTAACGCGCGCCT	41	N/A	N/A	1155

949014	6455	6470	AATACTCTTAACGAGG	95	N/A	N/A	1156

949015	6556	6571	ACCTTATCTCTTACAC	63	N/A	N/A	1157

949016	6697	6712	TCCAGGCTCGGCGCAA	83	N/A	N/A	1158

949017	6752	6767	AGCTATAGGGCTGAAC	129	N/A	N/A	1159

949018	6816	6831	CGCAGGTAAAACAAAA	38	N/A	N/A	1160

949019	6817	6832	ACGCAGGTAAAACAAA	56	N/A	N/A	1161

949020	6821	6836	AGCTACGCAGGTAAAA	66	N/A	N/A	1162

949021	6824	6839	AAAAGCTACGCAGGTA	27	N/A	N/A	1163

949022	6870	6885	CCGGATATTGACTGGA	46	N/A	N/A	1164

949023	6891	6906	TGTCTTAGAGCCACAA	131	N/A	N/A	1165

949024	6906	6921	CTCTATAGATCTGAGT	123	N/A	N/A	1166

949025	6918	6933	GACAACTCAGAACTCT	93	N/A	N/A	1167

949026	7031	7046	TACCACCCTCAGCTTA	69	N/A	N/A	1168

949027	7131	7146	CGCCAGGTGAAACTGA	86	N/A	N/A	1169

949028	7155	7170	CCACAAGGGACAAAGG	89	N/A	N/A	1170

949029	7170	7185	TCGAAGCCATTACTGC	89	N/A	N/A	1171

949030	7248	7263	TGCTAGGGAGTAGTGG	47	N/A	N/A	1172

949031	7273	7288	GCAAAATGATCTAGGC	26	N/A	N/A	1173

949032	7328	7343	CCGCAGACAAGCCAGG	99	N/A	N/A	1174

949033	7351	7366	CCAAAGTTCTATCTGC	15	N/A	N/A	1175

949034	7395	7410	TGACAAGGGTCAAGAG	91	N/A	N/A	1176

949035	7400	7415	CTAAATGACAAGGGTC	22	N/A	N/A	1177

949036	7402	7417	GGCTAAATGACAAGGG	47	N/A	N/A	1178

949037	7645	7660	GTATAAGACCCTATCT	83	N/A	N/A	1179

949038	7849	7864	ATTTGAGGCGAGGAGG	58	N/A	N/A	1180

949050	8567	8582	TTTTAAAGAAGACGGG	51	N/A	N/A	1181

949051	8584	8599	TGCTATTATAACTCCT	15	N/A	N/A	1182

949052	8671	8686	GAACAACACCTAATGT	47	N/A	N/A	1183

949053	8675	8690	GAACGAACAACACCTA	8	N/A	N/A	1184

949054	8678	8693	AATGAACGAACAACAC	22	N/A	N/A	1185

949055	8705	8720	AGTAAAAGTACTCAGG	8	N/A	N/A	1186

949056	8740	8755	TGGTAACTTGCTCAGG	16	N/A	N/A	1187

949057	8773	8788	GCTGAAAGGCCAGCTC	83	N/A	N/A	1188

949058	8949	8964	CTACAGGTGTCACCAT	100	N/A	N/A	1189

949059	9093	9108	ACTTATCGCCTTTTTT	11	N/A	N/A	1190

949061	9613	9628	CTCCACTGGCCTGCTA	4	N/A	N/A	1191

949062	9724	9739	AGGGACAGCAGTAGCG	69	N/A	N/A	1192

949063	9830	9845	AGAAAAATGGCACGGG	41	N/A	N/A	1193

949064	9883	9898	TAAAAACGCAGCTGGA	79	N/A	N/A	1194

949065	9971	9986	TGCACTGGTGCAGCAC	115	N/A	N/A	1195

949066	10072	10087	GGCTCTAAACTAATAA	143	N/A	N/A	1196

949067	10260	10275	TCGGGATTCACTATGT	28	N/A	N/A	1197

949068	10379	10394	CGCTTACTGCAAGTTC	40	N/A	N/A	1198

949069	10481	10496	CCTAACCGAGCCTTGC	78	N/A	N/A	1199

949070	10675	10690	ACAACAATTTGCTACG	45	N/A	N/A	1200

949071	10677	10692	TCACAACAATTTGCTA	23	N/A	N/A	1201

949072	11266	11281	TGACACCAAACTGAGC	92	N/A	N/A	1202

949073	11356	11371	AAAGAAACTAAGGCCG	87	N/A	N/A	1203

949074	11384	11399	TAATGATGAGTTCCTT	14	N/A	N/A	1204

949075	11392	11407	CAACAGAGTAATGATG	54	N/A	N/A	1205

949076	11496	11511	CGATATTTCAGGATCT	88	N/A	N/A	1206

949077	11597	11612	CTCATCACCGTACACA	9	N/A	N/A	1207

949078	11881	11896	CCCTATCATTGGTTTT	55	N/A	N/A	1208

949079	11984	11999	TAACAGGTCTTGTGCA	55	N/A	N/A	1209

949080	12093	12108	ACGATTCACATGTGGG	11	N/A	N/A	1210

949081	12141	12156	AACCTAATCCCCAAAC	104	N/A	N/A	1211

949082	12195	12210	AGCACTTGGACAGCCT	36	N/A	N/A	1212

949083	12200	12215	GAAAAAGCACTTGGAC	31	N/A	N/A	1213

949084	12214	12229	CAATAAGTAACTCTGA	52	N/A	N/A	1214

949085	12318	12333	ATCTACAACTGTCCCC	25	N/A	N/A	1215

949086	12555	12570	AATAATGAGACCCCAC	145	N/A	N/A	1216

949087	12667	12682	TAAATCTAACCTAGGC	126	N/A	N/A	1217

949088	12778	12793	CAGAAGTTTGGCAGAT	5	N/A	N/A	1218

949089	12788	12803	TCAAATCGGGCAGAAG	22	N/A	N/A	1219

949090	12858	12873	AAATTACGGTTTACAT	71	N/A	N/A	1220

949091	12859	12874	TAAATTACGGTTTACA	20	N/A	N/A	1221

949092	12878	12893	TTGTAGTTGGCAATCC	12	N/A	N/A	1222

949093	12892	12907	GGCTTAATTACAGCTT	86	N/A	N/A	1223

949094	12901	12916	CACAATAAGGGCTTAA	10	N/A	N/A	1224

949095	12902	12917	ACACAATAAGGGCTTA	10	N/A	N/A	1225

949096	12947	12962	CCCTATAAATATCTCA	23	N/A	N/A	1226

949097	13037	13052	ACGCACCCGGAAGCAG	71	N/A	N/A	1227

949098	13140	13155	AAACAGTCATCCAGCT	105	N/A	N/A	1228

949099	13208	13223	TGGCACACGGTGATTC	57	N/A	N/A	1229

949100	13215	13230	GCACATGTGGCACACG	27	N/A	N/A	1230

949101	13273	13288	AGGGATGCTAAGAGCA	69	N/A	N/A	1231

949102	13281	13296	TATAAAGCAGGGATGC	117	N/A	N/A	1232

949103	13391	13406	CCATATGTGTGGCTCT	21	N/A	N/A	1233

949104	13520	13535	GCATATCCAAAGCCAC	38	N/A	N/A	1234

949105	13635	13650	CATTTAACTGTTCACC	28	N/A	N/A	1235

949106	13812	13827	CGACAGACAAGTACCA	46	N/A	N/A	1236

949107	13952	13967	GCATACAGCCTTTTTT	38	N/A	N/A	1237

949108	13953	13968	CGCATACAGCCTTTTT	15	N/A	N/A	1238

949109	13955	13970	ACCGCATACAGCCTTT	9	N/A	N/A	1239

949110	13956	13971	CACCGCATACAGCCTT	46	N/A	N/A	1240

949111	13959	13974	AACCACCGCATACAGC	50	N/A	N/A	1241

949112	13961	13976	TGAACCACCGCATACA	40	N/A	N/A	1242

949113	14262	14277	TGCAACCTTCAAGTAA	94	N/A	N/A	1243

949114	14390	14405	GCAAACCGCTTCTCAC	74	N/A	N/A	1244

949115	14492	14507	CCCCAAGTCCTGGCGG	99	N/A	N/A	1245

949116	14629	14644	GAAAAGTCGGCATTCC	74	N/A	N/A	1246

949117	14757	14772	TGACAGTGGTGCTCCC	66	N/A	N/A	1247

949118	15000	15015	TGGCATGTGGTTGTTG	26	N/A	N/A	1248

949119	15165	15180	AACAAATATATGCACG	20	N/A	N/A	1249

949120	15370	15385	TCGCACCTCGGGATTC	92	N/A	N/A	1250

949121	15507	15522	GTGAAATTCACTCCAT	40	N/A	N/A	1251

949122	15612	15627	GGAAGATGTCCTTGCT	49	N/A	N/A	1252

949123	15643	15658	ATAGATGGAGCAAATC	77	N/A	N/A	1253

949124	15647	15662	TTATATAGATGGAGCA	32	N/A	N/A	1254

949125	15708	15723	ACGCAGATAGACACCC	53	N/A	N/A	1255

949126	15712	15727	GCCAACGCAGATAGAC	93	N/A	N/A	1256

949127	15777	15792	TGCCATACAGGGCCTC	116	N/A	N/A	1257

949128	15859	15874	GAAAGAGCAGTGGTGC	73	N/A	N/A	1258

949129	15973	15988	GGGCATGGAGGTGTCA	90	N/A	N/A	1259

949130	16025	16040	CGCGAAAGTGTGTCAC	64	N/A	N/A	1260

949131	16117	16132	TATCACCCAGCAGGCA	75	N/A	N/A	1261

949132	16275	16290	GGCCAGCTGGTAGGCC	114	N/A	N/A	1262

949134	16714	16729	GTCAAGCTGAGCCAGT	52	N/A	N/A	1263

949135	16815	16830	ACAGAAGCCGGGCTTC	127	N/A	N/A	1264

949136	16926	16941	ATCAATTTTCACCCAG	19	N/A	N/A	1265

949137	16930	16945	GTGCATCAATTTTCAC	127	N/A	N/A	1266

949138	17244	17259	AAAGAGTTTAAGGGCG	57	N/A	N/A	1267

949139	17282	17297	CCACAACGAGTAAACA	15	N/A	N/A	1268

949140	17615	17630	GGACAGGGTGAATTGT	62	N/A	N/A	1269

949141	17723	17738	AGTCACACCTAGAAAT	32	N/A	N/A	1270

949142	17758	17773	GACTTATAGGCTCACT	12	N/A	N/A	1271

949143	17763	17778	TACTAGACTTATAGGC	26	N/A	N/A	1272

949144	17765	17780	GTTACTAGACTTATAG	19	N/A	N/A	1273

949145	17766	17781	TGTTACTAGACTTATA	40	N/A	N/A	1274

949146	17834	17849	ATATAGCAGGGACCTG	70	N/A	N/A	1275

949147	17838	17853	CTAAATATAGCAGGGA	36	N/A	N/A	1276

949148	17840	17855	CACTAAATATAGCAGG	24	N/A	N/A	1277

949149	18278	18293	TTTGAGCCCGAGAGGG	46	N/A	N/A	1278

949150	18486	18501	TTATTTCTAGGCAGGG	13	N/A	N/A	1279

949151	18524	18539	GTAAATTTGAAGCTTA	144	N/A	N/A	1280

949152	18583	18598	AATTAGAGGACACTGG	19	N/A	N/A	1281

949153	18665	18680	GCTTTAACTCCTAGAC	61	N/A	N/A	1282

949154	19040	19055	GTATACCAAGGTTAAT	56	N/A	N/A	1283

949155	19050	19065	CTTAATAGTAGTATAC	96	N/A	N/A	1284

949156	19458	19473	CACACGAGGTCTCACT	44	N/A	N/A	1285

949157	19461	19476	CTACACACGAGGTCTC	72	N/A	N/A	1286

949158	19663	19678	TACATTCTGGCTTTGG	12	N/A	N/A	1287

949159	19763	19778	TCTCAGTATGTATACT	12	N/A	N/A	1288

949160	19814	19829	TGTCCAGTCCTAAGAC	101	N/A	N/A	1289

949161	19816	19831	AATGTCCAGTCCTAAG	81	N/A	N/A	1290

949162	19817	19832	CAATGTCCAGTCCTAA	19	N/A	N/A	1291

949163	19818	19833	GCAATGTCCAGTCCTA	4	N/A	N/A	1292

949164	19820	19835	ATGCAATGTCCAGTCC	13	N/A	N/A	1293

949165	19821	19836	GATGCAATGTCCAGTC	70	N/A	N/A	1294

949166	19822	19837	AGATGCAATGTCCAGT	13	N/A	N/A	1295

949167	19824	19839	CTAGATGCAATGTCCA	12	N/A	N/A	1296

949168	19843	19858	GCTTAAGGAGATCCGA	17	N/A	N/A	1297

949169	19844	19859	GGCTTAAGGAGATCCG	85	N/A	N/A	1298

949170	19872	19887	GGCTGAGTTGCTTCCC	79	N/A	N/A	1299

949171	19935	19950	TTCCAGATAAGCCCTA	32	N/A	N/A	1300

949172	19986	20001	CACCAATGGCCTTCTT	42	N/A	N/A	1301

949173	20328	20343	GAAGATGGCTGGACGA	34	N/A	N/A	1302

949174	20330	20345	CAGAAGATGGCTGGAC	34	N/A	N/A	1303

949175	20378	20393	CAATAATTTATCAGTC	68	N/A	N/A	1304

949176	20440	20455	GGGAAACCTAGCAAGC	80	N/A	N/A	1305

949177	20487	20502	CCCAAAGGATAGAGTA	23	N/A	N/A	1306

949178	20735	20750	TTACATGAGTCTAGGA	50	N/A	N/A	1307

949179	20737	20752	TATTACATGAGTCTAG	68	N/A	N/A	1308

949180	20740	20755	GATTATTACATGAGTC	49	N/A	N/A	1309

949181	20748	20763	GGCAAGAGGATTATTA	53	N/A	N/A	1310

949182	20844	20859	AGCAACTTTAGGGAGA	5	N/A	N/A	1311

949183	20899	20914	GAACAGACAGTTCAGG	76	N/A	N/A	1312

949184	20947	20962	AAGATATGTGGTTTTG	35	N/A	N/A	1313

949185	21341	21356	GAGCATCACCAGCCCA	127	N/A	N/A	1314

949186	21403	21418	TAGAAATGAGTGGGCG	116	N/A	N/A	1315

949187	21416	21431	AGCTACAGCAATATAG	43	N/A	N/A	1316

949188	21449	21464	ACTCACATGGCCTGAT	81	N/A	N/A	1317

949189	21491	21506	CTAGACTAGACAAAAC	91	N/A	N/A	1318

949190	21514	21529	ATATATATGTGGAGGC	4	N/A	N/A	1319

949191	21561	21576	CTAATACACAATGATC	34	N/A	N/A	1320

949192	21591	21606	CAAAAAACTAATACCG	105	N/A	N/A	1321

949193	21698	21713	AACCAGTACCAGCTGG	98	N/A	N/A	1322

949194	21758	21773	AGAAACTGTACTGGTC	25	N/A	N/A	1323

949195	21845	21860	GGATATTCTTAACAGG	26	N/A	N/A	1324

949196	21854	21869	AGCTAGGAAGGATATT	83	N/A	N/A	1325

949197	22009	22024	AAGTATGCGCTGCCAT	38	N/A	N/A	1326

949198	22018	22033	TGGGACTGTAAGTATG	63	N/A	N/A	1327

949199	22133	22148	GGATATAGGGTCTCCC	138	N/A	N/A	1328

949200	22243	22258	CATGAGGTAGGACAGC	54	N/A	N/A	1329

949201	22251	22266	GAAAAAGCCATGAGGT	100	N/A	N/A	1330

949202	22301	22316	CCACAGGGTTATCTGG	119	N/A	N/A	1331

949203	22345	22360	AAGGAGGCTTTTAAGC	92	N/A	N/A	1332

949204	22446	22461	AGGAATGACAAGTGGT	17	N/A	N/A	1333

949205	22466	22481	CGCATAGGTGGAACAA	58	N/A	N/A	1334

949206	22478	22493	CAAGAAACCAAACGCA	81	N/A	N/A	1335

949207	22638	22653	AATATACGGGCATGTT	82	N/A	N/A	1336

949208	22639	22654	AAATATACGGGCATGT	110	N/A	N/A	1337

949209	22641	22656	AAAAATATACGGGCAT	104	N/A	N/A	1338

949210	22643	22658	ACAAAAATATACGGGC	67	N/A	N/A	1339

949211	22644	22659	TACAAAAATATACGGG	33	N/A	N/A	1340

949212	22780	22795	TCAAACAAATAGGCCG	86	N/A	N/A	1341

949213	22862	22877	ATGCAAGGGACTGCGC	66	N/A	N/A	1342

949214	22883	22898	ACCTATCGCATGCCAG	17	N/A	N/A	1343

949215	23000	23015	GTTCAGGTCAGAGCCT	51	N/A	N/A	1344

949216	23297	23312	TCTCAATGAAAGTCTC	10	N/A	N/A	1345

949217	23548	23563	GTAACAACCGCTGGGA	27	N/A	N/A	1346

949218	23555	23570	TTAAAGAGTAACAACC	75	N/A	N/A	1347

949219	23715	23730	GCCCAGCTAACTAACT	108	N/A	N/A	1348

949220	24040	24055	GACTACAGTACAGGCG	74	N/A	N/A	1349

949221	24201	24216	AGAACAAGGGAACACG	80	N/A	N/A	1350

949222	24210	24225	CCCCATAGAAGAACAA	138	N/A	N/A	1351

949223	24289	24304	GATTAGGTATTATGGT	12	N/A	N/A	1352

949224	24296	24311	TCAAAAGGATTAGGTA	25	N/A	N/A	1353

949225	24315	24330	ATCCACTGAATAGTAT	32	N/A	N/A	1354

949226	24364	24379	CTACAAAATCAGATAG	91	N/A	N/A	1355

949227	24438	24453	GACATTAGTGGCTTCC	35	N/A	N/A	1356

949228	24561	24576	AACCAGCATGGATGAA	87	N/A	N/A	1357

949229	24782	24797	TGCAACCTTGCCCCCC	11	N/A	N/A	1358

949230	24934	24949	TAGTATTCAAAAAACG	80	N/A	N/A	1359

949231	25044	25059	CAAATTAAATACACCC	62	N/A	N/A	1360

949232	25058	25073	GAATACTATTCTAACA	118	N/A	N/A	1361

949233	25210	25225	CACCACAATTTGCTCT	63	N/A	N/A	1362

949234	25305	25320	TCAGAAAAGGAGGGTA	109	N/A	N/A	1363

949235	25321	25336	TTCCACTTCAAGAACC	87	N/A	N/A	1364

949236	25696	25711	TGTAAGCAACTGTATT	49	N/A	N/A	1365

949237	25788	25803	TGAGAAGATATGCTGA	26	N/A	N/A	1366

949238	25822	25837	AACCTATGTATCAAAG	87	N/A	N/A	1367

949239	25925	25940	CAAAGTAGTTCCTCCA	34	N/A	N/A	1368

949240	25927	25942	TACAAAGTAGTTCCTC	68	N/A	N/A	1369

949241	26042	26057	CCCAAGAGTAAAATAC	105	N/A	N/A	1370

949242	26084	26099	ACGAAACCAATAAGAC	104	N/A	N/A	1371

949243	26090	26105	GAAAATACGAAACCAA	99	N/A	N/A	1372

949244	26133	26148	TAATAATGGATGGAAC	84	N/A	N/A	1373

949245	26155	26170	TGGTATCAGGCAAGAA	35	N/A	N/A	1374

949246	27033	27048	CATGATGCTGCAATCA	97	N/A	N/A	1375

949247	27177	27192	CACAAGCCTGCGGACC	110	N/A	N/A	1376

949248	27191	27206	GAGCATAGCAGACACA	65	N/A	N/A	1377

949249	27254	27269	GGCAACATCACTCTAA	62	N/A	N/A	1378

949250	27289	27304	CAAAGAGCCTTGGCAC	111	N/A	N/A	1379

949251	27390	27405	CAGGACTAGTTCACTA	78	N/A	N/A	1380

949252	27473	27488	ATGAAATTGGGATAAC	60	N/A	N/A	1381

949253	27509	27524	ATGAAGGCTGCCACTT	103	N/A	N/A	1382

949254	27575	27590	AGCAAGATCATTGGGT	65	N/A	N/A	1383

949255	27634	27649	GGAAACCTTGGACTCC	91	N/A	N/A	1384

949256	27761	27776	GTCTATTGGCACACAG	38	N/A	N/A	1385

949257	27775	27790	GATTAAGTCATCCGGT	70	N/A	N/A	1386

949258	27872	27887	GGGCACTGCACAAAGC	86	N/A	N/A	1387

949259	27975	27990	AGTCAGCGGAGACTGA	98	N/A	N/A	1388

949260	28212	28227	CGGCAAAAGAAGCTGG	133	N/A	N/A	1389

949261	28320	28335	AGCTAGAGCATCAGCT	122	N/A	N/A	1390

949262	28971	28986	CTGGAAACAGTCTCGC	29	N/A	N/A	1391

949263	29116	29131	AGCCGAAGTGTAGTGG	51	N/A	N/A	1392

949264	29258	29273	GGCCAAGTCTCCCATC	117	N/A	N/A	1393

949265	29370	29385	CCGGACCCGTTCTTGA	85	N/A	N/A	1394

949266	29501	29516	AGGTAGTGACCTCCAG	114	N/A	N/A	1395

949267	29557	29572	ACCCACAGTCTGCATT	97	N/A	N/A	1396

949268	29591	29606	GCCCAAGTGAACTAGA	94	N/A	N/A	1397

949269	29625	29640	ATGTATGGATTTCCAG	62	N/A	N/A	1398

949270	29642	29657	TTACAGGGTGGTCAGC	82	N/A	N/A	1399

949271	29721	29736	GTAAAACGGTTGTGGT	30	N/A	N/A	1400

949272	29739	29754	CAACAGGTGTCCTGCT	64	N/A	N/A	1401

949273	29842	29857	TGTCATCCTAACTGCT	57	N/A	N/A	1402

949274	29943	29958	AGAAACTGGCCGGCCC	132	N/A	N/A	1403

949275	30052	30067	TTTATAGTCTCCAACG	34	N/A	N/A	1404

949276	30178	30193	ACGGACTGGATTATGC	27	N/A	N/A	1405

949277	30248	30263	GGCTAAATCAATCTTG	60	N/A	N/A	1406

949278	30282	30297	CGATAATGACCTAAAT	84	N/A	N/A	1407

949279	30283	30298	CCGATAATGACCTAAA	8	N/A	N/A	1408

949280	30406	30421	GAATATCAACACCTTG	35	N/A	N/A	1409

949281	30508	30523	AGTGATATGCTGCCTA	39	N/A	N/A	1410

949282	30705	30720	GGTTTATTTGCACCTC	39	N/A	N/A	1411

949283	30830	30845	TACCAGGGACTCAACT	111	N/A	N/A	1412

949284	30933	30948	GGCCAGACATTGCTGT	142	N/A	N/A	1413

949285	31273	31288	CACATTGGTGGTCAAG	98	N/A	N/A	1414

949286	31373	31388	CTCATAACATTTGACG	45	N/A	N/A	1415

949287	31708	31723	AAAAAAGCCGGCACGG	120	N/A	N/A	1416

949288	32027	32042	AAAATTGGTTGAGTGC	40	N/A	N/A	1417

949289	32161	32176	GTAAAACTCAGGCTGA	81	N/A	N/A	1418

949290	32192	32207	CAAAAGAGAACATTCG	64	N/A	N/A	1419

949291	32264	32279	TGTTATGTGGGAATGA	35	N/A	N/A	1420

949292	32420	32435	AGGGACACAACAGGCT	85	N/A	N/A	1421

949293	32560	32575	CACAATCCACTCCTGG	100	N/A	N/A	1422

949294	32561	32576	CCACAATCCACTCCTG	83	N/A	N/A	1423

949295	32615	32630	TTGCAGAACAATGGGC	105	N/A	N/A	1424

949296	32697	32712	GCGCAGGGTCCCGTAT	95	N/A	N/A	1425

949297	32719	32734	TATTAGAGTGGCAGCC	86	N/A	N/A	1426

949298	32799	32814	TGCCACTCACAACTGG	104	N/A	N/A	1427

949299	32852	32867	CGCCATAGGAAGCTGC	111	N/A	N/A	1428

949300	32895	32910	GAGCAGGCGGGCAGCT	82	N/A	N/A	1429

949301	32899	32914	AACAGAGCAGGCGGGC	92	N/A	N/A	1430

949302	32900	32915	TAACAGAGCAGGCGGG	103	N/A	N/A	1431

949303	32903	32918	CATTAACAGAGCAGGC	119	N/A	N/A	1432

949304	32904	32919	GCATTAACAGAGCAGG	46	N/A	N/A	1433

949305	33006	33021	AGCCACATATGCCTCC	91	N/A	N/A	1434

949306	33148	33163	TGTTATGGGCCCTCCT	82	N/A	N/A	1435

949307	33277	33292	TCCAAGGCTGTCAGCG	106	N/A	N/A	1436

949308	33391	33406	CACGAGGCTGCAGACC	40	N/A	N/A	1437

949309	33454	33469	CCGCAGGTAGGTGCCA	56	N/A	N/A	1438

949310	33488	33503	GCCCAAGCCTTACAGT	117	N/A	N/A	1439

949311	33512	33527	AGCCATTGGTCACCCA	43	N/A	N/A	1440

949312	33618	33633	CCGCACCCACCTCAGG	132	N/A	N/A	1441

949313	33644	33659	CACCAATGAGGCAGAC	69	N/A	N/A	1442

949314	33725	33740	GTGGACTGAGGTTCAG	101	N/A	N/A	1443

949315	33826	33841	CTCTAACCCCATCTCG	81	N/A	N/A	1444

949316	33927	33942	TCGGCAGGAGCCCACA	108	N/A	N/A	1445

949317	34028	34043	CCATATGCTCAGCACA	30	N/A	N/A	1446

949318	34029	34044	ACCATATGCTCAGCAC	45	N/A	N/A	1447

949319	34159	34174	GATAAGCCCACAGAGG	107	N/A	N/A	1448

949320	34164	34179	CAACAGATAAGCCCAC	83	N/A	N/A	1449

949321	34281	34296	ACAGACCACAAGGCTC	65	N/A	N/A	1450

949322	34283	34298	GAACAGACCACAAGGC	82	N/A	N/A	1451

949323	34407	34422	CAGGAAGGCGGTAGGG	92	N/A	N/A	1452

949324	34539	34554	TGGGAGGACACTTCCA	121	N/A	N/A	1453

949325	34546	34561	CATGAAATGGGAGGAC	9	N/A	N/A	1454

949326	34561	34576	ACTGACTTGGCCATCC	68	N/A	N/A	1455

949327	34654	34669	CCAGACTGTGGAGCCA	35	N/A	N/A	1456

949328	34763	34778	GCCAAGACTTCTGCAC	129	N/A	N/A	1457

949329	34873	34888	GCCTACCTGGCCATGG	119	N/A	N/A	1458

949330	34995	35010	GCTGACCGGGTTCCCC	148	N/A	N/A	1459

949331	35150	35165	TCCTATCCATGGGCCC	93	N/A	N/A	1460

949332	35178	35193	TACTATTGCATCATTT	33	N/A	N/A	1461

949333	35385	35400	CCCTAGTGAACTTCCC	38	N/A	N/A	1462

949334	35541	35556	GCAAACCTCAGTCTCG	39	N/A	N/A	1463

949335	35921	35936	GCACAATTAAGTAGTT	36	N/A	N/A	1464

949336	35950	35965	GCATAGAGTAGGGCTG	55	N/A	N/A	1465

949337	35955	35970	CCAAAGCATAGAGTAG	57	N/A	N/A	1466

949338	35956	35971	CCCAAAGCATAGAGTA	62	N/A	N/A	1467

949339	36021	36036	CCCATCTGAGCTGTGT	79	N/A	N/A	1468

949340	36170	36185	TCGAATGACATTTCCT	65	N/A	N/A	1469

949341	36211	36226	AGTAAGACAGGGTAAC	136	N/A	N/A	1470

949342	36311	36326	GTGCACCCAGGCTGCC	105	N/A	N/A	1471

949343	36420	36435	TTCCAGCCTCCGTCAT	90	N/A	N/A	1472

949344	36531	36546	CCTAAGGGTTTTCTGG	109	N/A	N/A	1473

949345	36532	36547	CCCTAAGGGTTTTCTG	144	N/A	N/A	1474

949346	36570	36585	CCAGAAGTCTCCTAGG	108	N/A	N/A	1475

949347	36647	36662	CTAAAAGTTCCCAGAC	86	N/A	N/A	1476

949348	36757	36772	GGAGACGCAGCTCTCA	103	N/A	N/A	1477

949349	36892	36907	GGACAAGCACCCCTCT	26	N/A	N/A	1478

949350	36996	37011	CCCGACCCCGGCCAAT	96	N/A	N/A	1479

949351	37100	37115	GTTCACTACAGACATC	71	N/A	N/A	1480

949352	37105	37120	TTCTAGTTCACTACAG	92	N/A	N/A	1481

949353	37243	37258	CATAGTACCCCACACA	56	N/A	N/A	1482

949354	37300	37315	TTGTAGTGTGGAATAT	40	N/A	N/A	1483

949355	37364	37379	CGCCCAAGAGAACACA	104	N/A	N/A	1484

949356	37481	37496	CTCCACTTTGTGTCTG	58	N/A	N/A	1485

949357	37583	37598	GCCTTCTGAGCAAACA	90	N/A	N/A	1486

949358	37726	37741	AAAATGGGTCTGCTGG	18	N/A	N/A	1487

949359	37730	37745	CATTAAAATGGGTCTG	56	N/A	N/A	1488

949360	37843	37858	ACCTTAAGTTTCTTGT	70	N/A	N/A	1489

949361	37896	37911	TGCAATGGAGTCCAAA	65	N/A	N/A	1490

949362	37970	37985	CCTTAAGTTTTTTGGT	132	N/A	N/A	1491

949363	38144	38159	TCATGATTATTAACCT	29	N/A	N/A	1492

949364	38160	38175	AGGCATTGAGAACTTT	77	N/A	N/A	1493

949365	38190	38205	AGCTATTCAACACTGA	65	N/A	N/A	1494

949366	38206	38221	CATTAGAGTAGCTAAT	141	N/A	N/A	1495

949367	38246	38261	TTGAATTCCGCATCAT	86	N/A	N/A	1496

949368	38313	38328	AACAAAACAGGACCCG	108	N/A	N/A	1497

949369	38353	38368	GAAGAAAGTCTGTACT	86	N/A	N/A	1498

949370	38488	38503	TTGTAGTGATCAGTTT	54	N/A	N/A	1499

949371	38588	38603	ATCCATGCTGCTATGC	63	N/A	N/A	1500

949372	38688	38703	TTGCACAACAAAGAGC	90	N/A	N/A	1501

949373	38948	38963	GATATATTGCTGGGCA	72	N/A	N/A	1502

949374	38949	38964	AGATATATTGCTGGGC	44	N/A	N/A	1503

949375	39017	39032	ACCTAAGGAAGCTGAC	72	N/A	N/A	1504

949376	39062	39077	TTAAAGCCCAGCAACA	93	N/A	N/A	1505

949377	39088	39103	ACGAAAAATTCAGATC	104	N/A	N/A	1506

949378	39089	39104	GACGAAAAATTCAGAT	83	N/A	N/A	1507

949379	39124	39139	ACAGAATTGTTACAAC	104	N/A	N/A	1508

949380	39156	39171	GTACAGGGTGGACATT	90	N/A	N/A	1509

949381	39162	39177	ACAAAAGTACAGGGTG	66	N/A	N/A	1510

949382	39163	39178	TACAAAAGTACAGGGT	86	N/A	N/A	1511

949383	39164	39179	ATACAAAAGTACAGGG	54	N/A	N/A	1512

949384	39195	39210	AGTCAAGGAGAACATG	63	N/A	N/A	1513

949385	39529	39544	TTGTAGTGGGTCACGC	53	N/A	N/A	1514

949386	39644	39659	CATGATTGCGCCGTTG	17	N/A	N/A	1515

949387	39745	39760	GTAATTCACTGCGGAT	32	N/A	N/A	1516

949388	39877	39892	AAACTTGCAGCAGTGG	67	N/A	N/A	1517

949389	39981	39996	AAATGGACTAGAAGGG	59	N/A	N/A	1518

949390	40014	40029	AGATAGAAATCAGGGC	47	N/A	N/A	1519

949391	40058	40073	TCACAGGGTGGACTTG	53	N/A	N/A	1520

949392	40089	40104	GTAGACACATGTCCTT	49	N/A	N/A	1521

949393	40195	40210	GACTACAGTCTGTTCG	96	N/A	N/A	1522

949394	40295	40310	AACACAGGAGCAGCAT	87	N/A	N/A	1523

949395	40306	40321	AACGATAAAACAACAC	70	N/A	N/A	1524

949396	40409	40424	ACTGAATCACCAGCAG	99	N/A	N/A	1525

949397	40837	40852	GCCTTTACTTAAGATA	3	N/A	N/A	1526

949398	40961	40976	ACGCAGTGTCTCAGGT	125	N/A	N/A	1527

949399	41061	41076	CACAAGTAGCCTTGGC	107	N/A	N/A	1528

949400	41107	41122	TCACAGTGGACACCAG	85	N/A	N/A	1529

949401	41162	41177	TGACAGACCCCAAAGT	121	N/A	N/A	1530

949402	41234	41249	GAACAAGCTACATCAA	100	N/A	N/A	1531

949403	41266	41281	TGGAGAGGGTTCTCTG	94	N/A	N/A	1532

949404	41272	41287	GATAATTGGAGAGGGT	84	N/A	N/A	1533

949405	41393	41408	GCGAACCTTTCTCACT	57	N/A	N/A	1534

949406	41447	41462	AAACAGGATACGAGAC	74	N/A	N/A	1535

949407	41451	41466	GCACAAACAGGATACG	86	N/A	N/A	1536

949408	41497	41512	TAATCCCTTTCCGTCA	92	N/A	N/A	1537

949409	41604	41619	TGTCTTCGAGCCACAC	99	N/A	N/A	1538

949410	41708	41723	CCCTACAGACACTTGC	101	N/A	N/A	1539

949411	41814	41829	CCGTAGGCAGATTCCA	62	N/A	N/A	1540

949412	41862	41877	GCGAAACCAGAGAGAT	88	N/A	N/A	1541

949413	41863	41878	GGCGAAACCAGAGAGA	86	N/A	N/A	1542

949414	41962	41977	AGCTTTTAAACCCCCC	87	N/A	N/A	1543

949415	41984	41999	AGCCAGATTATTAACA	58	N/A	N/A	1544

949416	42064	42079	GATGAGGAGGCTGGTC	78	N/A	N/A	1545

949417	42169	42184	ACGAAGACCCCCCAGG	163	N/A	N/A	1546

949418	42203	42218	CATGAGAAAAGGGATC	97	N/A	N/A	1547

949419	42249	42264	CAACAAGTCAGCCCAA	100	N/A	N/A	1548

949420	42252	42267	ACACAACAAGTCAGCC	78	N/A	N/A	1549

949421	42254	42269	CTACACAACAAGTCAG	28	N/A	N/A	1550

949422	42318	42333	TTCTAGCCCGTCAGCC	90	N/A	N/A	1551

949423	42434	42449	TTCCAGAAGACCTGTT	143	N/A	N/A	1552

949424	42507	42522	TAGCACTACAATCATT	71	N/A	N/A	1553

949425	42544	42559	AACGAGACAGCATCCT	64	N/A	N/A	1554

949426	42577	42592	GCAATTGGTACTGTAC	101	N/A	N/A	1555

949427	42584	42599	ACACAGGGCAATTGGT	144	N/A	N/A	1556

949428	42727	42742	TCGAACTCAACCTCAG	70	N/A	N/A	1557

949429	42860	42875	TGCCTCGGGTTCAAGC	64	N/A	N/A	1558

949430	42983	42998	TGAAAGGTTCTAGGCA	64	N/A	N/A	1559

949431	42984	42999	GTGAAAGGTTCTAGGC	57	N/A	N/A	1560

949432	43017	43032	TCGAAAGGAAATTCTG	89	N/A	N/A	1561

949433	43028	43043	ACATTAAAGGCTCGAA	43	N/A	N/A	1562

949434	43030	43045	GTACATTAAAGGCTCG	59	N/A	N/A	1563

949435	43050	43065	GTAGAAACAGTGGGCT	61	N/A	N/A	1564

949436	43094	43109	GTACACTCAGAAAGGA	91	N/A	N/A	1565

949437	43141	43156	GTGAATACACCAGGAT	76	N/A	N/A	1566

949438	43194	43209	ACTCAGGCACAACCAC	123	N/A	N/A	1567

949439	43864	43879	CGACAGAGCTGTTTAA	47	N/A	N/A	1568

949440	43884	43899	GCGGAATGTAAATTAC	40	N/A	N/A	1569

949441	43918	43933	CTAAAGACCACTAATT	122	N/A	N/A	1570

949442	43920	43935	GACTAAAGACCACTAA	53	N/A	N/A	1571

949443	44005	44020	GTACAGAGCCATGACT	113	N/A	N/A	1572

949444	44161	44176	TAAAAGCCTGGACAAG	77	N/A	N/A	1573

949445	44199	44214	TACATCTGGACTATGG	23	N/A	N/A	1574

949446	44271	44286	GATACCTGGATGTGGG	80	N/A	N/A	1575

949447	44336	44351	GACAAAATAGGATGTG	88	N/A	N/A	1576

949448	44351	44366	AGGAAAGTTATTGATG	63	N/A	N/A	1577

949449	44356	44371	ATACGAGGAAAGTTAT	112	N/A	N/A	1578

949450	44359	44374	AAAATACGAGGAAAGT	80	N/A	N/A	1579

949451	44362	44377	GGTAAAATACGAGGAA	23	N/A	N/A	1580

949452	44375	44390	CCACATGTATATGGGT	85	N/A	N/A	1581

949453	44426	44441	CCATACCCAGGAGATC	57	N/A	N/A	1582

949454	44509	44524	AACCATTTCCCTCTGG	58	N/A	N/A	1583

949455	44624	44639	GGCGACCTCAGCAAAG	85	N/A	N/A	1584

949456	44732	44747	AAGGAAGTCCCTGTGA	106	N/A	N/A	1585

949457	44851	44866	GGCAACCACACATCCA	79	N/A	N/A	1586

949458	44953	44968	ACTGAGCCGTTTTTCA	42	N/A	N/A	1587

949459	45279	45294	CCGAAGTGAGAGGGTC	88	N/A	N/A	1588

949460	45364	45379	CAAAATCGGGTTATCT	66	N/A	N/A	1589

949461	45365	45380	GCAAAATCGGGTTATC	38	N/A	N/A	1590

949462	45412	45427	AAGGATGGATGTGTCA	18	N/A	N/A	1591

949463	45503	45518	TGACATGCGGGAATGA	40	N/A	N/A	1592

949464	45523	45538	AGTAGAAACTGGGACG	43	N/A	N/A	1593

949465	45570	45585	AGCTATAAAACCAGCC	106	N/A	N/A	1594

949466	45603	45618	TACAAAAGAGTATAGC	70	N/A	N/A	1595

949467	45705	45720	CACCACTGCACGGCCT	97	N/A	N/A	1596

949468	45850	45865	AACATAACCCCCATCT	88	N/A	N/A	1597

949469	45957	45972	AGCAATGGTAGAGGGC	113	N/A	N/A	1598

949470	46279	46294	GCAATGGTGGCCGGGC	96	N/A	N/A	1599

949471	46323	46338	TTCAACAGGCTGGGTC	44	N/A	N/A	1600

949472	46330	46345	TACTATATTCAACAGG	18	N/A	N/A	1601

949473	46344	46359	CCACACATAGGCCATA	35	N/A	N/A	1602

949474	46382	46397	ATACACTGAGATCAGG	50	N/A	N/A	1603

949475	46431	46446	GCTGGGATATACGCCC	98	N/A	N/A	1604

949476	46505	46520	ACGGAGACCCTTACCT	129	N/A	N/A	1605

949477	46528	46543	TTGCAGATTAGCAGGG	35	N/A	N/A	1606

949478	46560	46575	ACAAACTGAGTGTTGA	73	N/A	N/A	1607

949479	46562	46577	ACACAAACTGAGTGTT	78	N/A	N/A	1608

949480	46577	46592	CGCAAGAAGACATTAA	54	N/A	N/A	1609

949481	46666	46681	GGCAGCATGGCTAAGC	75	N/A	N/A	1610

949482	46764	46779	ACGAAAGTGAGGAGTC	66	N/A	N/A	1611

949483	46766	46781	GGACGAAAGTGAGGAG	15	N/A	N/A	1612

949484	46863	46878	TACGAAATAGCAGAGC	17	N/A	N/A	1613

949485	46885	46900	GCCCATTGGCCCATCG	154	N/A	N/A	1614

949486	47094	47109	GATCACCCGCCACTGT	105	N/A	N/A	1615

949487	47352	47367	CACCAGGGACTCATTA	124	N/A	N/A	1616

949488	47466	47481	ACACAGTGGTGGTCCC	59	N/A	N/A	1617

949489	47517	47532	TGCTTTAAGGCTAACA	82	N/A	N/A	1618

949490	47599	47614	GGCCTGCGCTCTGTGC	87	N/A	N/A	1619

949491	47700	47715	TGCCCTAAGGCAAGTG	91	N/A	N/A	1620

949492	47797	47812	CAACAGCCACAGGATC	77	N/A	N/A	1621

949493	47803	47818	TTCCACCAACAGCCAC	25	N/A	N/A	1622

949494	47924	47939	AACTATGCATCCACCT	122	N/A	N/A	1623

949495	47927	47942	TGCAACTATGCATCCA	130	N/A	N/A	1624

949496	48220	48235	GGCTAGGTGGTGTGCG	116	N/A	N/A	1625

949497	48501	48516	GAGATTGGAGCCTGGT	33	N/A	N/A	1626

949498	48506	48521	AATAAGAGATTGGAGC	89	N/A	N/A	1627

949499	48507	48522	CAATAAGAGATTGGAG	69	N/A	N/A	1628

949500	48508	48523	GCAATAAGAGATTGGA	13	N/A	N/A	1629

949501	48622	48637	GTGAATCCAACAGACA	70	N/A	N/A	1630

949502	48748	48763	CAACATGCCTTCCAGT	124	N/A	N/A	1631

949503	48869	48884	TGCCACTCCCTATCCC	79	N/A	N/A	1632

949504	48972	48987	GCTTAATGCCTTGCCT	128	N/A	N/A	1633

949505	49052	49067	CCGCAATGTGTTCATT	42	N/A	N/A	1634

949506	49095	49110	GTATACCCCACAAGGC	73	N/A	N/A	1635

949507	49171	49186	GTAAAGGGTGGCAGCC	84	N/A	N/A	1636

949508	49196	49211	AGTCACTTTAACAAGT	57	N/A	N/A	1637

949509	49218	49233	GCTACCAAGGCACAGA	72	N/A	N/A	1638

949510	49265	49280	CATGGCCACGATGGCA	92	N/A	N/A	1639

949511	49297	49312	GAAGAGACAGCATACC	69	N/A	N/A	1640

949512	49403	49418	AGTTATGGTTCACTGG	29	N/A	N/A	1641

949513	49511	49526	ACTCAGAGTTTACGGG	91	N/A	N/A	1642

949514	49613	49628	TTCCGGTGAGCACACC	49	N/A	N/A	1643

949515	49770	49785	GAATCAGGAGGGAGAT	101	N/A	N/A	1644

949516	49888	49903	GGCAAGTGACCAGGTG	55	N/A	N/A	1645

949517	49896	49911	ACGAAGGCGGCAAGTG	6	N/A	N/A	1646

949518	50269	50284	GCAGAGAGAGTCCATT	38	N/A	N/A	1647

949519	50271	50286	GCGCAGAGAGAGTCCA	116	N/A	N/A	1648

949520	50402	50417	GCCCAGTAGGCAAGAA	117	N/A	N/A	1649

949521	50423	50438	AAGTAACCCCCATCAC	94	N/A	N/A	1650

949522	50491	50506	CTAGATACAAGGCTGC	62	N/A	N/A	1651

949523	50510	50525	CGCCAGCTTGGTCTGC	70	N/A	N/A	1652

949524	50597	50612	ACGCAAATATCTCCCA	41	N/A	N/A	1653

949525	50612	50627	CTAATCCGAGGCAATA	59	N/A	N/A	1654

949526	50654	50669	GCACACAACACTTCCC	41	N/A	N/A	1655

949527	50713	50728	GGCAGGATCCACTCAT	111	N/A	N/A	1656

949528	50818	50833	AGGAGAGGAGATCTGC	87	N/A	N/A	1657

949529	50847	50862	ACACAATGACGAGGCT	27	N/A	N/A	1658

949530	50945	50960	TGTTAGTCTCAGGGAA	54	N/A	N/A	1659

949531	50961	50976	ACAAACACGGCTCAGC	77	N/A	N/A	1660

949532	51078	51093	CGACACCCAGCTCCGG	95	N/A	N/A	1661

949533	51207	51222	TTGAAATGCCCAGGTC	37	N/A	N/A	1662

949534	51318	51333	TTCAAAGGAGCTTGTG	100	N/A	N/A	1663

949535	51433	51448	TGCAAGCTGCCACCTT	79	N/A	N/A	1664

949536	51496	51511	TGCTAAACAGTCAATC	62	N/A	N/A	1665

949537	51544	51559	AACCTAGGAGGACATC	62	N/A	N/A	1666

949538	51660	51675	CCCTACTGAAGCTGGA	71	N/A	N/A	1667

949539	51787	51802	AATCACGCTGCCAAGG	112	N/A	N/A	1668

949540	51890	51905	TCAGAGAGGTTCGCAG	57	N/A	N/A	1669

949541	51993	52008	CGGCAAACTGCAAAGC	67	N/A	N/A	1670

949542	52114	52129	CAACAAGCCACTGAGC	108	N/A	N/A	1671

949543	52204	52219	TAACAATCCACTGGCC	116	N/A	N/A	1672

949544	52243	52258	GCTCAAGTGCTGGAGA	89	N/A	N/A	1673

949545	52494	52509	CAAGATCCTACATGTA	79	N/A	N/A	1674

949546	52597	52612	TAAAAGCTTCCACCAG	119	N/A	N/A	1675

949547	52610	52625	CCCAATAGAGGTTTAA	100	N/A	N/A	1676

949548	52802	52817	TCTCATACTTCGGCCT	111	N/A	N/A	1677

949549	52918	52933	CAACAGAGGTTTTTTG	67	N/A	N/A	1678

949550	52944	52959	GTGAAAGTAAATGCTG	48	N/A	N/A	1679

949551	53030	53045	AACAGAACCACACCAG	216	N/A	N/A	1680

949552	53066	53081	CCAGAAAACCGTTTCT	93	N/A	N/A	1681

949553	53509	53524	ATGCAGGCTGGGAACC	88	N/A	N/A	1682

949554	53548	53563	TGCCAGACTTTGGAGC	92	N/A	N/A	1683

949555	53611	53626	CCCAAGTCCGCCCAGT	90	N/A	N/A	1684

949556	53712	53727	CTGAATCTAGGTGTCT	54	N/A	N/A	1685

949557	53847	53862	CTTGAATGTCCATGCC	60	N/A	N/A	1686

949558	53863	53878	ATATAGAGAGCTTTAC	104	N/A	N/A	1687

949559	53866	53881	CTAATATAGAGAGCTT	51	N/A	N/A	1688

949560	53867	53882	TCTAATATAGAGAGCT	78	N/A	N/A	1689

949561	53909	53924	CCCCATAACACTTACT	98	N/A	N/A	1690

949562	54006	54021	AGCTACTCTCTCAAGT	53	N/A	N/A	1691

949563	54011	54026	GGGCAAGCTACTCTCT	126	N/A	N/A	1692

949564	54127	54142	AGCTACGGTCTGAGCA	114	N/A	N/A	1693

949565	54237	54252	AGTCAGAGGGCCTGCC	58	N/A	N/A	1694

949566	54341	54356	GCTCACTTTGGATTGT	48	N/A	N/A	1695

949567	54452	54467	GTCTACAAAGCCCAGC	67	N/A	N/A	1696

949568	54562	54577	AAAGAGACCATCTCCC	91	N/A	N/A	1697

949569	54673	54688	GGGAAACACTCGGCCG	142	N/A	N/A	1698

949570	54784	54799	CAATGAGGACATCTGC	59	N/A	N/A	1699

949571	54787	54802	GGGCAATGAGGACATC	54	N/A	N/A	1700

949572	54890	54905	AGGAACTTCGCCTCTT	112	N/A	N/A	1701

949573	54924	54939	TCCTAAGCCAGATAGC	49	N/A	N/A	1702

949574	55043	55058	TCTAAGATCCCCATTT	106	N/A	N/A	1703

949575	55144	55159	TTTCACCGTCCCATCA	72	N/A	N/A	1704

949576	55247	55262	AATAACACAGCCATGC	84	N/A	N/A	1705

949577	55348	55363	ACCCACCCCGGAGATG	100	N/A	N/A	1706

949578	55383	55398	TAATAAGTCACACTGG	47	N/A	N/A	1707

949579	55384	55399	GTAATAAGTCACACTG	107	N/A	N/A	1708

949580	55469	55484	CTATGATCTATGATCA	115	N/A	N/A	1709

949581	55678	55693	TCCGTATAAGATGTGA	34	N/A	N/A	1710

949582	56054	56069	CTATTATCCAGCACTG	56	N/A	N/A	1711

949583	56111	56126	AGCTACTGTAGTGATG	81	N/A	N/A	1712

949584	56167	56182	TCGAATTTTCAGAGTA	26	N/A	N/A	1713

949585	56229	56244	ACGGAAGGTGGCTGCC	61	N/A	N/A	1714

949586	56251	56266	GCAAATATAAGGCATG	60	N/A	N/A	1715

949587	56268	56283	CACAGATTGAGGACAA	47	N/A	N/A	1716

949588	56319	56334	TGACAAGCAGTGTGGG	48	N/A	N/A	1717

949589	56371	56386	GGTATCTGAAAGTCAC	9	N/A	N/A	1718

949590	56530	56545	CCACAAGGTAGAGGGA	154	N/A	N/A	1719

949591	56538	56553	CAAGGAGCCCACAAGG	114	N/A	N/A	1720

949592	56633	56648	GCCCAAGTCCTTGTGG	105	N/A	N/A	1721

949593	56691	56706	GATTAAGGTGATGCTG	61	N/A	N/A	1722

949594	56692	56707	TGATTAAGGTGATGCT	41	N/A	N/A	1723

949595	56734	56749	AGGAAGCACCAGGATA	23	N/A	N/A	1724

949596	56860	56875	CATATTATGAATCCCC	66	N/A	N/A	1725

949597	56863	56878	TGACATATTATGAATC	105	N/A	N/A	1726

949598	56979	56994	ATGACAAGGGTCTTGG	17	N/A	N/A	1727

949599	57373	57388	TAATGACCCAGGCTGG	121	N/A	N/A	1728

949600	57619	57634	GCCTACCGTGTGGCGA	104	N/A	N/A	1729

949601	57654	57669	CCAGAAACGGTCAGTG	132	N/A	N/A	1730

949602	57722	57737	TGTGAATGGGCACATG	102	N/A	N/A	1731

949603	57840	57855	CAAACAACTTCTTGGC	82	N/A	N/A	1732

949604	57948	57963	GAAGACTGGTTCTGGC	60	N/A	N/A	1733

949605	57953	57968	TTACAGAAGACTGGTT	91	N/A	N/A	1734

949606	58891	58906	GGCAAGGTGGCTAAAA	123	N/A	N/A	1735

949607	58993	59008	AGGAACCCATCATGGT	95	N/A	N/A	1736

949608	59092	59107	TGACAAGGTAGGTTTT	57	N/A	N/A	1737

949609	59093	59108	TTGACAAGGTAGGTTT	25	N/A	N/A	1738

949610	59102	59117	CAAAACTGGTTGACAA	47	N/A	N/A	1739

949611	59104	59119	TGCAAAACTGGTTGAC	58	N/A	N/A	1740

949612	59272	59287	GTGAAGTGCAGTAGCT	88	N/A	N/A	1741

949613	59302	59317	TAAGATAGAGGGTCTT	35	N/A	N/A	1742

949614	59399	59414	GCTAACAATTTAAGGA	60	N/A	N/A	1743

949615	59741	59756	CTGAGAGGCATACAAA	66	N/A	N/A	1744

949616	59846	59861	GGAGAAGAAGTGAGCC	109	N/A	N/A	1745

949617	60067	60082	ACAAATGGATGAGGCC	129	N/A	N/A	1746

949618	60068	60083	GACAAATGGATGAGGC	54	N/A	N/A	1747

949619	60379	60394	GACAAACAGATGAGGC	74	N/A	N/A	1748

949620	60427	60442	GAATAAATTTCAGCCC	100	N/A	N/A	1749

949621	60483	60498	TTCAATGGCACAGTTC	73	N/A	N/A	1750

949622	60586	60601	TCAGACTTGAGGATGT	32	N/A	N/A	1751

949623	60639	60654	GCACAAACAGGCCCGC	99	N/A	N/A	1752

949624	60700	60715	GTCAAGTTCAACACTG	25	N/A	N/A	1753

949625	60903	60918	GCTCAGGGTTACCTTG	81	N/A	N/A	1754

949626	61033	61048	AGGAAGTGCCATGTTC	64	N/A	N/A	1755

949627	61134	61149	TCCCACTCTGGCAACA	79	N/A	N/A	1756

949628	61204	61219	AGCTAGATGGGACCTA	102	N/A	N/A	1757

949629	61255	61270	GAAGTTCCCGTTCTGA	47	N/A	N/A	1758

949630	61336	61351	AAATAGACGATCTCAC	87	N/A	N/A	1759

949631	61340	61355	AGGGAAATAGACGATC	96	N/A	N/A	1760

949632	61344	61359	TCGAAGGGAAATAGAC	101	N/A	N/A	1761

949633	61382	61397	GTAGAAGGATCTGACT	114	N/A	N/A	1762

949634	61384	61399	CAGTAGAAGGATCTGA	123	N/A	N/A	1763

949635	61480	61495	GTGAAGATCTCTGGTG	88	N/A	N/A	1764

949636	61485	61500	CCAGAGTGAAGATCTC	119	N/A	N/A	1765

949637	61587	61602	CTGCAGATCAACTGCT	104	N/A	N/A	1766

949638	61636	61651	CCACAAGTCATTCCAG	71	N/A	N/A	1767

949639	61706	61721	TTCCAGGACTACCTCT	76	N/A	N/A	1768

949640	61813	61828	GGCTTTGGGAAACCTC	48	N/A	N/A	1769

949641	61839	61854	TATTAAGGACAACCTC	98	N/A	N/A	1770

949642	61840	61855	TTATTAAGGACAACCT	81	N/A	N/A	1771

949643	61842	61857	TGTTATTAAGGACAAC	125	N/A	N/A	1772

949644	61889	61904	TGGCACACGGCAAATA	96	N/A	N/A	1773

949645	61904	61919	TCATAAGGGTGAGCCT	70	N/A	N/A	1774

949646	61915	61930	TCACAGGGTGGTCATA	68	N/A	N/A	1775

949647	61937	61952	GAAGATGGTCACAGCC	51	N/A	N/A	1776

949648	61944	61959	GTCGAATGAAGATGGT	38	N/A	N/A	1777

949649	62105	62120	GGCCTGAAAGAGCCCA	112	N/A	N/A	1778

949650	62252	62267	GATAAGGACCCCACCC	108	N/A	N/A	1779

949651	62328	62343	AAAGAAGGAGTTACGG	88	N/A	N/A	1780

949652	62378	62393	ACGAATGCATACAAAT	72	N/A	N/A	1781

949653	62384	62399	TGAAAAACGAATGCAT	107	N/A	N/A	1782

949654	62491	62506	CACCAGTGGCTGTCTG	63	N/A	N/A	1783

949655	62887	62902	TCATAATAGGCCGGGT	61	N/A	N/A	1784

949656	62969	62984	ATAATAACCACCGAAG	45	N/A	N/A	1785

949657	62972	62987	ATAATAATAACCACCG	13	N/A	N/A	1786

949658	63350	63365	CCTAGGATTTGTGTTA	101	N/A	N/A	1787

949659	63667	63682	GCCTAGGATTTATAGA	96	N/A	N/A	1788

949660	63759	63774	CCACAAATGAATGGTA	72	N/A	N/A	1789

949661	63780	63795	TGGGATGTGGTCTGGT	98	N/A	N/A	1790

949662	63826	63841	CTAATTACCAGCTAAC	46	N/A	N/A	1791

949663	63880	63895	ACTTAGGTTACTCCTA	76	N/A	N/A	1792

949664	64255	64270	AACTGCTGCTCCGATA	53	N/A	N/A	1793

949665	64313	64328	GGCTATAGGGCTGATA	104	N/A	N/A	1794

949666	64366	64381	AAATGCACCGCTCTGC	137	N/A	N/A	1795

949667	64530	64545	ACAGAATAAGACATCG	55	N/A	N/A	1796

949668	64650	64665	AGCAATCGCACATGCC	92	N/A	N/A	1797

949669	64984	64999	AGTAGATGCCAGAGGG	54	N/A	N/A	1798

949670	65026	65041	GGCTAAGCATGAAGGG	96	N/A	N/A	1799

949671	65086	65101	ATCTATCTCAGACATC	110	N/A	N/A	1800

949672	65109	65124	CCAAGGTCTGAACTTT	69	N/A	N/A	1801

949673	65111	65126	AACCAAGGTCTGAACT	125	N/A	N/A	1802

949674	65112	65127	CAACCAAGGTCTGAAC	110	N/A	N/A	1803

949675	65113	65128	GCAACCAAGGTCTGAA	31	N/A	N/A	1804

949676	65115	65130	TGGCAACCAAGGTCTG	67	N/A	N/A	1805

949677	65116	65131	CTGGCAACCAAGGTCT	120	N/A	N/A	1806

949678	65117	65132	GCTGGCAACCAAGGTC	156	N/A	N/A	1807

949679	65119	65134	ATGCTGGCAACCAAGG	40	N/A	N/A	1808

949680	65192	65207	GAATTTAGCCCTGACA	94	N/A	N/A	1809

949681	65208	65223	AGCTAAACAGGGTTTT	78	N/A	N/A	1810

949682	65343	65358	GTCCAAGGGCAGCCGG	100	N/A	N/A	1811

949683	65383	65398	CCCCAAAGGCCCTTAG	97	N/A	N/A	1812

949684	65458	65473	GTGGACATCAGTGAGT	53	N/A	N/A	1813

949685	65527	65542	CTACAAGTGAGCTGAG	6	N/A	N/A	1814

949686	65543	65558	CAAAACGGGAGCAGGC	83	N/A	N/A	1815

949687	65544	65559	ACAAAACGGGAGCAGG	51	N/A	N/A	1816

949688	65545	65560	CACAAAACGGGAGCAG	76	N/A	N/A	1817

949689	65583	65598	AAAAAGGGTCCAGTGG	91	N/A	N/A	1818

949690	65599	65614	TCATTAGTACGCCAGG	24	N/A	N/A	1819

949691	65686	65701	CCCAACTGTGCTTTCC	26	N/A	N/A	1820

949692	66037	66052	TCACACACGGATTTTT	73	N/A	N/A	1821

949693	66148	66163	TTCTGCATTGCAGTGG	26	N/A	N/A	1822

949694	66218	66233	GTAAAAAGCATCTGGC	83	N/A	N/A	1823

949695	66272	66287	GCCAGAGTCACCATCA	73	N/A	N/A	1824

949696	66327	66342	TGCCAAGGTGAGGTCA	91	N/A	N/A	1825

949697	66373	66388	CGCCAGATCTCTCTTG	104	N/A	N/A	1826

949698	66787	66802	TGCCTACCATTGCACT	86	N/A	N/A	1827

949699	66882	66897	CGAGAAGGAGGTAGAA	82	N/A	N/A	1828

949700	66890	66905	AACTACACCGAGAAGG	101	N/A	N/A	1829

949701	66895	66910	TCCAAAACTACACCGA	72	N/A	N/A	1830

949702	66973	66988	ATCAAACGAGTATATT	91	N/A	N/A	1831

949703	66974	66989	TATCAAACGAGTATAT	101	N/A	N/A	1832

949704	66987	67002	AGCTAAAATCCACTAT	110	N/A	N/A	1833

949705	66998	67013	CAATAGGGAAGAGCTA	57	N/A	N/A	1834

949706	66999	67014	GCAATAGGGAAGAGCT	117	N/A	N/A	1835

949707	67000	67015	AGCAATAGGGAAGAGC	75	N/A	N/A	1836

949708	67377	67392	ATCAAGACTCTGAATA	86	N/A	N/A	1837

949709	67669	67684	TTCAAAGATAGGTGAG	32	N/A	N/A	1838

949710	67762	67777	CATTAAGTATTTGGAG	77	N/A	N/A	1839

949711	67770	67785	AAGAGCAACATTAAGT	111	N/A	N/A	1840

949712	67773	67788	GCGAAGAGCAACATTA	120	N/A	N/A	1841

949713	67793	67808	TGCGAATGCACATGGC	138	N/A	N/A	1842

949714	67803	67818	ACACAGAGGGTGCGAA	153	N/A	N/A	1843

949715	67805	67820	CAACACAGAGGGTGCG	47	N/A	N/A	1844

949716	67823	67838	TAACATGTTACTCAGA	75	N/A	N/A	1845

949717	67870	67885	TCAAGTATAGAGATGG	46	N/A	N/A	1846

949718	68110	68125	AGTACCTGATTCAGCG	147	N/A	N/A	1847

949719	68180	68195	TGCCAGTGAGGATTGC	56	N/A	N/A	1848

949720	68210	68225	AAAACAAGTGCTGCTC	77	N/A	N/A	1849

949721	68491	68506	GAATCAAGTACATTTC	77	N/A	N/A	1850

949722	68541	68556	GTAATAGTAGCAGCAC	29	N/A	N/A	1851

949723	68542	68557	TGTAATAGTAGCAGCA	20	N/A	N/A	1852

949724	68610	68625	GTCTTCTGGGTAAATG	30	N/A	N/A	1853

949725	68624	68639	AGGCAATGCATACAGT	25	N/A	N/A	1854

949726	68992	69007	GAACGATAGGCCAGGC	90	N/A	N/A	1855

949727	68999	69014	AAGAAGAGAACGATAG	67	N/A	N/A	1856

949728	69029	69044	GTTTATGCACCATGTG	20	N/A	N/A	1857

949729	69102	69117	AAAACAGCGAGGAGGA	90	N/A	N/A	1858

949730	69108	69123	TGCTAGAAAACAGCGA	109	N/A	N/A	1859

949731	69206	69221	CAGGAACCTACTCTCG	86	N/A	N/A	1860

949732	69213	69228	ACAAGGACAGGAACCT	97	N/A	N/A	1861

949733	69215	69230	CTACAAGGACAGGAAC	107	N/A	N/A	1862

949734	69361	69376	CTAGACACTGACTTCC	89	N/A	N/A	1863

949735	69486	69501	GGCCAGCTGGACTTTT	74	N/A	N/A	1864

949736	69589	69604	TACTAGGGCACTTAAG	149	N/A	N/A	1865

949737	69591	69606	GATACTAGGGCACTTA	69	N/A	N/A	1866

949738	69692	69707	ACCAAGGGTCCCTGCC	113	N/A	N/A	1867

949739	69781	69796	TACAAAGTGTCTGGCG	53	N/A	N/A	1868

949740	69782	69797	ATACAAAGTGTCTGGC	41	N/A	N/A	1869

949741	70034	70049	GATTAGGAGACAAGCC	87	N/A	N/A	1870

949742	70467	70482	ATCCATTCTGGACAAC	62	N/A	N/A	1871

949743	70664	70679	CACTAGAGGATAGGCG	118	N/A	N/A	1872

949744	70759	70774	GTAAAGCAGAGTTTGG	25	N/A	N/A	1873

949745	70760	70775	TGTAAAGCAGAGTTTG	46	N/A	N/A	1874

949746	70766	70781	GCAAAGTGTAAAGCAG	53	N/A	N/A	1875

949747	70787	70802	CCATAAAATCAGCCTA	71	N/A	N/A	1876

949748	70897	70912	AAAGAACTCGGGAGGT	65	N/A	N/A	1877

949749	71020	71035	TTAAGATCAAGAGATC	115	N/A	N/A	1878

949750	71155	71170	TCAAGATGAGCTCCCC	65	N/A	N/A	1879

949751	71255	71270	GTCCATCCAGACCACG	119	N/A	N/A	1880

949752	71260	71275	GGACAGTCCATCCAGA	84	N/A	N/A	1881

949753	71369	71384	GGAGACGGCACGGAGC	81	N/A	N/A	1882

949754	71371	71386	ACGGAGACGGCACGGA	87	N/A	N/A	1883

949755	71473	71488	ACCAAGGGCCAACACC	70	N/A	N/A	1884

949756	71697	71712	ACATACTGCAGCAGGG	52	N/A	N/A	1885

949757	71797	71812	AATCAATGAGTGGACG	29	N/A	N/A	1886

949758	71827	71842	ACGCAGAAGGCTTGCA	87	N/A	N/A	1887

949759	71908	71923	CCATTCTGAGCACTCC	41	N/A	N/A	1888

949760	72021	72036	TCTAAAGGAGGCTGTT	82	N/A	N/A	1889

949761	72175	72190	AGCCTAATTTTGCAAT	133	N/A	N/A	1890

949762	72938	72953	GGCCAACACTCATCTT	128	N/A	N/A	1891

949763	73188	73203	TGATATGCAGTGACAC	75	N/A	N/A	1892

949764	73296	73311	TCCCACACGGCTCCTG	144	N/A	N/A	1893

949765	73415	73430	GCACAACCTGCTCTAT	80	N/A	N/A	1894

949766	73519	73534	GAGCACCTGAGTCAGT	91	N/A	N/A	1895

949767	73639	73654	ACCAACCAAGTCATGG	137	N/A	N/A	1896

949768	73653	73668	AGCTAAGCAGCTGCAC	133	N/A	N/A	1897

949769	73742	73757	TTACAGGCCAAAAGCC	88	N/A	N/A	1898

949770	73868	73883	GAGGAGGCGGTGTCCT	122	N/A	N/A	1899

949771	73954	73969	CATAAAAGCACTCTGG	87	N/A	N/A	1900

949772	73957	73972	GAGCATAAAAGCACTC	92	N/A	N/A	1901

949773	73970	73985	GTCAAGGCTGTCTGAG	56	N/A	N/A	1902

949774	74231	74246	GCTGAATCTTGTGTTT	47	N/A	N/A	1903

949775	74259	74274	CACAACAACACTTCCT	73	N/A	N/A	1904

949776	74351	74366	CTTTAGAGGTGCAGAA	53	N/A	N/A	1905

949777	74592	74607	ATCCACTGAAGCCTCT	50	N/A	N/A	1906

949778	74872	74887	CTTAACAAGTAGTATT	14	N/A	N/A	1907

949779	74874	74889	GGCTTAACAAGTAGTA	38	N/A	N/A	1908

949780	74962	74977	GGCAACTGGTTACAAA	58	N/A	N/A	1909

949781	74967	74982	TGCAAGGCAACTGGTT	14	N/A	N/A	1910

949782	75001	75016	CTCCATGGTGAGAGTG	73	N/A	N/A	1911

949783	75055	75070	GCTAAAGCAAGAAGGC	87	N/A	N/A	1912

949784	75104	75119	AGAAAGACCGCCCATG	97	N/A	N/A	1913

949785	75219	75234	GCGGAGACAGAAGGCT	121	N/A	N/A	1914

949786	75320	75335	AAGCATCGGTTAAGGC	142	N/A	N/A	1915

949787	75846	75861	CCCAAGTGGTGAGGCT	74	N/A	N/A	1916

949788	76007	76022	GTTCAGGCAGGCGGTT	53	N/A	N/A	1917

949789	76107	76122	AACAAGAGACCTTGTC	47	N/A	N/A	1918

949790	76211	76226	GATCAGGCTACTCAGG	116	N/A	N/A	1919

949791	76415	76430	TCATAGCCAATATAAC	42	N/A	N/A	1920

949792	76430	76445	TATCACAGGGTGTGCT	67	N/A	N/A	1921

949793	76487	76502	TCGGAATGTACTCTTA	23	N/A	N/A	1922

949794	76518	76533	GAATTCGGGTTGAATC	107	N/A	N/A	1923

949795	76636	76651	GACAAAGCACACCTGG	81	N/A	N/A	1924

949796	76637	76652	GGACAAAGCACACCTG	100	N/A	N/A	1925

949797	76731	76746	CACTAAACTAATGACA	98	N/A	N/A	1926

949798	76734	76749	TAGCACTAAACTAATG	88	N/A	N/A	1927

949799	76737	76752	GCATAGCACTAAACTA	83	N/A	N/A	1928

949800	76739	76754	AGGCATAGCACTAAAC	80	N/A	N/A	1929

949801	76744	76759	AGCAAAGGCATAGCAC	91	N/A	N/A	1930

949802	76764	76779	ACCAAACCAGGACCAA	57	N/A	N/A	1931

949803	77124	77139	GTTAATGCTTCACCTG	25	N/A	N/A	1932

949804	77125	77140	GGTTAATGCTTCACCT	104	N/A	N/A	1933

949805	77175	77190	GTAAAGGGTGTGCACA	10	N/A	N/A	1934

949806	77224	77239	AGGCAAGCACAAGCTG	115	N/A	N/A	1935

949807	77245	77260	CCTTCATACCCCAACT	97	N/A	N/A	1936

949808	77343	77358	TACTGAAGTACCTGGG	102	N/A	N/A	1937

949809	77421	77436	TCCTAAGCATTCCTAA	113	N/A	N/A	1938

949810	77450	77465	GTAGGATGATCTTGTT	27	N/A	N/A	1939

949811	77458	77473	GTAAAAAAGTAGGATG	68	N/A	N/A	1940

949812	77502	77517	CCTTAAAAAGAGTCTC	75	N/A	N/A	1941

949813	77561	77576	GCTCATCTAATGGACA	46	N/A	N/A	1942

949814	77634	77649	GGCGAGACAGAGTAAG	12	N/A	N/A	1943

949815	77894	77909	TATACTGGACACGGTG	96	N/A	N/A	1944

949816	77895	77910	ATATACTGGACACGGT	88	N/A	N/A	1945

949817	77897	77912	ATATATACTGGACACG	36	N/A	N/A	1946

949818	78035	78050	GTTTTATTGGCCGGGT	20	N/A	N/A	1947

949819	78144	78159	AGTCACTGATCCTAAA	74	N/A	N/A	1948

949820	78149	78164	CAAGAAGTCACTGATC	69	N/A	N/A	1949

949821	78222	78237	ACAAAATGGGTTGAGG	38	N/A	N/A	1950

949822	78223	78238	TACAAAATGGGTTGAG	50	N/A	N/A	1951

949823	78262	78277	GGACAGGACTTTGCCC	119	N/A	N/A	1952

949824	78348	78363	CCACAATTCAGGTGGC	122	N/A	N/A	1953

949825	78366	78381	GGAAAGGTAGCCACTT	64	N/A	N/A	1954

949826	78470	78485	GCCCACTTATTCCTCC	120	N/A	N/A	1955

949827	78561	78576	CCAAATTCAAGAGCTC	45	N/A	N/A	1956

949828	78611	78626	TACCATTTTTCAGCAG	25	N/A	N/A	1957

949829	78678	78693	TCAAAACGGAGGTTTC	20	N/A	N/A	1958

949830	78711	78726	GACCATCCTATGAGGA	106	N/A	N/A	1959

949831	78878	78893	GACCAGAGACGGGAGG	58	N/A	N/A	1960

949832	78949	78964	GCCTAGAGGTAGGAGC	64	N/A	N/A	1961

949833	78961	78976	CCACAAGTTCAGGCCT	107	N/A	N/A	1962

949834	79024	79039	TCAAAGCTGAGTGTCA	25	N/A	N/A	1963

949835	79143	79158	TGCTAGGGACATCTTC	17	N/A	N/A	1964

949836	79212	79227	GTATATATCAGCTCAG	30	N/A	N/A	1965

949837	79225	79240	ACATAAGGATGTTGTA	126	N/A	N/A	1966

949838	79243	79258	ATCCATGTTCAGTTTC	13	N/A	N/A	1967

949839	79330	79345	CGGAAAAAGAAGTCCC	154	N/A	N/A	1968

949840	79375	79390	TTCAAGCAAATCACAC	96	N/A	N/A	1969

949841	79794	79809	TCAGTATGCACCACCA	6	N/A	N/A	1970

949842	79870	79885	AAGCATGACATGACAC	16	N/A	N/A	1971

949843	80250	80265	CAGAATATTTAACTCG	26	N/A	N/A	1972

949844	80347	80362	GACAAACCAAGCACAT	107	N/A	N/A	1973

949845	80435	80450	GTGGAACTTGCCTGAC	86	N/A	N/A	1974

949846	80542	80557	TCCAAGAGCCCTCTTA	11	N/A	N/A	1975

949847	80662	80677	CCTATTTGATCAGGAG	137	N/A	N/A	1976

949848	80759	80774	TGATTCAGTGCCGCCC	7	N/A	N/A	1977

949849	80769	80784	CCCAAAACCCTGATTC	17	N/A	N/A	1978

949850	80805	80820	TCACGAGGAAGCCATG	54	N/A	N/A	1979

949851	80870	80885	GGCGATGGAAGGGCAG	57	N/A	N/A	1980

949852	80981	80996	TTTCATTGGCCCTGGC	62	N/A	N/A	1981

949853	80987	81002	CAGCAATTTCATTGGC	63	N/A	N/A	1982

949854	81187	81202	TACCTTACTGGTACAC	67	N/A	N/A	1983

949855	81200	81215	GCCGAGGGAGCAATAC	102	N/A	N/A	1984

949856	81201	81216	TGCCGAGGGAGCAATA	49	N/A	N/A	1985

949857	81288	81303	GAAAAGGCACCAGTGG	39	N/A	N/A	1986

949858	81306	81321	CACAAAGCGATATGGG	7	N/A	N/A	1987

949859	81411	81426	GTGCATCTGTCTGGCC	79	N/A	N/A	1988

949860	81445	81460	TGCTAGGCTTGGTATC	95	N/A	N/A	1989

949861	81530	81545	CAGCAGTTGACGCGGG	127	N/A	N/A	1990

949862	81635	81650	CCGGAGCTGAGGGACA	117	N/A	N/A	1991

949863	81738	81753	AAAAATGGGCTGCCCC	103	N/A	N/A	1992

949864	81740	81755	CCAAAAATGGGCTGCC	90	N/A	N/A	1993

949865	81752	81767	GACTAAGTGGATCCAA	2	N/A	N/A	1994

949866	81769	81784	GCGCATAGAGGCACTG	56	N/A	N/A	1995

949867	81845	81860	AACCAGGCCCGCGGCG	90	N/A	N/A	1996

949868	81909	81924	GCGCAAGCACAGACTG	109	N/A	N/A	1997

949869	81953	81968	GCAGAAATTCCCTGGC	92	N/A	N/A	1998

949870	82004	82019	AGCTTAAGCAGCAAAT	111	N/A	N/A	1999

949871	82041	82056	TTTGGACTGTTTTCCC	8	N/A	N/A	2000

949872	82043	82058	TGTTTGGACTGTTTTC	2	N/A	N/A	2001

949873	82044	82059	CTGTTTGGACTGTTTT	1	N/A	N/A	2002

949874	82045	82060	CCTGTTTGGACTGTTT	1	N/A	N/A	2003

949875	82047	82062	AACCTGTTTGGACTGT	4	N/A	N/A	2004

949876	82048	82063	CAACCTGTTTGGACTG	5	N/A	N/A	2005

949877	82050	82065	TCCAACCTGTTTGGAC	27	N/A	N/A	2006

949878	82052	82067	TTTCCAACCTGTTTGG	44	N/A	N/A	2007

949879	82053	82068	TTTTCCAACCTGTTTG	17	N/A	N/A	2008

949880	82054	82069	GTTTTCCAACCTGTTT	4	N/A	N/A	2009

949881	82084	82099	ATGTCCATGACACCTG	17	N/A	N/A	2010

949882	82085	82100	CATGTCCATGACACCT	39	N/A	N/A	2011

949883	82087	82102	TTCATGTCCATGACAC	30	N/A	N/A	2012

949884	82089	82104	CATTCATGTCCATGAC	13	N/A	N/A	2013

949885	82091	82106	AACATTCATGTCCATG	51	N/A	N/A	2014

949886	82154	82169	TACAGAAGGGCCACGG	6	N/A	N/A	2015

949887	82155	82170	TTACAGAAGGGCCACG	10	N/A	N/A	2016

949888	82259	82274	GATTAGTCCATGCATG	99	N/A	N/A	2017

949889	82308	82323	ACACACGCAGGACAGG	85	N/A	N/A	2018

949890	82361	82376	GGTCATGCGGAGGGCA	41	N/A	N/A	2019

949891	82509	82524	TGTTACCTTCTCGGCA	19	N/A	N/A	2020

949892	82529	82544	GCGGAGATGAGAACAA	13	N/A	N/A	2021

949893	82619	82634	CTGGTAGGCGGAAGGG	77	N/A	N/A	2022

949894	82632	82647	CTCTAAGTGACAACTG	18	N/A	N/A	2023

949895	82720	82735	AACAAAGGCTGACCGC	85	N/A	N/A	2024

949896	82827	82842	GATGATGTGTTTCCTG	1	N/A	N/A	2025

949897	82836	82851	GGCTATACAGATGATG	7	N/A	N/A	2026

949898	82948	82963	AACCAGACCTCATGCA	22	N/A	N/A	2027

949899	82953	82968	AACTTAACCAGACCTC	12	N/A	N/A	2028

949900	82994	83009	CGCCAAATGCCCCCAC	29	N/A	N/A	2029

949901	83049	83064	GATAGGAACTCCTTGC	48	N/A	N/A	2030

949902	83051	83066	TCGATAGGAACTCCTT	96	N/A	N/A	2031

949903	83124	83139	CTGAAGGGCCTCGCCA	119	N/A	N/A	2032

949904	83183	83198	TGCCATGCAGGGCTTC	74	N/A	N/A	2033

949905	83283	83298	GTGGACCGGAGACAGC	125	N/A	N/A	2034

949906	83399	83414	TAGAAAATGCGGAGGG	91	N/A	N/A	2035

949907	83424	83439	CGGGAAGCAACATGAA	77	N/A	N/A	2036

949908	83429	83444	TGGCACGGGAAGCAAC	87	N/A	N/A	2037

949909	83464	83479	GAACAAGCAGAGTGGT	51	N/A	N/A	2038

949910	83566	83581	GGCCAGTACCTGGTCC	103	N/A	N/A	2039

949911	83622	83637	GAACAGTACACTTGAC	90	N/A	N/A	2040

949912	83627	83642	GCACAGAACAGTACAC	100	N/A	N/A	2041

949942	83908	83923	GGTTGACTACAAAGGG	5	N/A	N/A	2042

949943	83909	83924	AGGTTGACTACAAAGG	3	N/A	N/A	2043

949944	83911	83926	AGAGGTTGACTACAAA	4	N/A	N/A	2044

949945	83912	83927	GAGAGGTTGACTACAA	2	N/A	N/A	2045

949946	83914	83929	TTGAGAGGTTGACTAC	66	N/A	N/A	2046

949947	83916	83931	TTTTGAGAGGTTGACT	21	N/A	N/A	2047

949948	83918	83933	GTTTTTGAGAGGTTGA	2	N/A	N/A	2048

949949	83919	83934	GGTTTTTGAGAGGTTG	3	N/A	N/A	2049

949950	83920	83935	GGGTTTTTGAGAGGTT	5	N/A	N/A	2050

949951	83922	83937	GTGGGTTTTTGAGAGG	17	N/A	N/A	2051

949952	83923	83938	TGTGGGTTTTTGAGAG	25	N/A	N/A	2052

949953	83924	83939	CTGTGGGTTTTTGAGA	38	N/A	N/A	2053

949954	83926	83941	GCCTGTGGGTTTTTGA	24	N/A	N/A	2054

949955	84164	84179	CCATAACTTACTGAAG	69	N/A	N/A	2055

949956	84276	84291	GTAGGATAGGGAGCGG	31	N/A	N/A	2056

949957	84379	84394	TCCCAAATGCAGACGC	38	N/A	N/A	2057

949958	84397	84412	ACAAACAACACGCTTG	88	N/A	N/A	2058

949959	84399	84414	CAACAAACAACACGCT	88	N/A	N/A	2059

949960	84687	84702	AATTAGGTGGGTAAAG	64	N/A	N/A	2060

949961	84860	84875	GCCTTTGCAAGAAAGC	94	N/A	N/A	2061

949962	85204	85219	GGTTTCAACAAACAGT	11	N/A	N/A	2062

949963	85243	85258	AACAAAGTGACATAGC	8	N/A	N/A	2063

949964	85260	85275	GGAGAGGGACCATTTT	94	N/A	N/A	2064

949965	85323	85338	AATCAAAACTGGGCGC	100	N/A	N/A	2065

949966	85335	85350	AGCAATTGGCCAAATC	55	N/A	N/A	2066

949967	85362	85377	GGCATAGGTGGACTGG	21	N/A	N/A	2067

949968	85379	85394	AGCTACAGCAGCTATG	83	N/A	N/A	2068

949969	85438	85453	CTTACCAACAGTAACA	67	N/A	N/A	2069

949970	85541	85556	GGCTGGTTTAGTAAGG	75	N/A	N/A	2070

949971	85608	85623	GGGCAAGGCACTCGGA	54	N/A	N/A	2071

949972	85618	85633	GAATAAAGTAGGGCAA	44	N/A	N/A	2072

949973	85681	85696	CGTCACTGGACTCCCC	75	N/A	N/A	2073

949974	85784	85799	CCAGGATGAAGTTGGT	94	N/A	N/A	2074

949975	85876	85891	ATCAAGCTAGAAAGGT	19	N/A	N/A	2075

949976	85880	85895	AGCTATCAAGCTAGAA	82	N/A	N/A	2076

949977	85884	85899	CTCAAGCTATCAAGCT	120	N/A	N/A	2077

949978	85946	85961	CCAATTATGAACACTA	7	N/A	N/A	2078

949979	85984	85999	GTTCAAAGTGTTGCAT	9	N/A	N/A	2079

949980	86329	86344	AGCCACTGCCTTGTAT	74	N/A	N/A	2080

949981	86547	86562	AATCATAGCCTCAAAC	85	N/A	N/A	2081

949982	86551	86566	GAACAATCATAGCCTC	14	N/A	N/A	2082

949983	86933	86948	AATAATGTGCAGTTGT	14	N/A	N/A	2083

949984	87041	87056	CCCCAAGGTCTGGAGT	104	N/A	N/A	2084

949985	87160	87175	TCCCAGACAGGAGAAC	80	N/A	N/A	2085

949986	87349	87364	ACCTACTTGGCAAACC	128	N/A	N/A	2086

949987	87359	87374	CTCTAAAAGTACCTAC	127	N/A	N/A	2087

949988	87456	87471	CGACAGAGACTGTCTG	130	N/A	N/A	2088

949989	87574	87589	CTAAACTAACCATGGG	79	N/A	N/A	2089

949990	87595	87610	GAACATAGCACTGCCC	3	N/A	N/A	2090

949991	87597	87612	CTGAACATAGCACTGC	3	N/A	N/A	2091

949992	87598	87613	TCTGAACATAGCACTG	9	N/A	N/A	2092

949993	87680	87695	GCACACTGGTCGCTAA	43	N/A	N/A	2093

949994	87782	87797	TTGAAGCTAGGGAGAC	85	N/A	N/A	2094

949995	87817	87832	ACGGAAGGTTCTGGTT	69	N/A	N/A	2095

949996	87880	87895	GCACACTGAATCGACA	23	N/A	N/A	2096

949997	87882	87897	ACGCACACTGAATCGA	43	N/A	N/A	2097

949998	87893	87908	TGCCTGAGACCACGCA	152	N/A	N/A	2098

949999	88002	88017	TACCAGCGGCACCACC	72	N/A	N/A	2099

950000	88014	88029	AGCGAGAGGGCATACC	30	N/A	N/A	2100

950001	88119	88134	CACTACACCTTAACGC	70	N/A	N/A	2101

950002	88226	88241	CTCTAGAGGCCCTCCC	98	N/A	N/A	2102

950003	88271	88286	CACCGATAAATGTTGT	27	N/A	N/A	2103

950004	88308	88323	CAGCACGGAACAGATC	51	N/A	N/A	2104

950005	88397	88412	AGGGACTTCGGCTCAT	120	N/A	N/A	2105

950006	88497	88512	GCAAACCAAAGATGGG	47	N/A	N/A	2106

950007	88619	88634	GGTAAGTGGGAAGGCC	73	N/A	N/A	2107

950008	88766	88781	CCCCGTAGCTCTTCAG	24	N/A	N/A	2108

950009	88878	88893	TTTAAGGACTCCACCT	86	N/A	N/A	2109

950010	88992	89007	TGGCATCACCTGACTT	99	N/A	N/A	2110

950011	89116	89131	ATCCAAAATCCTCCCC	43	N/A	N/A	2111

950012	89192	89207	TCAAATAGCAGCCCAC	38	N/A	N/A	2112

950013	89231	89246	CCTAAGGGCCTCCTGG	102	N/A	N/A	2113

950014	89335	89350	CACAAGAAGGATTTTC	2	N/A	N/A	2114

950015	89459	89474	CAGCAATCGACCCACC	31	N/A	N/A	2115

950016	89560	89575	GCTGAGATTACGCACC	38	N/A	N/A	2116

950017	89706	89721	CTTATCTGGGCTATTT	7	N/A	N/A	2117

950018	89719	89734	TATTATAGGGCTTCTT	20	N/A	N/A	2118

950019	89813	89828	GAAAGTAGGCTTCTGT	6	N/A	N/A	2119

950020	89966	89981	TGTTACTTATCCATCA	7	N/A	N/A	2120

950021	89980	89995	CAACATATATCTCCTG	8	N/A	N/A	2121

950022	90075	90090	ACAAATCCCACTATAT	58	N/A	N/A	2122

950023	90081	90096	GCATAGACAAATCCCA	3	N/A	N/A	2123

950024	90175	90190	CGACAGGGTGTGTTTC	6	N/A	N/A	2124

950025	90197	90212	AAATTTAGAGGACATC	38	N/A	N/A	2125

950026	90276	90291	AGCCATCCAGTCTTAG	38	N/A	N/A	2126

950027	90541	90556	CTCAAGTAGTTCCCCT	12	N/A	N/A	2127

950028	90642	90657	CACTAGAATCCCGGAG	18	N/A	N/A	2128

950029	90673	90688	ACTTACTAAAGCGCAG	24	N/A	N/A	2129

950030	90722	90737	AGTAAATGTATCTGGT	3	N/A	N/A	2130

950031	90754	90769	GCGCAAGGGAAACGGC	97	N/A	N/A	2131

950032	90804	90819	TGAGCAGCCACAGGTA	31	N/A	N/A	2132

950033	90806	90821	AGTGAGCAGCCACAGG	45	N/A	N/A	2133

950034	90807	90822	AAGTGAGCAGCCACAG	83	N/A	N/A	2134

950035	90808	90823	CAAGTGAGCAGCCACA	27	N/A	N/A	2135

950036	90810	90825	ACCAAGTGAGCAGCCA	14	N/A	N/A	2136

950037	90812	90827	GGACCAAGTGAGCAGC	54	N/A	N/A	2137

950038	90837	90852	GCGAATGTGGCCGTTC	64	N/A	N/A	2138

950039	90889	90904	GAACAGACAGGGTTCA	97	N/A	N/A	2139

950040	90978	90993	CGGGAGACTGCAGAGG	44	N/A	N/A	2140

950041	91083	91098	AGCAGATGGCCTGGTA	18	N/A	N/A	2141

950042	91189	91204	ACATAGGGCCTTCTGG	38	N/A	N/A	2142

950043	91226	91241	AGCCAAGAGTCAAGTT	36	N/A	N/A	2143

950044	91384	91399	CATGAGCCCACCACTC	120	N/A	N/A	2144

950045	91538	91553	CTCCTAACATGCTAGA	75	N/A	N/A	2145

950046	91825	91840	CGCTCAAGGGAAAAAT	25	N/A	N/A	2146

950047	91934	91949	TCCCAAGCTCCCCTGA	51	N/A	N/A	2147

950048	92034	92049	GATACAGCCGTTCATA	11	N/A	N/A	2148

950049	92059	92074	TAATAAGGTACTGAAG	54	N/A	N/A	2149

950050	92634	92649	ACGAAGCTAGGAGGCG	108	N/A	N/A	2150

950051	92635	92650	CACGAAGCTAGGAGGC	35	N/A	N/A	2151

950052	92814	92829	GCATGATAATCCCAGC	98	N/A	N/A	2152

950053	93110	93125	AGCCGCCCAGGCCAGA	96	N/A	N/A	2153

950054	93183	93198	GCAGAAATTTTGCGAA	35	N/A	N/A	2154

950055	93185	93200	AGGCAGAAATTTTGCG	48	N/A	N/A	2155

950056	93235	93250	GGTGGAGTCCCCGCCC	165	N/A	N/A	2156

950057	93248	93263	GTGCAAGGTACATGGT	11	N/A	N/A	2157

950058	93303	93318	CGCAAAAATAAATCAT	83	N/A	N/A	2158

950059	93304	93319	GCGCAAAAATAAATCA	105	N/A	N/A	2159

950060	93322	93337	ATGTATTACCTACGGC	7	N/A	N/A	2160

950061	93383	93398	CATATTCATGGTGTTA	10	N/A	N/A	2161

950062	93496	93511	AGAGATGGAAGGGACG	89	N/A	N/A	2162

950063	93586	93601	CCACACTAAGCATGTA	38	N/A	N/A	2163

950064	93600	93615	TCTAATGTTTGCTACC	7	N/A	N/A	2164

950065	93601	93616	CTCTAATGTTTGCTAC	4	N/A	N/A	2165

950066	93612	93627	ACCACTAGATTCTCTA	56	N/A	N/A	2166

950067	93648	93663	GAACAGGGAATATTAG	79	N/A	N/A	2167

950068	93744	93759	TTATACCCAGGGCAAC	37	N/A	N/A	2168

950069	93854	93869	TAAGTATTTGCCATCC	6	N/A	N/A	2169

950070	93908	93923	GCTTAAGCGAGATCTT	42	N/A	N/A	2170

950071	93921	93936	GAATTCTGGTCTGGCT	26	N/A	N/A	2171

950072	93958	93973	CATAGGAGATAGAAGG	4	N/A	N/A	2172

950073	93960	93975	TGCATAGGAGATAGAA	34	N/A	N/A	2173

950074	94019	94034	CACAAGAGGGCAAGAT	36	N/A	N/A	2174

950075	94050	94065	TGCTAGATCAACTAGC	7	N/A	N/A	2175

950076	94058	94073	GCAAAGAGTGCTAGAT	89	N/A	N/A	2176

950077	94060	94075	AGGCAAAGAGTGCTAG	78	N/A	N/A	2177

950078	94065	94080	GCACAAGGCAAAGAGT	89	N/A	N/A	2178

950079	94078	94093	TGCTAGAGGAGCGGCA	114	N/A	N/A	2179

950080	94153	94168	CTATAGAACAGGAGTG	15	N/A	N/A	2180

950081	94155	94170	TGCTATAGAACAGGAG	9	N/A	N/A	2181

950082	94164	94179	AGCCACATCTGCTATA	89	N/A	N/A	2182

950083	94176	94191	CCTTAATCACTTAGCC	7	N/A	N/A	2183

950084	94200	94215	TATCATATAGGGAGAA	16	N/A	N/A	2184

950085	94215	94230	GCATAGAGAAGAGGGT	6	N/A	N/A	2185

950086	94270	94285	GACAAGAGGCAGCATC	7	N/A	N/A	2186

950087	94333	94348	TCACAGGCCAAGGATA	81	N/A	N/A	2187

950088	94371	94386	TGGCAGGAAACGGTGC	42	N/A	N/A	2188

950089	94450	94465	GCAAATATGATTCATC	1	N/A	N/A	2189

950090	94481	94496	AAGGAGGTGTTACCCA	60	N/A	N/A	2190

950091	94581	94596	CTATGAGTAGCAGGCT	3	N/A	N/A	2191

950092	94652	94667	GTCCAAGTGTAAGAGC	12	N/A	N/A	2192

950093	94695	94710	GCAAAAGGGAGCCCCT	69	N/A	N/A	2193

950094	94696	94711	AGCAAAAGGGAGCCCC	88	N/A	N/A	2194

950095	94770	94785	TGCCAGAAGAGCCCCG	84	N/A	N/A	2195

950096	94791	94806	CTACAAGAAAGACCCT	30	N/A	N/A	2196

950097	94805	94820	TAGGATCGGCTCAGCT	18	N/A	N/A	2197

950098	94810	94825	TGGCATAGGATCGGCT	54	N/A	N/A	2198

950099	94920	94935	GAACACATGAGGAACC	24	N/A	N/A	2199

950100	94977	94992	GTACAAGGGAGAAGCA	100	N/A	N/A	2200

950101	95047	95062	GGCCACGCTCATGACG	97	N/A	N/A	2201

950102	95086	95101	GTAGAGATGGGAACCA	4	N/A	N/A	2202

950103	95155	95170	AAGCAGGGATCCTCTG	114	N/A	N/A	2203

950104	95189	95204	GCGGTATGGGTGGCAG	106	N/A	N/A	2204

950105	95227	95242	GGCTAGGGTGCACAGT	60	N/A	N/A	2205

950106	95278	95293	CTTAGAAGGAAGTAGG	100	N/A	N/A	2206

950107	95378	95393	CCGTTAGCCATCCACC	73	N/A	N/A	2207

950108	95416	95431	CCCTAGGGAGCTGCAA	112	N/A	N/A	2208

950109	95443	95458	CCACACGGGTTCAGAG	43	N/A	N/A	2209

950110	95496	95511	GCACACCAAGCCTTGG	63	N/A	N/A	2210

950111	95645	95660	AGGGATGTGCCCGCAC	87	N/A	N/A	2211

950112	95745	95760	TGACAACCAGGTGAGG	24	N/A	N/A	2212

950113	95854	95869	TGGGAATGCCTCCATT	95	N/A	N/A	2213

950114	95954	95969	AGCCCCTGTGAGGTCA	66	N/A	N/A	2214

950115	96079	96094	GTACAGACTTATCCCT	28	N/A	N/A	2215

950116	96081	96096	GGGTACAGACTTATCC	95	N/A	N/A	2216

950117	96178	96193	GGCTAAAGAGTACCCA	63	N/A	N/A	2217

950118	96181	96196	GGTGGCTAAAGAGTAC	23	N/A	N/A	2218

950119	96298	96313	GACTACCCCGACCCAC	71	N/A	N/A	2219

950120	96399	96414	GCACGGCACGGCAGGA	24	N/A	N/A	2220

950121	96523	96538	ACGCACAGGGCGGCCT	111	N/A	N/A	2221

950122	96527	96542	AGGCACGCACAGGGCG	83	N/A	N/A	2222

950123	96646	96661	CTAACGGCAGGCAGCC	71	N/A	N/A	2223

950124	96752	96767	CATCAGGGTTTCAGGC	11	N/A	N/A	2224

950125	96860	96875	GGCTAGGGTCAAAGGT	21	N/A	N/A	2225

950126	96977	96992	GCCCATGGGAGGAGTC	80	N/A	N/A	2226

950127	97078	97093	GAAACCAACATGAGGC	20	N/A	N/A	2227

950128	97149	97164	CGCAATAGAGACACCT	32	N/A	N/A	2228

950129	97221	97236	GAACCTCGCTCCGTGC	165	N/A	N/A	2229

950130	97453	97468	AGTGACTGAGTTCTGC	22	N/A	N/A	2230

950131	97809	97824	GGTTACGGTTGTTTTT	26	N/A	N/A	2231

950132	97855	97870	GTACAATGTAAGCCTT	3	N/A	N/A	2232

950133	97920	97935	TGCTAGTGACTCCATC	27	N/A	N/A	2233

950134	98046	98061	AGTGATGGAACCCCGG	48	N/A	N/A	2234

950135	98098	98113	CGCAGATAGAAAAACT	26	N/A	N/A	2235

950136	98099	98114	TCGCAGATAGAAAAAC	60	N/A	N/A	2236

950137	98150	98165	GCACACTGTGGCCAAG	70	N/A	N/A	2237

950138	98259	98274	AGTGAACGAGTCCCTC	84	N/A	N/A	2238

950139	98353	98368	TTCTACATAGGACCAT	13	N/A	N/A	2239

950140	98399	98414	AAACATCACAGGGACC	43	N/A	N/A	2240

950141	98508	98523	TCAGAAGAGAGGACCT	42	N/A	N/A	2241

950142	98643	98658	AGACACAGGGTAATGC	35	N/A	N/A	2242

950143	98866	98881	CAACAATGCAACAACG	15	N/A	N/A	2243

950144	99310	99325	CGCCAGATTAATTTTG	46	N/A	N/A	2244

950145	99477	99492	AACAATTGTCTGGTGT	85	N/A	N/A	2245

950146	99478	99493	CAACAATTGTCTGGTG	63	N/A	N/A	2246

950147	99481	99496	TAACAACAATTGTCTG	20	N/A	N/A	2247

950148	99493	99508	TAAGACTATAGTTAAC	63	N/A	N/A	2248

950149	99519	99534	CAACAGAAGAGTAGGG	26	N/A	N/A	2249

950150	99522	99537	GTTCAACAGAAGAGTA	25	N/A	N/A	2250

950151	99626	99641	GATAAATACTAGAGGC	45	N/A	N/A	2251

950152	99810	99825	GATCAGGGCCATAGAA	79	N/A	N/A	2252

950153	99823	99838	CAAGAAACGGGTTGAT	71	N/A	N/A	2253

950154	99830	99845	AATTAAGCAAGAAACG	114	N/A	N/A	2254

950155	99893	99908	CAACAAGCTATCACAC	92	N/A	N/A	2255

950156	99912	99927	CTGGACCGCATCAGAG	92	N/A	N/A	2256

950157	99924	99939	TTGAAAGCCAACCTGG	105	N/A	N/A	2257

950158	100015	100030	TAACACAGCCATGGCT	123	N/A	N/A	2258

950159	100020	100035	CCACATAACACAGCCA	28	N/A	N/A	2259

950160	100125	100140	TTGCAGCACAGGGTCA	56	N/A	N/A	2260

950161	100161	100176	GGCAACAGGAGACGCG	62	N/A	N/A	2261

950162	100227	100242	AGAGGCTACATGGTGA	60	N/A	N/A	2262

950163	100328	100343	GAAACAACCAATATCC	58	N/A	N/A	2263

950164	100399	100414	CACGAAGGGTTGACAC	101	N/A	N/A	2264

950165	100435	100450	GGCCAGAGTCCTGACT	98	N/A	N/A	2265

950166	100535	100550	GTTACCAGGACAAAAC	126	N/A	N/A	2266

950167	100670	100685	TGCCATCCCAGGGACC	105	N/A	N/A	2267

950168	100782	100797	CTACAGAGCATTCCCC	89	N/A	N/A	2268

950169	100811	100826	TGCCAGCAAGTCCCCA	80	N/A	N/A	2269

950170	100929	100944	GCCAAGAGGCCCGCCT	171	N/A	N/A	2270

950171	101038	101053	ACCCGAATAGCAAGCG	54	N/A	N/A	2271

950172	101155	101170	GAGCATGGGCCCGCAG	139	N/A	N/A	2272

950173	101255	101270	GATGATCCTTTCACCC	90	N/A	N/A	2273

950174	101362	101377	CACCACCCTGGGTTTA	132	N/A	N/A	2274

950175	101483	101498	GCAGACCACGCCTGGC	44	N/A	N/A	2275

950176	101603	101618	TGCAAGGCGGAAAGAG	95	N/A	N/A	2276

950177	101718	101733	GGCATTCCCACCCAGG	84	N/A	N/A	2277

950178	101808	101823	GAAAACTAGACTTGCA	31	N/A	N/A	2278

950179	101810	101825	CTGAAAACTAGACTTG	44	N/A	N/A	2279

950180	101850	101865	GAAAAACACGAAGGGC	86	N/A	N/A	2280

950181	101951	101966	TCATTACGGCCGGCGC	109	N/A	N/A	2281

950182	102059	102074	CTAACTTGGGTGGTGG	43	N/A	N/A	2282

950183	102159	102174	AGATATTGATGTCTTG	7	N/A	N/A	2283

950184	102264	102279	GCAGAGGGAGCCCCGA	90	N/A	N/A	2284

950185	102314	102329	TCGCATAGAAGATTCG	38	N/A	N/A	2285

950186	102365	102380	CGGCAGGTGAGTGTCA	30	N/A	N/A	2286

950187	102465	102480	GACTACCTGTTCAGGA	112	N/A	N/A	2287

950188	102585	102600	GAAGAGTGGTGGTTTC	80	N/A	N/A	2288

950189	102639	102654	TGCGAGCCGGCCCCAG	61	N/A	N/A	2289

950190	102716	102731	CACGATCCCAAACCCT	79	N/A	N/A	2290

950191	102830	102845	TGCCGGCACGCACGCG	135	N/A	N/A	2291

950192	102935	102950	CAGCAGGGCCAAGTAA	61	N/A	N/A	2292

950193	103102	103117	GGTAACACAGGGCCTG	119	N/A	N/A	2293

950207	103286	103301	GAGCATAGGAAGAGGG	27	N/A	N/A	2294

950208	103302	103317	GCAAAGAAGTAGGGCA	81	N/A	N/A	2295

950209	103323	103338	GATTAGCCTGCCAGGC	20	N/A	N/A	2296

950210	103425	103440	CCTCATCTATCCAAGG	99	N/A	N/A	2297

950211	103552	103567	GGCACATGGGCTCAGC	119	N/A	N/A	2298

950212	103672	103687	CTCCAAGTTCAGGAGC	119	N/A	N/A	2299

950213	103832	103847	CGCAAGCCACAGCAGG	101	N/A	N/A	2300

950214	103971	103986	ACTCACCCAGTGATGT	111	N/A	N/A	2301

950215	104100	104115	GTTCAGGAAGCACCAA	20	N/A	N/A	2302

950216	104276	104291	AGCCTTACTCCCTTTC	34	N/A	N/A	2303

950217	104326	104341	CGCTTTAGGACATCTT	63	N/A	N/A	2304

950218	104410	104425	TAGGAGTGAGTTACCT	102	N/A	N/A	2305

950219	104458	104473	CACGAAGTGAATGGAA	13	N/A	N/A	2306

950220	104486	104501	ACGCACAACACAACTG	63	N/A	N/A	2307

950221	104560	104575	TGCCACTGGACTGACT	69	N/A	N/A	2308

950222	104606	104621	GACCTAGGTGCCTGAC	27	N/A	N/A	2309

950223	104678	104693	ACTCTACCCGCACCCT	96	N/A	N/A	2310

950224	104783	104798	CTCCATGATGCTCCCT	63	N/A	N/A	2311

950225	104895	104910	GTGTAGCATGCGCCAC	124	N/A	N/A	2312

950226	104953	104968	GGGAAAAGAGTCACCC	113	N/A	N/A	2313

950227	105113	105128	CTTCACCCCGAGCCAT	90	N/A	N/A	2314

950228	105248	105263	CGGGAAGCATCCCTGG	114	N/A	N/A	2315

950229	105253	105268	CAGCACGGGAAGCATC	53	N/A	N/A	2316

950230	105366	105381	TGCAAGCACCTGAGCC	99	N/A	N/A	2317

950231	105479	105494	CCCTATGGGTTCTGGC	78	N/A	N/A	2318

950232	105561	105576	TCACAATTGGTGCCTT	16	N/A	N/A	2319

950233	105622	105637	CCCCACCTCGGCCAAT	59	N/A	N/A	2320

950234	105668	105683	CTAAATGGAGCTCTTG	64	N/A	N/A	2321

950235	105671	105686	GGCCTAAATGGAGCTC	79	N/A	N/A	2322

950236	105727	105742	CATCACACTCCTCGAT	96	N/A	N/A	2323

950237	105864	105879	ATGTACTGTGAGCTTC	12	N/A	N/A	2324

950238	106002	106017	GCTAATCCCTGACTCA	37	N/A	N/A	2325

950239	106118	106133	ACTCAGCTGGAGTTGG	63	N/A	N/A	2326

950240	106174	106189	GACAAATGGGTCTCGA	124	N/A	N/A	2327

950241	106213	106228	GCCTAGAAGAGCTATG	97	N/A	N/A	2328

950242	106218	106233	GGCAAGCCTAGAAGAG	95	N/A	N/A	2329

950243	106269	106284	AGGCACACGGCTCAGG	55	N/A	N/A	2330

950244	106276	106291	GTACAGGAGGCACACG	82	N/A	N/A	2331

950245	106340	106355	TACTACTCTGGGTTTC	53	N/A	N/A	2332

950246	106457	106472	GAACATGCAGGGCCTG	13	N/A	N/A	2333

950247	106838	106853	AAGAAGTAGAAGTCGG	36	N/A	N/A	2334

950248	106946	106961	GGATGAGGTTGTGTGG	82	N/A	N/A	2335

950249	107060	107075	ACGCACATTTTCCTGT	59	N/A	N/A	2336

950250	107115	107130	TGCTAAACAGTGCTTG	76	N/A	N/A	2337

950251	107478	107493	AAAGTTTAGGCATGGT	22	N/A	N/A	2338

950252	107673	107688	CGTCAATACTAAAATA	119	N/A	N/A	2339

950253	108115	108130	GCCCGAATGTGTTTTG	72	N/A	N/A	2340

950254	108434	108449	CACCAGAACTTTATTA	92	N/A	N/A	2341

950255	108584	108599	GGCAAGTGTCTGAGTT	17	N/A	N/A	2342

950256	108685	108700	TCTGACTGATCAGGAA	85	N/A	N/A	2343

950257	108748	108763	GCACAAGCTTGACCCT	92	N/A	N/A	2344

950258	108970	108985	GAAAATGACCTGTCCT	92	N/A	N/A	2345

950259	109080	109095	CCGTGAGGGAACTTTC	69	N/A	N/A	2346

950260	109124	109139	AAGCAAGTGGGACCCC	79	N/A	N/A	2347

950261	109129	109144	GAACAAAGCAAGTGGG	62	N/A	N/A	2348

950262	109183	109198	GGCCAAGGGTGGCCAT	143	N/A	N/A	2349

950263	109205	109220	CGGCAAGGCATTTTAA	88	N/A	N/A	2350

950264	109407	109422	TTTAAGCGATCTCCTG	31	N/A	N/A	2351

950265	109408	109423	GTTTAAGCGATCTCCT	39	N/A	N/A	2352

950266	109516	109531	ACCCAACCTTGGGACA	75	N/A	N/A	2353

950267	109576	109591	CCCAAAGGATCTGCCC	101	N/A	N/A	2354

950268	109649	109664	TGCCTGGGTATGTTTG	76	N/A	N/A	2355

950269	109910	109925	AGATAGGCCTGCTTGC	70	N/A	N/A	2356

950270	109914	109929	CACCAGATAGGCCTGC	74	N/A	N/A	2357

950271	110212	110227	ACTCACATTGTTGATC	92	N/A	N/A	2358

950272	110607	110622	CTCTAGGCTTACCACA	53	N/A	N/A	2359

950273	110885	110900	GCCTAGGGTGCCTGGC	105	N/A	N/A	2360

950274	110899	110914	TACAAACAGGTATGGC	20	N/A	N/A	2361

950275	110900	110915	CTACAAACAGGTATGG	46	N/A	N/A	2362

950276	111043	111058	TCAAATTTGTTGTTGC	14	N/A	N/A	2363

950277	111093	111108	CCAAAGAGGATGGCCG	66	N/A	N/A	2364

950278	111150	111165	AGTTAGTGGACTTGAT	37	N/A	N/A	2365

950279	111166	111181	CTGCAGTGTTCACTCC	64	N/A	N/A	2366

950280	111195	111210	GCCTAAGTTAATTCAG	88	N/A	N/A	2367

950281	111339	111354	GCAACCAGGGTAAACG	94	N/A	N/A	2368

950282	111471	111486	ACTGAAGAGGAGCCCA	57	N/A	N/A	2369

950283	111548	111563	GCCAAAGGCCTGATAC	96	N/A	N/A	2370

950284	111581	111596	GGTTTATAAAGCTTCC	35	N/A	N/A	2371

950285	111781	111796	TCTTACCTCGGTCTTC	66	N/A	N/A	2372

950286	111872	111887	TGAAAGAGGTTCCTGC	36	N/A	N/A	2373

950287	111910	111925	GTTGACATTCATATAC	68	N/A	N/A	2374

950288	112131	112146	TGAGAATGGGAGCAAC	39	N/A	N/A	2375

950289	112147	112162	GAACACTGATCACTAC	32	N/A	N/A	2376

950290	112174	112189	AAACATACCAGGACAC	39	N/A	N/A	2377

950291	112192	112207	TCCTATCGGTATATGA	139	N/A	N/A	2378

950292	112222	112237	TCATAGCAGAAGAACC	28	N/A	N/A	2379

950293	112260	112275	TAAACTTGCAGGGTCA	9	N/A	N/A	2380

950294	112267	112282	CTAAAAGTAAACTTGC	98	N/A	N/A	2381

950295	112360	112375	TCCCATCTTTGGCTGC	37	N/A	N/A	2382

950296	112902	112917	TCCTACTGGATACAGA	58	N/A	N/A	2383

950299	113293	113308	GCCAATCCTGTTATAT	97	N/A	N/A	2384

950300	113614	113629	GCAGAATAATCCTGTT	77	N/A	N/A	2385

950301	113616	113631	TGGCAGAATAATCCTG	69	N/A	N/A	2386

950302	113780	113795	GTTGACACCATGCTTG	14	N/A	N/A	2387

950303	113923	113938	CCCCAGACTTTTTTGA	101	N/A	N/A	2388

950304	114269	114284	AACCAGGCTTCCGGAC	83	N/A	N/A	2389

950305	114450	114465	AGCTGTAGGGCTGGCT	61	N/A	N/A	2390

950306	114637	114652	TCTGGCCACGCCTTGC	93	N/A	N/A	2391

950307	114737	114752	CCTTAAGGAACTCCAT	109	N/A	N/A	2392

950308	114843	114858	CCTAATATCTCTAGGG	82	N/A	N/A	2393

950309	115000	115015	GCGCAGTGGGATCCTC	178	N/A	N/A	2394

950310	115042	115057	TGCCAGAACCTGAGGT	96	N/A	N/A	2395

950311	115081	115096	ATACAGATGGAGTAGG	63	N/A	N/A	2396

950312	115423	115438	ACAGGAGGAGAGACCC	58	N/A	N/A	2397

950313	115497	115512	CCAAATAGGGATGAGG	87	N/A	N/A	2398

950314	115504	115519	TCGCAAGCCAAATAGG	29	N/A	N/A	2399

950315	115538	115553	GAACAAGCCTCTTGGC	142	N/A	N/A	2400

950316	115640	115655	GCCCAGTGTGAGGGTT	102	N/A	N/A	2401

950317	115795	115810	CATCAACTCGCCTGCT	101	N/A	N/A	2402

950318	115915	115930	CCGGGAGCAAGAGGCA	160	N/A	N/A	2403

951323	3692	3707	CCAGATGGCAGACTTC	19	N/A	N/A	2404

951324	3772	3787	AGATATCTAGATGGCC	111	N/A	N/A	2405

951325	3773	3788	TAGATATCTAGATGGC	63	N/A	N/A	2406

951326	3774	3789	ATAGATATCTAGATGG	52	N/A	N/A	2407

951327	3884	3899	CCAAATGGTGGTGGCA	84	N/A	N/A	2408

951328	4373	4388	CCAATTTGTAGGCTGT	9	N/A	N/A	2409

951329	4487	4502	ACAGATGGGCACACCG	91	N/A	N/A	2410

951330	4647	4662	ACAAATGAGTAGTTCC	6	N/A	N/A	2411

951331	4668	4683	TCATTTGGCAGCCACA	41	N/A	N/A	2412

951332	4670	4685	TATCATTTGGCAGCCA	77	N/A	N/A	2413

951333	4673	4688	TGTTATCATTTGGCAG	13	N/A	N/A	2414

951334	4767	4782	CCTTACTTGGGATTCA	64	N/A	N/A	2415

951335	4792	4807	ATGATAACACAGCCTC	47	N/A	N/A	2416

951336	4793	4808	GATGATAACACAGCCT	46	N/A	N/A	2417

951337	4795	4810	GGGATGATAACACAGC	104	N/A	N/A	2418

951338	4917	4932	GAGATTTGTGCTGCTT	16	N/A	N/A	2419

951339	5744	5759	GTAAGTAAGCACTTTT	45	N/A	N/A	2420

951340	5899	5914	GGATTATAGCAATGCC	12	N/A	N/A	2421

951341	5970	5985	TCACTTTACAGCCTCC	9	N/A	N/A	2422

951342	6124	6139	GGATTCTAGGAGGTCG	36	N/A	N/A	2423

951343	6458	6473	ATTAATACTCTTAACG	57	N/A	N/A	2424

951344	6738	6753	ACTAACTGGGCTGTTC	53	N/A	N/A	2425

951345	6740	6755	GAACTAACTGGGCTGT	40	N/A	N/A	2426

951346	6857	6872	GGAAATCTAACACTGG	7	N/A	N/A	2427

951347	6908	6923	AACTCTATAGATCTGA	101	N/A	N/A	2428

951348	6959	6974	AGACATGGTGGAACCA	81	N/A	N/A	2429

951349	7117	7132	GATTACAACAACAGCC	76	N/A	N/A	2430

951350	7175	7190	TGAAATCGAAGCCATT	95	N/A	N/A	2431

951351	7399	7414	TAAATGACAAGGGTCA	83	N/A	N/A	2432

951352	7942	7957	GCTATTATAACTTGTA	8	N/A	N/A	2433

951353	8568	8583	ATTTTAAAGAAGACGG	42	N/A	N/A	2434

951354	8583	8598	GCTATTATAACTCCTA	11	N/A	N/A	2435

951355	8616	8631	CACTTAATCCCTATTC	74	N/A	N/A	2436

951356	9095	9110	ATACTTATCGCCTTTT	5	N/A	N/A	2437

951357	9099	9114	CATCATACTTATCGCC	6	N/A	N/A	2438

951358	9144	9159	TCCAACTGTGATCTCT	8	N/A	N/A	2439

951359	10032	10047	CCAAGGAACACATCAC	25	N/A	N/A	2440

951360	10064	10079	ACTAATAAGATCTGTT	97	N/A	N/A	2441

951361	10131	10146	TGCAATTTGTCTTTGC	78	N/A	N/A	2442

951362	10136	10151	CAAGATGCAATTTGTC	3	N/A	N/A	2443

951363	10700	10715	CGAGATCACAAATGCT	41	N/A	N/A	2444

951364	11386	11401	AGTAATGATGAGTTCC	3	N/A	N/A	2445

951365	11416	11431	CACTGATAGGCACTCC	8	N/A	N/A	2446

951366	11553	11568	ATTACTTGTTTGCAGC	8	N/A	N/A	2447

951367	11556	11571	CCAATTACTTGTTTGC	8	N/A	N/A	2448

951368	11572	11587	TAAATAACTCTCACGG	25	N/A	N/A	2449

951369	11573	11588	GTAAATAACTCTCACG	86	N/A	N/A	2450

951370	11575	11590	GTGTAAATAACTCTCA	10	N/A	N/A	2451

951371	11922	11937	GTTGATAGTCGGTTGT	4	N/A	N/A	2452

951372	11960	11975	GGTCATTAAAGATTCT	4	N/A	N/A	2453

951373	11990	12005	GGTACCTAACAGGTCT	93	N/A	N/A	2454

951374	12062	12077	GTCTTTCCGGTCTTTT	4	N/A	N/A	2455

951375	12148	12163	GATTATAAACCTAATC	121	N/A	N/A	2456

951376	12215	12230	TCAATAAGTAACTCTG	11	N/A	N/A	2457

951377	12216	12231	GTCAATAAGTAACTCT	140	N/A	N/A	2458

951378	12237	12252	ACCTATCAAGGGCTGT	27	N/A	N/A	2459

951379	12244	12259	GTTATGAACCTATCAA	20	N/A	N/A	2460

951380	12245	12260	TGTTATGAACCTATCA	47	N/A	N/A	2461

951381	12850	12865	GTTTACATGATGTTCT	7	N/A	N/A	2462

951382	12860	12875	GTAAATTACGGTTTAC	122	N/A	N/A	2463

951383	12861	12876	AGTAAATTACGGTTTA	14	N/A	N/A	2464

951384	12890	12905	CTTAATTACAGCTTGT	6	N/A	N/A	2465

951385	12934	12949	TCATTATTGGCTTGGA	41	N/A	N/A	2466

951386	12935	12950	CTCATTATTGGCTTGG	4	N/A	N/A	2467

951387	12971	12986	CCTTTATTCAGATTGT	3	N/A	N/A	2468

951388	13136	13151	AGTCATCCAGCTCCCG	54	N/A	N/A	2469

951389	13167	13182	ATAAATGTCCAGCAGC	44	N/A	N/A	2470

951390	13169	13184	TCATAAATGTCCAGCA	63	N/A	N/A	2471

951391	13170	13185	ATCATAAATGTCCAGC	32	N/A	N/A	2472

951392	13392	13407	GCCATATGTGTGGCTC	65	N/A	N/A	2473

951393	13487	13502	TGTAATCTTGGCCCAG	83	N/A	N/A	2474

951394	13521	13536	TGCATATCCAAAGCCA	83	N/A	N/A	2475

951395	14442	14457	CCACATGTGAAGCAGC	21	N/A	N/A	2476

951396	14476	14491	GGCTATTCCACTGCAG	99	N/A	N/A	2477

951397	14694	14709	TCTGTAATGGTTTCCC	5	N/A	N/A	2478

951398	14808	14823	TGTACCTAAGTACAGA	97	N/A	N/A	2479

951399	14824	14839	TCTATAGGTACCAGCA	50	N/A	N/A	2480

951400	15508	15523	AGTGAAATTCACTCCA	47	N/A	N/A	2481

951401	15651	15666	CTCTTTATATAGATGG	58	N/A	N/A	2482

951402	16748	16763	AGAGATACATCAACCC	35	N/A	N/A	2483

951403	17836	17851	AAATATAGCAGGGACC	51	N/A	N/A	2484

951404	17839	17854	ACTAAATATAGCAGGG	30	N/A	N/A	2485

951405	17842	17857	GGCACTAAATATAGCA	71	N/A	N/A	2486

951406	19042	19057	TAGTATACCAAGGTTA	11	N/A	N/A	2487

951407	19043	19058	GTAGTATACCAAGGTT	6	N/A	N/A	2488

951408	19760	19775	CAGTATGTATACTTCC	2	N/A	N/A	2489

951409	19761	19776	TCAGTATGTATACTTC	86	N/A	N/A	2490

951410	20345	20360	TCCTGATAAGATTCAC	17	N/A	N/A	2491

951411	20392	20407	TCAGTTATCACTGTCA	31	N/A	N/A	2492

951412	20458	20473	ACTAGTAACTTTCCAG	37	N/A	N/A	2493

951413	20741	20756	GGATTATTACATGAGT	11	N/A	N/A	2494

951414	20743	20758	GAGGATTATTACATGA	29	N/A	N/A	2495

951415	20864	20879	GCCAACTAATTCCAGA	46	N/A	N/A	2496

951416	21512	21527	ATATATGTGGAGGCAA	10	N/A	N/A	2497

951417	21513	21528	TATATATGTGGAGGCA	104	N/A	N/A	2498

951418	21515	21530	AATATATATGTGGAGG	4	N/A	N/A	2499

951419	21654	21669	CACTATTGGCACACCA	115	N/A	N/A	2500

951420	21655	21670	ACACTATTGGCACACC	122	N/A	N/A	2501

951421	21656	21671	AACACTATTGGCACAC	111	N/A	N/A	2502

951422	21840	21855	TTCTTAACAGGCTCCC	41	N/A	N/A	2503

951423	22010	22025	TAAGTATGCGCTGCCA	31	N/A	N/A	2504

951424	22134	22149	AGGATATAGGGTCTCC	97	N/A	N/A	2505

951425	22135	22150	AAGGATATAGGGTCTC	37	N/A	N/A	2506

951426	22212	22227	GAACATCATGGAGCCT	33	N/A	N/A	2507

951427	23308	23323	TTTGATACTGATCTCA	18	N/A	N/A	2508

951428	24284	24299	GGTATTATGGTATGTG	2	N/A	N/A	2509

951429	24285	24300	AGGTATTATGGTATGT	2	N/A	N/A	2510

951430	24396	24411	TCATATGTATGGGTTT	4	N/A	N/A	2511

951431	24398	24413	CTTCATATGTATGGGT	10	N/A	N/A	2512

951432	25701	25716	ATAGATGTAAGCAACT	51	N/A	N/A	2513

951433	25709	25724	GGAAATTCATAGATGT	113	N/A	N/A	2514

951434	25740	25755	ACTTATGGATGCAGCT	52	N/A	N/A	2515

951435	25741	25756	AACTTATGGATGCAGC	39	N/A	N/A	2516

951436	26160	26175	AGCATTGGTATCAGGC	35	N/A	N/A	2517

951437	26168	26183	TGGATATCAGCATTGG	17	N/A	N/A	2518

951438	26173	26188	ATGGATGGATATCAGC	41	N/A	N/A	2519

951439	26180	26195	TTCAATCATGGATGGA	72	N/A	N/A	2520

951440	27070	27085	GGAATACTTGCAGCTA	41	N/A	N/A	2521

951441	27071	27086	AGGAATACTTGCAGCT	11	N/A	N/A	2522

951442	27224	27239	CAACATGGCACTCCCT	64	N/A	N/A	2523

951443	27231	27246	TCTGATGCAACATGGC	12	N/A	N/A	2524

951444	27569	27584	ATCATTGGGTGAAGAC	43	N/A	N/A	2525

951445	27762	27777	GGTCTATTGGCACACA	32	N/A	N/A	2526

951446	27776	27791	AGATTAAGTCATCCGG	90	N/A	N/A	2527

951447	27777	27792	GAGATTAAGTCATCCG	61	N/A	N/A	2528

951448	27778	27793	AGAGATTAAGTCATCC	73	N/A	N/A	2529

951449	28228	28243	GCAAATGCTAGCTGCA	142	N/A	N/A	2530

951450	29274	29289	CGCTCTAATGTCCTGA	11	N/A	N/A	2531

951451	29308	29323	TGGATATCCAGACCCA	97	N/A	N/A	2532

951452	29321	29336	GCAGATCCTGCCTTGG	92	N/A	N/A	2533

951453	29584	29599	TGAACTAGAGGCAGGC	29	N/A	N/A	2534

951454	29585	29600	GTGAACTAGAGGCAGG	19	N/A	N/A	2535

951455	29974	29989	TGTAACTTTCAGCAGT	25	N/A	N/A	2536

951456	29999	30014	TGTGATTGCAGGCCAG	13	N/A	N/A	2537

951457	30053	30068	TTTTATAGTCTCCAAC	140	N/A	N/A	2538

951458	30107	30122	TGTATGGACAGCAGGC	13	N/A	N/A	2539

951459	30108	30123	GTGTATGGACAGCAGG	10	N/A	N/A	2540

951460	30242	30257	ATCAATCTTGGGAGCT	39	N/A	N/A	2541

951461	30246	30261	CTAAATCAATCTTGGG	40	N/A	N/A	2542

951462	30348	30363	ATATTAATGCAGTGGT	4	N/A	N/A	2543

951463	30349	30364	CATATTAATGCAGTGG	14	N/A	N/A	2544

951464	30350	30365	ACATATTAATGCAGTG	66	N/A	N/A	2545

951465	30407	30422	AGAATATCAACACCTT	69	N/A	N/A	2546

951466	30408	30423	TAGAATATCAACACCT	61	N/A	N/A	2547

951467	30426	30441	TTATCTAACAGTCCCA	45	N/A	N/A	2548

951468	30429	30444	CATTTATCTAACAGTC	104	N/A	N/A	2549

951469	30509	30524	TAGTGATATGCTGCCT	22	N/A	N/A	2550

951470	30852	30867	CACAATCCTGGCTATC	56	N/A	N/A	2551

951471	30931	30946	CCAGACATTGCTGTCG	108	N/A	N/A	2552

951472	31367	31382	ACATTTGACGGTTTGA	15	N/A	N/A	2553

951473	32256	32271	GGGAATGATGCCACCT	110	N/A	N/A	2554

951474	32270	32285	CCAATCTGTTATGTGG	23	N/A	N/A	2555

951475	32336	32351	TGGAATCCAGTATTTC	103	N/A	N/A	2556

951476	32498	32513	AGGAATCCTGCAGCTC	108	N/A	N/A	2557

951477	32722	32737	TCCTATTAGAGTGGCA	49	N/A	N/A	2558

951478	32905	32920	AGCATTAACAGAGCAG	92	N/A	N/A	2559

951479	33017	33032	GGCTTTACTCCAGCCA	104	N/A	N/A	2560

951480	33518	33533	AACAATAGCCATTGGT	107	N/A	N/A	2561

951481	34302	34317	TGTTTATTTACCGTTT	3	N/A	N/A	2562

951482	35180	35195	AGTACTATTGCATCAT	28	N/A	N/A	2563

951483	36041	36056	TCACATACAGACCCCA	59	N/A	N/A	2564

951484	36073	36088	CTAAATGTCACTTAGC	40	N/A	N/A	2565

951485	36087	36102	GGAACTAACACATGCT	23	N/A	N/A	2566

951486	36362	36377	GAAATCTACCTACTCC	105	N/A	N/A	2567

951487	36363	36378	TGAAATCTACCTACTC	121	N/A	N/A	2568

951488	36617	36632	GGCACCAACATTCACC	96	N/A	N/A	2569

951489	36660	36675	TCCGATTTTGCTTCTA	10	N/A	N/A	2570

951490	36665	36680	ACTTATCCGATTTTGC	23	N/A	N/A	2571

951491	36809	36824	TGTTATTCCGAACAGT	43	N/A	N/A	2572

951492	36911	36926	TCTCATGGCGACTGGC	77	N/A	N/A	2573

951493	36948	36963	GGAATAAAGGCTCTCG	22	N/A	N/A	2574

951494	37009	37024	TTAGATTGTATGACCC	9	N/A	N/A	2575

951495	37194	37209	GGCTTTGTTAGATGCC	107	N/A	N/A	2576

951496	37246	37261	ATTCATAGTACCCCAC	62	N/A	N/A	2577

951497	37304	37319	GTATTTGTAGTGTGGA	110	N/A	N/A	2578

951498	37305	37320	GGTATTTGTAGTGTGG	10	N/A	N/A	2579

951499	37446	37461	GACAATGGTGATGCCC	62	N/A	N/A	2580

951500	37566	37581	ATCTTAGGCAGACCCT	42	N/A	N/A	2581

951501	37570	37585	ACACATCTTAGGCAGA	14	N/A	N/A	2582

951502	37729	37744	ATTAAAATGGGTCTGC	77	N/A	N/A	2583

951503	37750	37765	CATTTAACCATGACAG	31	N/A	N/A	2584

951504	37751	37766	CCATTTAACCATGACA	32	N/A	N/A	2585

951505	37971	37986	ACCTTAAGTTTTTTGG	118	N/A	N/A	2586

951506	38196	38211	GCTAATAGCTATTCAA	39	N/A	N/A	2587

951507	38225	38240	CGCATTACTGAGTTCA	12	N/A	N/A	2588

951508	38335	38350	GTGACTAAGACAACCA	40	N/A	N/A	2589

951509	38538	38553	AGACATGTAACTCCCT	39	N/A	N/A	2590

951510	38540	38555	CTAGACATGTAACTCC	72	N/A	N/A	2591

951511	38592	38607	GCCTATCCATGCTGCT	105	N/A	N/A	2592

951512	39123	39138	CAGAATTGTTACAACC	47	N/A	N/A	2593

951513	39140	39155	CCATTAAAATTTGCCC	30	N/A	N/A	2594

951514	39141	39156	TCCATTAAAATTTGCC	44	N/A	N/A	2595

951515	39233	39248	TTAACTATATCTAACT	73	N/A	N/A	2596

951516	39236	39251	CGATTAACTATATCTA	87	N/A	N/A	2597

951517	39547	39562	ATTTATGCGATTGGCC	88	N/A	N/A	2598

951518	39548	39563	GATTTATGCGATTGGC	17	N/A	N/A	2599

951519	39549	39564	AGATTTATGCGATTGG	11	N/A	N/A	2600

951520	39555	39570	TGTTTAAGATTTATGC	41	N/A	N/A	2601

951521	39748	39763	AAAGTAATTCACTGCG	81	N/A	N/A	2602

951522	40198	40213	ATGGACTACAGTCTGT	110	N/A	N/A	2603

951523	40275	40290	TGCAATCACAGACATC	93	N/A	N/A	2604

951524	41080	41095	GTATTAACCTCATGAA	93	N/A	N/A	2605

951525	41081	41096	GGTATTAACCTCATGA	68	N/A	N/A	2606

951526	41125	41140	GTCTTTCCGTCACAGT	40	N/A	N/A	2607

951527	41273	41288	GGATAATTGGAGAGGG	94	N/A	N/A	2608

951528	41274	41289	GGGATAATTGGAGAGG	68	N/A	N/A	2609

951529	42072	42087	GGGATTATGATGAGGA	3	N/A	N/A	2610

951530	42327	42342	GATGATGGGTTCTAGC	85	N/A	N/A	2611

951531	42355	42370	GGGAGATGTGCTCTTT	49	N/A	N/A	2612

951532	42381	42396	GTATTACCCAGTGGCT	62	N/A	N/A	2613

951533	42382	42397	TGTATTACCCAGTGGC	15	N/A	N/A	2614

951534	42387	42402	GGGAATGTATTACCCA	131	N/A	N/A	2615

951535	42501	42516	TACAATCATTCCCAGC	89	N/A	N/A	2616

951536	42506	42521	AGCACTACAATCATTC	82	N/A	N/A	2617

951537	42569	42584	TACTGTACAGCTGTGC	8	N/A	N/A	2618

951538	42578	42593	GGCAATTGGTACTGTA	121	N/A	N/A	2619

951539	42599	42614	GAAAATTTGTACAGGA	55	N/A	N/A	2620

951540	43029	43044	TACATTAAAGGCTCGA	44	N/A	N/A	2621

951541	43843	43858	GTACATATCCTGGCCG	32	N/A	N/A	2622

951542	43871	43886	TACTATTCGACAGAGC	31	N/A	N/A	2623

951543	43873	43888	ATTACTATTCGACAGA	36	N/A	N/A	2624

951544	43876	43891	TAAATTACTATTCGAC	45	N/A	N/A	2625

951545	43877	43892	GTAAATTACTATTCGA	63	N/A	N/A	2626

951546	43878	43893	TGTAAATTACTATTCG	19	N/A	N/A	2627

951547	44416	44431	GAGATCAACACATTTC	57	N/A	N/A	2628

951548	44701	44716	AGAAATTACTCTTGCG	116	N/A	N/A	2629

951549	44902	44917	TCCAATGACAGCCCAG	26	N/A	N/A	2630

951550	44914	44929	GTGAATTCAATGTCCA	11	N/A	N/A	2631

951551	44929	44944	ACACATGTGGAACACG	80	N/A	N/A	2632

951552	45353	45368	TATCTATAACACATGA	44	N/A	N/A	2633

951553	45356	45371	GGTTATCTATAACACA	23	N/A	N/A	2634

951554	45446	45461	GTCTTTATTGCATGGG	8	N/A	N/A	2635

951555	45569	45584	GCTATAAAACCAGCCA	101	N/A	N/A	2636

951556	45590	45605	AGCATTGGATTGAGCA	89	N/A	N/A	2637

951557	46280	46295	AGCAATGGTGGCCGGG	109	N/A	N/A	2638

951558	46328	46343	CTATATTCAACAGGCT	24	N/A	N/A	2639

951559	46329	46344	ACTATATTCAACAGGC	61	N/A	N/A	2640

951560	46335	46350	GGCCATACTATATTCA	102	N/A	N/A	2641

951561	46428	46443	GGGATATACGCCCACC	8	N/A	N/A	2642

951562	46590	46605	TATTTAAAAGGGCCGC	23	N/A	N/A	2643

951563	46591	46606	GTATTTAAAAGGGCCG	97	N/A	N/A	2644

951564	46861	46876	CGAAATAGCAGAGCTC	75	N/A	N/A	2645

951565	46961	46976	TCTATAGTGACTCTCA	9	N/A	N/A	2646

951566	46962	46977	TTCTATAGTGACTCTC	6	N/A	N/A	2647

951567	47624	47639	GTTCATCCGGGTCAGG	36	N/A	N/A	2648

951568	47926	47941	GCAACTATGCATCCAC	141	N/A	N/A	2649

951569	48813	48828	GAACATGGCAGCCAGT	82	N/A	N/A	2650

951570	48973	48988	TGCTTAATGCCTTGCC	82	N/A	N/A	2651

951571	49051	49066	CGCAATGTGTTCATTT	27	N/A	N/A	2652

951572	49192	49207	ACTTTAACAAGTCCCG	36	N/A	N/A	2653

951573	49193	49208	CACTTTAACAAGTCCC	23	N/A	N/A	2654

951574	49227	49242	ACAGATGAAGCTACCA	27	N/A	N/A	2655

951575	49443	49458	GGTTATGGTCTTAGGT	16	N/A	N/A	2656

951576	49674	49689	TCGATTTGATTCTGAG	84	N/A	N/A	2657

951577	50575	50590	GACAAATTGCCACTGG	54	N/A	N/A	2658

951578	50595	50610	GCAAATATCTCCCAGT	19	N/A	N/A	2659

951579	50845	50860	ACAATGACGAGGCTGG	25	N/A	N/A	2660

951580	50979	50994	TCAAATCCCATCAGGA	90	N/A	N/A	2661

951581	51714	51729	TGTACTTGCGGGTTTT	42	N/A	N/A	2662

951582	51790	51805	AGCAATCACGCTGCCA	99	N/A	N/A	2663

951583	51798	51813	AGTCATCCAGCAATCA	43	N/A	N/A	2664

951584	51810	51825	ATTAATCTGGGCAGTC	38	N/A	N/A	2665

951585	51811	51826	GATTAATCTGGGCAGT	67	N/A	N/A	2666

951586	51816	51831	TAAAGGATTAATCTGG	48	N/A	N/A	2667

951587	51977	51992	ATTTTGATAGCTCCAC	33	N/A	N/A	2668

951588	51979	51994	GCATTTTGATAGCTCC	17	N/A	N/A	2669

951589	52601	52616	GGTTTAAAAGCTTCCA	98	N/A	N/A	2670

951590	52938	52953	GTAAATGCTGAATGCC	34	N/A	N/A	2671

951591	53084	53099	GGATATGGACAGCAGG	17	N/A	N/A	2672

951592	53085	53100	AGGATATGGACAGCAG	15	N/A	N/A	2673

951593	53553	53568	TCACATGCCAGACTTT	100	N/A	N/A	2674

951594	53781	53796	CACACTCACGCATGTT	102	N/A	N/A	2675

951595	53852	53867	TTTACCTTGAATGTCC	46	N/A	N/A	2676

951596	54009	54024	GCAAGCTACTCTCTCA	88	N/A	N/A	2677

951597	54160	54175	TCAATACCTGCACAGG	42	N/A	N/A	2678

951598	54161	54176	GTCAATACCTGCACAG	80	N/A	N/A	2679

951599	54177	54192	TGTATGAGGACTCAGA	42	N/A	N/A	2680

951600	54189	54204	ATCATAACAGGCTGTA	28	N/A	N/A	2681

951601	54190	54205	CATCATAACAGGCTGT	94	N/A	N/A	2682

951602	54199	54214	TAAAATGCGCATCATA	51	N/A	N/A	2683

951603	54255	54270	GTAATGAGCTCAGTCT	45	N/A	N/A	2684

951604	55117	55132	GAAACTACGGCACTCT	121	N/A	N/A	2685

951605	55385	55400	GGTAATAAGTCACACT	103	N/A	N/A	2686

951606	55528	55543	GATTAATATCGGCTGA	44	N/A	N/A	2687

951607	55530	55545	TGGATTAATATCGGCT	24	N/A	N/A	2688

951608	55531	55546	ATGGATTAATATCGGC	9	N/A	N/A	2689

951609	56236	56251	GTTCATCACGGAAGGT	120	N/A	N/A	2690

951610	56274	56289	CCGAATCACAGATTGA	71	N/A	N/A	2691

951611	56324	56339	GTAATTGACAAGCAGT	16	N/A	N/A	2692

951612	56327	56342	TAAGTAATTGACAAGC	29	N/A	N/A	2693

951613	56387	56402	GTATATAAGACCCGGG	65	N/A	N/A	2694

951614	56388	56403	TGTATATAAGACCCGG	86	N/A	N/A	2695

951615	56389	56404	GTGTATATAAGACCCG	53	N/A	N/A	2696

951616	56390	56405	AGTGTATATAAGACCC	41	N/A	N/A	2697

951617	56459	56474	GACTATTCAAGAGGGC	70	N/A	N/A	2698

951618	56567	56582	TCTCATGGCACTCACT	42	N/A	N/A	2699

951619	56617	56632	ATTCATGGTCCACCAG	101	N/A	N/A	2700

951620	56685	56700	GGTGATGCTGCAGAGC	36	N/A	N/A	2701

951621	56725	56740	CAGGATACATCACCCA	128	N/A	N/A	2702

951622	56858	56873	TATTATGAATCCCCCT	90	N/A	N/A	2703

951623	56859	56874	ATATTATGAATCCCCC	100	N/A	N/A	2704

951624	56861	56876	ACATATTATGAATCCC	52	N/A	N/A	2705

951625	57375	57390	AGTAATGACCCAGGCT	74	N/A	N/A	2706

951626	57642	57657	AGTGATGGGTCCCCAC	94	N/A	N/A	2707

951627	57738	57753	GTGAATATCAACTGTT	41	N/A	N/A	2708

951628	57916	57931	GCAGATCTGGCTGCCT	71	N/A	N/A	2709

951629	58905	58920	AGGATATTTGGCTGGG	70	N/A	N/A	2710

951630	59003	59018	ACTTATCGCAAGGAAC	71	N/A	N/A	2711

951631	59075	59090	GCGATATACCTTTCTG	26	N/A	N/A	2712

951632	59076	59091	TGCGATATACCTTTCT	19	N/A	N/A	2713

951633	59174	59189	TGAATGACAGGTGTGT	124	N/A	N/A	2714

951634	59346	59361	TACAATCACAATTGCT	47	N/A	N/A	2715

951635	60402	60417	GCCAATACCAAAGAAC	96	N/A	N/A	2716

951636	60469	60484	TCTAAATCAGGACAGT	86	N/A	N/A	2717

951637	61124	61139	GCAACATGGGCACTGC	100	N/A	N/A	2718

951638	61190	61205	TAAGATCGCATCTGGG	93	N/A	N/A	2719

951639	61274	61289	AGCTATCTGGGTTAGG	105	N/A	N/A	2720

951640	61338	61353	GGAAATAGACGATCTC	68	N/A	N/A	2721

951641	61460	61475	CCAATTTGGGCATCAA	102	N/A	N/A	2722

951642	61461	61476	GCCAATTTGGGCATCA	102	N/A	N/A	2723

951643	61530	61545	GTTATATGTGTGGCAT	35	N/A	N/A	2724

951644	61531	61546	TGTTATATGTGTGGCA	40	N/A	N/A	2725

951645	61532	61547	GTGTTATATGTGTGGC	9	N/A	N/A	2726

951646	61680	61695	GCTAAAATTCCACTCT	96	N/A	N/A	2727

951647	61796	61811	CGTAAACATGATTCCT	20	N/A	N/A	2728

951648	61841	61856	GTTATTAAGGACAACC	120	N/A	N/A	2729

951649	61857	61872	CGTTATTGTCATTTGT	46	N/A	N/A	2730

951650	61943	61958	TCGAATGAAGATGGTC	30	N/A	N/A	2731

951651	62255	62270	AATGATAAGGACCCCA	63	N/A	N/A	2732

951652	62257	62272	GTAATGATAAGGACCC	66	N/A	N/A	2733

951653	62258	62273	AGTAATGATAAGGACC	80	N/A	N/A	2734

951654	62398	62413	GTTAATTGTGCACTTG	15	N/A	N/A	2735

951655	62399	62414	TGTTAATTGTGCACTT	80	N/A	N/A	2736

951656	62400	62415	GTGTTAATTGTGCACT	109	N/A	N/A	2737

951657	62526	62541	AGTTATAAACCCTGGG	11	N/A	N/A	2738

951658	63805	63820	ACAATAATTGGCCAAA	86	N/A	N/A	2739

951659	63829	63844	GATCTAATTACCAGCT	128	N/A	N/A	2740

951660	63881	63896	TACTTAGGTTACTCCT	49	N/A	N/A	2741

951661	64400	64415	GACAATGGGTCAACGC	67	N/A	N/A	2742

951662	65042	65057	TGCTCTAAATGACTGC	96	N/A	N/A	2743

951663	65090	65105	CCACATCTATCTCAGA	112	N/A	N/A	2744

951664	65141	65156	AATATCTCAGCAGTGC	54	N/A	N/A	2745

951665	65142	65157	CAATATCTCAGCAGTG	126	N/A	N/A	2746

951666	65354	65369	GGCCTAACAGTGTCCA	105	N/A	N/A	2747

951667	65601	65616	AGTCATTAGTACGCCA	46	N/A	N/A	2748

951668	66403	66418	GTCTTGTGGGATGTGA	66	N/A	N/A	2749

951669	66821	66836	TCAATACACACTGTTG	85	N/A	N/A	2750

951670	66828	66843	GGATAATTCAATACAC	41	N/A	N/A	2751

951671	67824	67839	ATAACATGTTACTCAG	71	N/A	N/A	2752

951672	67825	67840	CATAACATGTTACTCA	87	N/A	N/A	2753

951673	67827	67842	GTCATAACATGTTACT	76	N/A	N/A	2754

951674	67828	67843	TGTCATAACATGTTAC	102	N/A	N/A	2755

951675	67840	67855	GGCACTCACAGCTGTC	119	N/A	N/A	2756

951676	68164	68179	AGCTGTTGTGCAGTGC	73	N/A	N/A	2757

951677	68238	68253	GCTAAATGTGGACTGA	28	N/A	N/A	2758

951678	68623	68638	GGCAATGCATACAGTC	44	N/A	N/A	2759

951679	68650	68665	GTTATTAACCCCCTGT	44	N/A	N/A	2760

951680	69011	69026	TCAATTACTTACAAGA	62	N/A	N/A	2761

951681	69030	69045	TGTTTATGCACCATGT	57	N/A	N/A	2762

951682	69227	69242	GAATTATGCCTTCTAC	25	N/A	N/A	2763

951683	69652	69667	GTCTATGTGGGCTGCC	80	N/A	N/A	2764

951684	69735	69750	ATCTATGGCATGGAGC	82	N/A	N/A	2765

951685	69737	69752	ACATCTATGGCATGGA	75	N/A	N/A	2766

951686	71409	71424	ACAGATAAATCAAGCC	50	N/A	N/A	2767

951687	71450	71465	CACTATGCGGCCACCA	93	N/A	N/A	2768

951688	71699	71714	GTACATACTGCAGCAG	113	N/A	N/A	2769

951689	71700	71715	GGTACATACTGCAGCA	107	N/A	N/A	2770

951690	71857	71872	GCGAATGCCATGCTTA	82	N/A	N/A	2771

951691	71968	71983	GACATTTAACACACTA	7	N/A	N/A	2772

951692	73303	73318	GGCTTAATCCCACACG	97	N/A	N/A	2773

951693	73694	73709	GAATTTGGGATTGTGG	21	N/A	N/A	2774

951694	73828	73843	TCATGCATGGCACTGT	102	N/A	N/A	2775

951695	73956	73971	AGCATAAAAGCACTCT	74	N/A	N/A	2776

951696	74038	74053	GCTATGAATTTCCTCC	10	N/A	N/A	2777

951697	74039	74054	GGCTATGAATTTCCTC	115	N/A	N/A	2778

951698	74365	74380	TGCTATGACCCTCACT	82	N/A	N/A	2779

951699	75020	75035	ACAGATAACAGTGGCG	24	N/A	N/A	2780

951700	75136	75151	GTAATTCCCGTTCTCT	16	N/A	N/A	2781

951701	75145	75160	TCTATTACCGTAATTC	23	N/A	N/A	2782

951702	75247	75262	TACATGAACAGTCTGT	50	N/A	N/A	2783

951703	75327	75342	AACTTACAAGCATCGG	91	N/A	N/A	2784

951704	75329	75344	TGAACTTACAAGCATC	94	N/A	N/A	2785

951705	75417	75432	ATCAATTTTAAACACG	31	N/A	N/A	2786

951706	75765	75780	ACAAATACATGAGGTC	32	N/A	N/A	2787

951707	75894	75909	GTATTATTCCCAGCTA	77	N/A	N/A	2788

951708	75895	75910	TGTATTATTCCCAGCT	102	N/A	N/A	2789

951709	76435	76450	GAGAATATCACAGGGT	10	N/A	N/A	2790

951710	76448	76463	TGTAATCAGTCAAGAG	18	N/A	N/A	2791

951711	76550	76565	GACAATTGTGACCGGC	88	N/A	N/A	2792

951712	76568	76583	CACAATCAGAGCATTC	34	N/A	N/A	2793

951713	76593	76608	TGATTTACAGGCCCAA	37	N/A	N/A	2794

951714	76594	76609	GTGATTTACAGGCCCA	50	N/A	N/A	2795

951715	76725	76740	ACTAATGACAGACAAC	27	N/A	N/A	2796

951716	76728	76743	TAAACTAATGACAGAC	92	N/A	N/A	2797

951717	76775	76790	TTGAATTGTGGACCAA	60	N/A	N/A	2798

951718	77126	77141	AGGTTAATGCTTCACC	54	N/A	N/A	2799

951719	77303	77318	TCAAATGGTGACCACT	112	N/A	N/A	2800

951720	77432	77447	CGTTTTAGTTTTCCTA	14	N/A	N/A	2801

951721	77538	77553	GTAATAACATGTGTCG	45	N/A	N/A	2802

951722	77539	77554	GGTAATAACATGTGTC	27	N/A	N/A	2803

951723	77570	77585	TAAACAATTGCTCATC	59	N/A	N/A	2804

951724	78076	78091	GGTAATTCTGCAAAGA	29	N/A	N/A	2805

951725	78080	78095	TAATGGTAATTCTGCA	43	N/A	N/A	2806

951726	78082	78097	ACTAATGGTAATTCTG	50	N/A	N/A	2807

951727	78166	78181	TTAGATCTCATTACCT	71	N/A	N/A	2808

951728	78206	78221	GTAACAACATCTACCT	88	N/A	N/A	2809

951729	78207	78222	GGTAACAACATCTACC	3	N/A	N/A	2810

951730	78387	78402	TGATTTACAGGATGCT	15	N/A	N/A	2811

951731	78388	78403	TTGATTTACAGGATGC	8	N/A	N/A	2812

951732	78539	78554	GCAAATGCTGGCTACG	85	N/A	N/A	2813

951733	78546	78561	CCCAATGGCAAATGCT	66	N/A	N/A	2814

951734	78848	78863	GGCACTACCTTCACCA	109	N/A	N/A	2815

951735	79046	79061	TCATATTGTTCTGCCC	15	N/A	N/A	2816

951736	79047	79062	TTCATATTGTTCTGCC	11	N/A	N/A	2817

951737	79077	79092	GGCTTTTGTCAGAGGA	8	N/A	N/A	2818

951738	79153	79168	TTCAATGACATGCTAG	19	N/A	N/A	2819

951739	79209	79224	TATATCAGCTCAGTGC	75	N/A	N/A	2820

951740	79211	79226	TATATATCAGCTCAGT	66	N/A	N/A	2821

951741	79214	79229	TTGTATATATCAGCTC	37	N/A	N/A	2822

951742	79229	79244	TCATACATAAGGATGT	91	N/A	N/A	2823

951743	79398	79413	GCTCATTGTAAACAGT	20	N/A	N/A	2824

951744	79462	79477	TGCATTTGGTCTTGCT	118	N/A	N/A	2825

951745	79799	79814	GACTATCAGTATGCAC	118	N/A	N/A	2826

951746	79845	79860	CATCATATTGGTGGGA	11	N/A	N/A	2827

951747	80265	80280	GCTATATTCACAAGGC	70	N/A	N/A	2828

951748	80266	80281	GGCTATATTCACAAGG	39	N/A	N/A	2829

951749	80520	80535	AACGATTAGGGACTCA	4	N/A	N/A	2830

951750	80663	80678	CCCTATTTGATCAGGA	117	N/A	N/A	2831

951751	80845	80860	GCAACTACCTTTGTTA	17	N/A	N/A	2832

951752	80986	81001	AGCAATTTCATTGGCC	105	N/A	N/A	2833

951753	81192	81207	AGCAATACCTTACTGG	46	N/A	N/A	2834

951754	81436	81451	TGGTATCAGGGTCCCA	130	N/A	N/A	2835

951755	81663	81678	CATGATGTCGCTGGCA	41	N/A	N/A	2836

951756	81741	81756	TCCAAAAATGGGCTGC	28	N/A	N/A	2837

951757	82016	82031	TCTAATCGAAGAAGCT	99	N/A	N/A	2838

951758	82262	82277	TCTGATTAGTCCATGC	40	N/A	N/A	2839

951759	82535	82550	ACAAATGCGGAGATGA	41	N/A	N/A	2840

951761	82653	82668	GCCAATGCAAGGTGGC	119	N/A	N/A	2841

951762	82803	82818	TGTTTTGGTGCTGGCT	99	N/A	N/A	2842

951763	82880	82895	TGGAATGGTTACTCTC	3	N/A	N/A	2843

951764	82920	82935	ATCTTCTGCAGTGTGG	2	N/A	N/A	2844

951765	83070	83085	GGTATCTGGGCCATCA	101	N/A	N/A	2845

951766	83419	83434	AGCAACATGAACCACA	26	N/A	N/A	2846

951767	84425	84440	GTAATCAAGGCAGCAA	7	N/A	N/A	2847

951768	85210	85225	AGCTCTGGTTTCAACA	82	N/A	N/A	2848

951769	85217	85232	TTAATTAAGCTCTGGT	12	N/A	N/A	2849

951770	85218	85233	CTTAATTAAGCTCTGG	22	N/A	N/A	2850

951771	85220	85235	GGCTTAATTAAGCTCT	67	N/A	N/A	2851

951772	85238	85253	AGTGACATAGCAAGCA	7	N/A	N/A	2852

951773	85417	85432	AAGAATCAGACTCTTC	110	N/A	N/A	2853

951774	85726	85741	CATTTATAACAGGCCA	82	N/A	N/A	2854

951775	85727	85742	CCATTTATAACAGGCC	94	N/A	N/A	2855

951776	85844	85859	GGCTTATGAGTCCACT	91	N/A	N/A	2856

951777	85973	85988	TGCATTAAAACTGCGA	87	N/A	N/A	2857

951778	86550	86565	AACAATCATAGCCTCA	114	N/A	N/A	2858

951779	86582	86597	AACTTACTGTCTCCCA	37	N/A	N/A	2859

951780	86937	86952	GTTTAATAATGTGCAG	40	N/A	N/A	2860

951781	86938	86953	GGTTTAATAATGTGCA	11	N/A	N/A	2861

951782	86939	86954	GGGTTTAATAATGTGC	27	N/A	N/A	2862

951783	87671	87686	TCGCTAAAGGCTCTGA	10	N/A	N/A	2863

951784	87858	87873	TGAAATCAGGGCTCCC	31	N/A	N/A	2864

951785	88030	88045	CTCTATCACAGGGCCC	92	N/A	N/A	2865

951786	88032	88047	TGCTCTATCACAGGGC	30	N/A	N/A	2866

951787	88258	88273	TGTATCTGGTGACTCC	5	N/A	N/A	2867

951788	88270	88285	ACCGATAAATGTTGTA	65	N/A	N/A	2868

951789	88634	88649	TATAGTAACAGCTGGG	73	N/A	N/A	2869

951790	88637	88652	CAGTATAGTAACAGCT	94	N/A	N/A	2870

951791	88638	88653	TCAGTATAGTAACAGC	41	N/A	N/A	2871

951792	88880	88895	TGTTTAAGGACTCCAC	28	N/A	N/A	2872

951793	88887	88902	GACAATTTGTTTAAGG	5	N/A	N/A	2873

951794	89152	89167	GATCTAATCAGAAGCA	29	N/A	N/A	2874

951795	89558	89573	TGAGATTACGCACCAC	5	N/A	N/A	2875

951796	89718	89733	ATTATAGGGCTTCTTA	30	N/A	N/A	2876

951797	89720	89735	TTATTATAGGGCTTCT	8	N/A	N/A	2877

951798	89721	89736	TTTATTATAGGGCTTC	1	N/A	N/A	2878

951799	89722	89737	GTTTATTATAGGGCTT	1	N/A	N/A	2879

951800	89746	89761	AGTGATATTCCCAACA	19	N/A	N/A	2880

951801	89977	89992	CATATATCTCCTGTTA	20	N/A	N/A	2881

951802	89999	90014	TCATATCACCACTTGC	29	N/A	N/A	2882

951803	90003	90018	ACAATCATATCACCAC	6	N/A	N/A	2883

951804	90004	90019	GACAATCATATCACCA	3	N/A	N/A	2884

951805	90058	90073	TTATATGTGGACACCT	34	N/A	N/A	2885

951806	90059	90074	TTTATATGTGGACACC	10	N/A	N/A	2886

951807	90139	90154	CACAATCCCAGATCTC	7	N/A	N/A	2887

951808	90161	90176	TCTATAAGGTTCCTTA	14	N/A	N/A	2888

951809	90165	90180	TGTTTCTATAAGGTTC	3	N/A	N/A	2889

951810	90199	90214	CCAAATTTAGAGGACA	46	N/A	N/A	2890

951811	90222	90237	ACATATCTTGCATGAC	4	N/A	N/A	2891

951812	90264	90279	TTAGACATCAGACTGT	76	N/A	N/A	2892

951813	90267	90282	GTCTTAGACATCAGAC	115	N/A	N/A	2893

951814	90706	90721	TCAATTGAGGCTTCTA	7	N/A	N/A	2894

951815	90716	90731	TGTATCTGGTTCAATT	11	N/A	N/A	2895

951816	90721	90736	GTAAATGTATCTGGTT	2	N/A	N/A	2896

951817	91071	91086	GGTACTGAGGCTGTGG	28	N/A	N/A	2897

951818	91117	91132	CAGAATTATCAATTCC	145	N/A	N/A	2898

951819	91174	91189	GGTGGGTAAAATGTGC	63	N/A	N/A	2899

951820	91191	91206	GCACATAGGGCCTTCT	79	N/A	N/A	2900

951821	91909	91924	GACAATGTAGTTTCCA	2	N/A	N/A	2901

951822	91939	91954	AACTATCCCAAGCTCC	10	N/A	N/A	2902

951823	91945	91960	GGCTTCAACTATCCCA	39	N/A	N/A	2903

951824	92037	92052	ATTGATACAGCCGTTC	8	N/A	N/A	2904

951825	92827	92842	ATGATAAAGGCTGGCA	21	N/A	N/A	2905

951826	92828	92843	AATGATAAAGGCTGGC	127	N/A	N/A	2906

951827	93169	93184	AAGATTATGGTTTCCT	4	N/A	N/A	2907

951828	93170	93185	GAAGATTATGGTTTCC	6	N/A	N/A	2908

951829	93241	93256	GTACATGGTGGAGTCC	83	N/A	N/A	2909

951830	93321	93336	TGTATTACCTACGGCA	6	N/A	N/A	2910

951831	93392	93407	GCTATTAATCATATTC	18	N/A	N/A	2911

951832	93572	93587	TACTTACCAGGATCTC	24	N/A	N/A	2912

951833	93746	93761	ACTTATACCCAGGGCA	7	N/A	N/A	2913

951834	93747	93762	GACTTATACCCAGGGC	81	N/A	N/A	2914

951835	93923	93938	ATGAATTCTGGTCTGG	2	N/A	N/A	2915

951836	94043	94058	TCAACTAGCCATGTCT	25	N/A	N/A	2916

951837	94169	94184	CACTTAGCCACATCTG	5	N/A	N/A	2917

951838	94174	94189	TTAATCACTTAGCCAC	8	N/A	N/A	2918

951839	94177	94192	ACCTTAATCACTTAGC	5	N/A	N/A	2919

951840	94182	94197	ACAATACCTTAATCAC	7	N/A	N/A	2920

951841	94284	94299	CAATTTATCTGGAGGA	8	N/A	N/A	2921

951842	94350	94365	TCAATTAGTGTTGCCT	119	N/A	N/A	2922

951843	94351	94366	ATCAATTAGTGTTGCC	5	N/A	N/A	2923

951844	94423	94438	GTCAATCGACTGGTAC	41	N/A	N/A	2924

951845	94446	94461	ATATGATTCATCTCGG	2	N/A	N/A	2925

951846	94447	94462	AATATGATTCATCTCG	2	N/A	N/A	2926

951847	94583	94598	AGCTATGAGTAGCAGG	18	N/A	N/A	2927

951848	94885	94900	GCCAACTAATTCCTCC	31	N/A	N/A	2928

951849	96000	96015	GCCATTCACGGGCTGC	104	N/A	N/A	2929

951850	96084	96099	CAAGGGTACAGACTTA	152	N/A	N/A	2930

951851	96224	96239	GGCTGGATGTGAGTGC	63	N/A	N/A	2931

951852	96650	96665	ACAACTAACGGCAGGC	17	N/A	N/A	2932

951853	96755	96770	ACACATCAGGGTTTCA	56	N/A	N/A	2933

951854	97483	97498	TCCTAATGGTGACTCA	40	N/A	N/A	2934

951855	97854	97869	TACAATGTAAGCCTTT	7	N/A	N/A	2935

951856	98522	98537	GTATTTCAAAGTAGTC	4	N/A	N/A	2936

951857	98652	98667	GGTTTTAATAGACACA	6	N/A	N/A	2937

951858	98653	98668	GGGTTTTAATAGACAC	53	N/A	N/A	2938

951859	99483	99498	GTTAACAACAATTGTC	60	N/A	N/A	2939

951860	99712	99727	ACAGATCTTGCATGTT	84	N/A	N/A	2940

951861	99888	99903	AGCTATCACACAGAAG	95	N/A	N/A	2941

951862	100033	100048	GCAAATCCATTGCCCA	78	N/A	N/A	2942

951863	100171	100186	GTGTTAAGGTGGCAAC	43	N/A	N/A	2943

951864	100222	100237	CTACATGGTGATGGGT	20	N/A	N/A	2944

951865	100285	100300	GTTAGCTGGACTCACA	89	N/A	N/A	2945

951866	100375	100390	CAATATCCTGGTGACT	36	N/A	N/A	2946

951867	100514	100529	ACACATGCAGTGAGCC	60	N/A	N/A	2947

951868	101384	101399	TGTATGGTGGCTCAGC	26	N/A	N/A	2948

951869	101385	101400	TTGTATGGTGGCTCAG	44	N/A	N/A	2949

951870	101796	101811	TGCAATGCTGCCAGGC	72	N/A	N/A	2950

951871	102161	102176	AGAGATATTGATGTCT	51	N/A	N/A	2951

951872	102425	102440	GCACATTGGTTAGGCT	93	N/A	N/A	2952

951873	102445	102460	CTCAATGTGTACAGTA	22	N/A	N/A	2953

951874	102885	102900	TGGAACTGTTTACAGT	10	N/A	N/A	2954

951875	102926	102941	CAAGTAACTGAGTCAC	7	N/A	N/A	2955

951876	103421	103436	ATCTATCCAAGGAGGA	93	N/A	N/A	2956

951877	103897	103912	TCAGTTCACGGATCCC	42	N/A	N/A	2957

951878	104120	104135	TGCTTTCTAAGGTTGG	14	N/A	N/A	2958

951879	104441	104456	TTTTATCTGGGCTCCA	39	N/A	N/A	2959

951880	104447	104462	TGGAACTTTTATCTGG	17	N/A	N/A	2960

951881	104452	104467	GTGAATGGAACTTTTA	5	N/A	N/A	2961

951882	104843	104858	TGTATATGAGCCCATC	99	N/A	N/A	2962

951883	105539	105554	TCATATCCATGCCCAC	31	N/A	N/A	2963

951884	105559	105574	ACAATTGGTGCCTTTC	14	N/A	N/A	2964

951885	105560	105575	CACAATTGGTGCCTTT	20	N/A	N/A	2965

951886	105730	105745	GGACATCACACTCCTC	100	N/A	N/A	2966

951887	105777	105792	CGCAATGGTGACTGCC	114	N/A	N/A	2967

951888	105791	105806	TGAAATACCATCCACG	40	N/A	N/A	2968

951889	105793	105808	CTTGAAATACCATCCA	19	N/A	N/A	2969

951890	106015	106030	GCCTTTACTGACAGCT	71	N/A	N/A	2970

951891	106173	106188	ACAAATGGGTCTCGAG	101	N/A	N/A	2971

951892	106195	106210	GGTTTTCACGGCCAGC	44	N/A	N/A	2972

951893	106203	106218	GCTATGAAGGTTTTCA	10	N/A	N/A	2973

951894	106204	106219	AGCTATGAAGGTTTTC	20	N/A	N/A	2974

951895	106233	106248	TGTCATATGGCCAGGG	17	N/A	N/A	2975

951896	106295	106310	TAATATTGTCACCCTC	93	N/A	N/A	2976

951897	106296	106311	ATAATATTGTCACCCT	29	N/A	N/A	2977

951898	106351	106366	GTTTTTGCAGCTACTA	9	N/A	N/A	2978

951899	106371	106386	ACTATCAACCATCTTA	20	N/A	N/A	2979

951900	106391	106406	GGACTTAATTTACACA	12	N/A	N/A	2980

951901	106392	106407	TGGACTTAATTTACAC	30	N/A	N/A	2981

951902	106404	106419	GATACTAAAGTCTGGA	11	N/A	N/A	2982

951903	107100	107115	GCAGATGGTGGTCTAC	69	N/A	N/A	2983

951904	109198	109213	GCATTTTAATATTGGG	31	N/A	N/A	2984

951905	109409	109424	GGTTTAAGCGATCTCC	46	N/A	N/A	2985

951906	109955	109970	CCAATAACTGACCTCA	72	N/A	N/A	2986

951907	109956	109971	ACCAATAACTGACCTC	54	N/A	N/A	2987

951908	111211	111226	AGTTATTCTGATGCCA	3	N/A	N/A	2988

951909	111349	111364	GGGTATGGTTGCAACC	92	N/A	N/A	2989

951910	111384	111399	TCATTAGGACACAGCG	20	N/A	N/A	2990

951911	111385	111400	ATCATTAGGACACAGC	6	N/A	N/A	2991

951912	111494	111509	TACTATCAGTACAGCT	83	N/A	N/A	2992

951913	111496	111511	GATACTATCAGTACAG	42	N/A	N/A	2993

951914	111499	111514	GGAGATACTATCAGTA	115	N/A	N/A	2994

951915	111500	111515	TGGAGATACTATCAGT	27	N/A	N/A	2995

951916	112149	112164	GTGAACACTGATCACT	86	N/A	N/A	2996

951917	112185	112200	GGTATATGAGGAAACA	18	N/A	N/A	2997

951918	112208	112223	CCGTATATGTCTCCGA	8	N/A	N/A	2998

951919	112209	112224	ACCGTATATGTCTCCG	7	N/A	N/A	2999

951920	112293	112308	GGCAATTGTCTCTCTT	47	N/A	N/A	3000

951921	112316	112331	TGTATAAGCTCACTTT	40	N/A	N/A	3001

951922	112318	112333	CATGTATAAGCTCACT	105	N/A	N/A	3002

951923	112320	112335	AGCATGTATAAGCTCA	36	N/A	N/A	3003

951924	112369	112384	ATAATTATGTCCCATC	24	N/A	N/A	3004

951925	112370	112385	GATAATTATGTCCCAT	5	N/A	N/A	3005

951926	112371	112386	GGATAATTATGTCCCA	61	N/A	N/A	3006

951927	112372	112387	TGGATAATTATGTCCC	102	N/A	N/A	3007

951928	112373	112388	ATGGATAATTATGTCC	105	N/A	N/A	3008

951929	112950	112965	ACTACTAGTTTCTGTT	91	N/A	N/A	3009

951930	112951	112966	CACTACTAGTTTCTGT	37	N/A	N/A	3010

951931	113712	113727	GGTTATGAGATTCTTA	4	N/A	N/A	3011

951932	113713	113728	TGGTTATGAGATTCTT	5	N/A	N/A	3012

951933	114474	114489	TTTATTCACAGGGTCC	29	N/A	N/A	3013

951934	114476	114491	TATTTATTCACAGGGT	28	N/A	N/A	3014

951935	114477	114492	GTATTTATTCACAGGG	18	N/A	N/A	3015

951936	114510	114525	GCCAATGGAAGGCACC	129	N/A	N/A	3016

951937	114524	114539	TCCATTTGGGCTGTGC	86	N/A	N/A	3017

951938	114750	114765	TGCGATATCCTCCCCT	15	N/A	N/A	3018

951939	115717	115732	GAGAATAAAGCCTGGC	80	N/A	N/A	3019

908415	83669	83684	GATTGAGGTAAGGGTT	20	N/A	N/A	3020
	83757	83772			N/A	N/A
	83845	83860			N/A	N/A

908426	83689	83704	ACGGAACCGCTCTGTA	88	N/A	N/A	509
	83777	83792			N/A	N/A
	83865	83880			N/A	N/A

908428	83691	83706	TGACGGAACCGCTCTG	73	N/A	N/A	511
	83779	83794			N/A	N/A
	83867	83882			N/A	N/A

908507	103213	103228	AGCAGAGCATAGGAGA	20	N/A	N/A	590
	103251	103266			N/A	N/A

949039	7946	7961	AAGTGCTATTATAACT	100	N/A	N/A	3021
	8587	8602			N/A	N/A

949040	7947	7962	GAAGTGCTATTATAAC	8	N/A	N/A	3022
	8588	8603			N/A	N/A

949041	7948	7963	AGAAGTGCTATTATAA	6	N/A	N/A	3023
	8589	8604			N/A	N/A

949042	7949	7964	AAGAAGTGCTATTATA	15	N/A	N/A	3024
	8590	8605			N/A	N/A

949043	7950	7965	AAAGAAGTGCTATTAT	37	N/A	N/A	3025
	8591	8606			N/A	N/A

949044	7951	7966	GAAAGAAGTGCTATTA	19	N/A	N/A	3026
	8592	8607			N/A	N/A

949045	7952	7967	GGAAAGAAGTGCTATT	20	N/A	N/A	3027
	8593	8608			N/A	N/A

949046	7953	7968	GGGAAAGAAGTGCTAT	36	N/A	N/A	3028
	8594	8609			N/A	N/A

949047	7954	7969	AGGGAAAGAAGTGCTA	64	N/A	N/A	3029
	8595	8610			N/A	N/A

949048	7955	7970	AAGGGAAAGAAGTGCT	22	N/A	N/A	3030
	8596	8611			N/A	N/A

949049	7956	7971	AAAGGGAAAGAAGTGC	21	N/A	N/A	3031
	8597	8612			N/A	N/A

949060	9158	9173	TAAAATGTACCCCCTC	42	N/A	N/A	3032
	9478	9493			N/A	N/A

949133	16330	16345	GAAAAGAACAACCTGG	21	N/A	N/A	3033
	16655	16670			N/A	N/A

949913	83642	83657	TGCAGCTGTCAGCATG	61	N/A	N/A	3034
	83730	83745			N/A	N/A
	83818	83833			N/A	N/A

949914	83643	83658	ATGCAGCTGTCAGCAT	113	N/A	N/A	3035
	83731	83746			N/A	N/A
	83819	83834			N/A	N/A

949915	83644	83659	TATGCAGCTGTCAGCA	59	N/A	N/A	3036
	83732	83747			N/A	N/A
	83820	83835			N/A	N/A

949916	83645	83660	ATATGCAGCTGTCAGC	16	N/A	N/A	3037
	83733	83748			N/A	N/A
	83821	83836			N/A	N/A

949917	83648	83663	GCCATATGCAGCTGTC	13	N/A	N/A	3038
	83736	83751			N/A	N/A
	83824	83839			N/A	N/A

949918	83652	83667	CTTTGCCATATGCAGC	8	N/A	N/A	3039
	83740	83755			N/A	N/A
	83828	83843			N/A	N/A

949919	83653	83668	ACTTTGCCATATGCAG	15	N/A	N/A	3040
	83741	83756			N/A	N/A
	83829	83844			N/A	N/A

949920	83654	83669	TACTTTGCCATATGCA	4	N/A	N/A	3041
	83742	83757			N/A	N/A
	83830	83845			N/A	N/A

949921	83655	83670	TTACTTTGCCATATGC	1	N/A	N/A	3042
	83743	83758			N/A	N/A
	83831	83846			N/A	N/A

949922	83656	83671	GTTACTTTGCCATATG	4	N/A	N/A	3043
	83744	83759			N/A	N/A
	83832	83847			N/A	N/A

949923	83657	83672	GGTTACTTTGCCATAT	8	N/A	N/A	3044
	83745	83760			N/A	N/A
	83833	83848			N/A	N/A

949924	83668	83683	ATTGAGGTAAGGGTTA	4	N/A	N/A	3045
	83756	83771			N/A	N/A
	83844	83859			N/A	N/A

949925	83675	83690	TATCTTGATTGAGGTA	6	N/A	N/A	3046
	83763	83778			N/A	N/A
	83851	83866			N/A	N/A

949926	83676	83691	GTATCTTGATTGAGGT	2	N/A	N/A	3047
	83764	83779			N/A	N/A
	83852	83867			N/A	N/A

949927	83677	83692	TGTATCTTGATTGAGG	1	N/A	N/A	3048
	83765	83780			N/A	N/A
	83853	83868			N/A	N/A

949928	83678	83693	CTGTATCTTGATTGAG	4	N/A	N/A	3049
	83766	83781			N/A	N/A
	83854	83869			N/A	N/A

949929	83682	83697	CGCTCTGTATCTTGAT	2	N/A	N/A	3050
	83770	83785			N/A	N/A
	83858	83873			N/A	N/A

949930	83683	83698	CCGCTCTGTATCTTGA	9	N/A	N/A	3051
	83771	83786			N/A	N/A
	83859	83874			N/A	N/A

949931	83700	83715	TTTAGGAGGTGACGGA	6	N/A	N/A	3052
	83788	83803			N/A	N/A
	83876	83891			N/A	N/A

949932	83701	83716	TTTTAGGAGGTGACGG	7	N/A	N/A	3053
	83789	83804			N/A	N/A
	83877	83892			N/A	N/A

949933	83702	83717	ATTTTAGGAGGTGACG	17	N/A	N/A	3054
	83878	83893			N/A	N/A

949934	83703	83718	GATTTTAGGAGGTGAC	15	N/A	N/A	3055
	83879	83894			N/A	N/A

949935	83704	83719	GGATTTTAGGAGGTGA	6	N/A	N/A	3056
	83880	83895			N/A	N/A

949936	83705	83720	GGGATTTTAGGAGGTG	15	N/A	N/A	3057
	83881	83896			N/A	N/A

949937	83706	83721	AGGGATTTTAGGAGGT	8	N/A	N/A	3058
	83882	83897			N/A	N/A

949938	83707	83722	CAGGGATTTTAGGAGG	12	N/A	N/A	3059
	83883	83898			N/A	N/A

949939	83708	83723	ACAGGGATTTTAGGAG	18	N/A	N/A	3060
	83884	83899			N/A	N/A

949940	83728	83743	CAGCTGTCAGCATGAT	63	N/A	N/A	3061
	83816	83831			N/A	N/A

949941	83729	83744	GCAGCTGTCAGCATGA	67	N/A	N/A	3062
	83817	83832			N/A	N/A

950194	103191	103206	CGGTGGCCATGGAGGG	67	N/A	N/A	3063
	103229	103244			N/A	N/A
	103267	103282			N/A	N/A

950195	103192	103207	CCGGTGGCCATGGAGG	98	N/A	N/A	3064
	103230	103245			N/A	N/A
	103268	103283			N/A	N/A

950196	103193	103208	CCCGGTGGCCATGGAG	91	N/A	N/A	3065
	103231	103246			N/A	N/A
	103269	103284			N/A	N/A

950197	103194	103209	GCCCGGTGGCCATGGA	122	N/A	N/A	3066
	103232	103247			N/A	N/A
	103270	103285			N/A	N/A

950198	103195	103210	GGCCCGGTGGCCATGG	106	N/A	N/A	3067
	103233	103248			N/A	N/A
	103271	103286			N/A	N/A

950199	103196	103211	GGGCCCGGTGGCCATG	125	N/A	N/A	3068
	103234	103249			N/A	N/A
	103272	103287			N/A	N/A

950200	103210	103225	AGAGCATAGGAGAGGG	20	N/A	N/A	3069
	103248	103263			N/A	N/A

950201	103212	103227	GCAGAGCATAGGAGAG	21	N/A	N/A	3070
	103250	103265			N/A	N/A

950202	103214	103229	GAGCAGAGCATAGGAG	23	N/A	N/A	3071
	103252	103267			N/A	N/A

950203	103215	103230	GGAGCAGAGCATAGGA	22	N/A	N/A	3072
	103253	103268			N/A	N/A

950204	103217	103232	AGGGAGCAGAGCATAG	53	N/A	N/A	3073
	103255	103270			N/A	N/A

950205	103219	103234	GGAGGGAGCAGAGCAT	50	N/A	N/A	3074
	103257	103272			N/A	N/A

950206	103221	103236	ATGGAGGGAGCAGAGC	53	N/A	N/A	3075
	103259	103274			N/A	N/A

950297	113289	113304	ATCCTGTTATATCTTT	3	N/A	N/A	3076
	113606	113621			N/A	N/A

950298	113290	113305	AATCCTGTTATATCTT	2	N/A	N/A	3077
	113607	113622			N/A	N/A

Each modified oligonucleotide listed in the table below is 100% complementary to human DNM2 nucleic acid sequence GENBANK Number NM_001005361.2. (SEQ ID No: 3). As shown below, modified oligonucleotides complementary to human DNM2 inhibited human DNM2 mRNA expression.

TABLE 4

DNM2 mRNA Expression

	SEQ	SEQ
	ID: 3	ID: 3	Sequence	Dynamin2	SEQ
Compound	Start	Stop		(%	ID
Number	Site	Site		Control)	NO

948482	1735	1750	CACCAGGATCTCCCCC	30	3078

948483	1738	1753	GATCACCAGGATCTCC	101	3079

948509	1511	1526	GCTTCTCGGCACACTT	27	3080

948510	1512	1527	AGCTTCTCGGCACACT	48	3081

948511	1514	1529	TGAGCTTCTCGGCACA	111	3082

948512	1520	1535	AGGAACTGAGCTTCTC	39	3083

948513	1521	1536	TAGGAACTGAGCTTCT	37	3084

Example 3: Effect of Modified Oligonucleotides Complementary to Dynamin 2 In Vitro

Modified oligonucleotides complementary to dynamin 2 (DNM2) nucleic acid were designed and tested for their effect on dynamin 2 mRNA in vitro. The modified oligonucleotides were tested in a series of experiments that had similar culture conditions.
A431 cells cultured at a density of 10,000 cells per well were treated with only 500 nM of modified oligonucleotide via free uptake or no modified oligonucleotide for untreated controls. After approximately 24 hours, RNA was isolated from the cells and DNM2 mRNA levels were measured by quantitative real-time PCR as described in Example 1.
The modified oligonucleotides described in the tables below each have a 3-10-3 cEt gapmer motif, wherein the central gap segment contains ten 2′-deoxynucleosides and is flanked by wing segments on the 3′ and 5′ ends, each containing three cEt nucleosides. Each modified oligonucleotide listed in the tables below is 100% complementary to human DNM2 nucleic acid sequence GENBANK Number NC_000019.10, truncated from 10715001 to 1083500 (SEQ ID NO:1) and/or GENBANK Number NM_004945.3 (SEQ ID NO: 2). As shown below, modified oligonucleotides complementary to human DNM2 inhibited human DNM2 mRNA expression.

TABLE 5

DNM2 mRNA Expression

					SEQ	SEQ
	SEQ ID:	SEQ ID:			ID: 2	ID 2:	SEQ
Compound	1 Start	1 Stop		DNM2	Start	Stop	ID
Number	Site	Site	Sequence	(% Control)	Site	Site	NO

694824	57496	57511	TTTTGGACTTGCAGTG	25	443	458	3085

695171	82451	82466	ACACACTTCAAACTCG	30	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	26	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	28	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	31	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	27	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	42	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	37	N/A	N/A	13

695171	82451	82466	ACACACTTCAAACTCG	29	N/A	N/A	13

951371	11922	11937	GTTGATAGTCGGTTGT	9	N/A	N/A	2452

951371	11922	11937	GTTGATAGTCGGTTGT	8	N/A	N/A	2452

951371	11922	11937	GTTGATAGTCGGTTGT	7	N/A	N/A	2452

951371	11922	11937	GTTGATAGTCGGTTGT	7	N/A	N/A	2452

951371	11922	11937	GTTGATAGTCGGTTGT	7	N/A	N/A	2452

951371	11922	11937	GTTGATAGTCGGTTGT	10	N/A	N/A	2452

951371	11922	11937	GTTGATAGTCGGTTGT	19	N/A	N/A	2452

951371	11922	11937	GTTGATAGTCGGTTGT	9	N/A	N/A	2452

988378	82447	82462	ACTTCAAACTCGGCTC	24	N/A	N/A	3086

988379	81166	81181	AACTGTATTGATTAGC	20	1492	1507	3087

988380	116569	116584	GTGTGGTTAATATGGC	2	3313	3328	3088

988381	116853	116868	ATAGAATACAGAGTGC	19	3597	3612	3089

988382	11917	11932	TAGTCGGTTGTCTGGA	21	N/A	N/A	3090

988383	11919	11934	GATAGTCGGTTGTCTG	37	N/A	N/A	3091

988384	11920	11935	TGATAGTCGGTTGTCT	67	N/A	N/A	3092

988385	11921	11936	TTGATAGTCGGTTGTC	26	N/A	N/A	3093

988386	11923	11938	AGTTGATAGTCGGTTG	3	N/A	N/A	3094

988387	11924	11939	GAGTTGATAGTCGGTT	5	N/A	N/A	3095

988388	11925	11940	CGAGTTGATAGTCGGT	9	N/A	N/A	3096

988389	11927	11942	AGCGAGTTGATAGTCG	42	N/A	N/A	3097

988390	24283	24298	GTATTATGGTATGTGA	14	N/A	N/A	3098

988391	24286	24301	TAGGTATTATGGTATG	20	N/A	N/A	3099

988392	24287	24302	TTAGGTATTATGGTAT	31	N/A	N/A	3100

988393	24288	24303	ATTAGGTATTATGGTA	29	N/A	N/A	3101

988394	24290	24305	GGATTAGGTATTATGG	6	N/A	N/A	3102

988395	82040	82055	TTGGACTGTTTTCCCC	39	N/A	N/A	3103

988396	82042	82057	GTTTGGACTGTTTTCC	7	N/A	N/A	3104

988397	82824	82839	GATGTGTTTCCTGTGT	2	N/A	N/A	3105

988398	82825	82840	TGATGTGTTTCCTGTG	3	N/A	N/A	3106

988399	82826	82841	ATGATGTGTTTCCTGT	5	N/A	N/A	3107

988400	82828	82843	AGATGATGTGTTTCCT	4	N/A	N/A	3108

988401	82829	82844	CAGATGATGTGTTTCC	4	N/A	N/A	3109

988402	82830	82845	ACAGATGATGTGTTTC	7	N/A	N/A	3110

988403	82832	82847	ATACAGATGATGTGTT	99	N/A	N/A	3111

988404	83917	83932	TTTTTGAGAGGTTGAC	19	N/A	N/A	3112

988405	89716	89731	TATAGGGCTTCTTATC	61	N/A	N/A	3113

988406	89717	89732	TTATAGGGCTTCTTAT	64	N/A	N/A	3114

988407	89723	89738	AGTTTATTATAGGGCT	8	N/A	N/A	3115

988408	89724	89739	AAGTTTATTATAGGGC	13	N/A	N/A	3116

988409	89725	89740	TAAGTTTATTATAGGG	25	N/A	N/A	3117

988410	89726	89741	GTAAGTTTATTATAGG	24	N/A	N/A	3118

988411	89727	89742	AGTAAGTTTATTATAG	96	N/A	N/A	3119

988412	90717	90732	ATGTATCTGGTTCAAT	34	N/A	N/A	3120

988413	90718	90733	AATGTATCTGGTTCAA	11	N/A	N/A	3121

988414	90719	90734	AAATGTATCTGGTTCA	8	N/A	N/A	3122

988415	90720	90735	TAAATGTATCTGGTTC	11	N/A	N/A	3123

988416	90723	90738	AAGTAAATGTATCTGG	15	N/A	N/A	3124

988417	90724	90739	CAAGTAAATGTATCTG	51	N/A	N/A	3125

988418	90725	90740	ACAAGTAAATGTATCT	45	N/A	N/A	3126

988419	90726	90741	AACAAGTAAATGTATC	109	N/A	N/A	3127

988420	93918	93933	TTCTGGTCTGGCTTAA	16	N/A	N/A	3128

988421	93920	93935	AATTCTGGTCTGGCTT	45	N/A	N/A	3129

988422	93922	93937	TGAATTCTGGTCTGGC	7	N/A	N/A	3130

988423	93924	93939	AATGAATTCTGGTCTG	30	N/A	N/A	3131

988424	93925	93940	GAATGAATTCTGGTCT	80	N/A	N/A	3132

988425	93926	93941	AGAATGAATTCTGGTC	20	N/A	N/A	3133

988426	93928	93943	CAAGAATGAATTCTGG	74	N/A	N/A	3134

Example 4: Dose Response of Modified Oligonucleotides Complementary to Dynamin 2 In Vitro

Selected oligonucleotides listed in the Examples above were tested at various doses via free uptake in A431 cells. Cells were cultured at a density of 10,000 cells per well and treated with 62.5, 250, 1,000, or 4,000 nM of modified oligonucleotide, as specified in the table below. After a treatment period of approximately 48 hours, total RNA was isolated and analyzed by RT-PCR with primer probe set RTS36027 (described in Example 1), normalized with ribogreen. As illustrated in the tables below, DNM2 mRNA expression was inhibited in a dose-dependent manner by modified oligonucleotides complementary to DNM2.

TABLE 6

DNM2 mRNA Expression

Compound

DNM2 expression (% control)

Number	62.5 nM	250 nM	1,000 nM	4,000 nM	IC₅₀(μM)

694838	31	12	5	3	<0.1
695171	74	28	11	4	0.1
695171	83	33	11	5	0.2
695171	57	25	7	3	0.1
695171	59	24	9	4	0.1
695171	51	17	9	4	<0.1
695171	57	28	10	4	0.1
907945	38	10	5	3	<0.1
907946	70	36	17	11	0.2
907947	48	28	9	6	<0.1
907948	51	22	9	5	<0.1
907949	77	43	16	6	0.2
907966	53	30	15	8	<0.1
907967	32	10	6	4	<0.1
907968	65	30	14	8	0.1
907969	19	6	3	2	<0.1
907970	20	5	3	2	<0.1
907971	59	30	17	12	0.1
907972	72	36	18	17	0.2
907975	35	15	7	4	<0.1
907979	55	26	14	10	<0.1
907983	39	16	9	8	<0.1
907998	40	13	6	4	<0.1
908000	71	42	19	11	0.2
908011	53	24	13	9	<0.1
908013	52	25	13	7	<0.1
908025	69	31	13	6	0.1
908027	38	13	5	2	<0.1
908028	80	51	22	9	0.3
908029	83	43	19	9	0.3
908031	44	13	4	2	<0.1
908033	52	18	8	5	<0.1
908034	63	29	13	8	0.1
908035	26	6	2	1	<0.1
908036	71	39	20	11	0.2
908037	26	9	3	1	<0.1
908041	49	19	6	3	<0.1
908043	28	15	10	8	<0.1
908044	43	17	7	3	<0.1
908047	54	28	15	9	<0.1
908049	49	15	6	3	<0.1
908052	53	21	7	4	<0.1
908055	69	36	19	12	0.2
908056	54	23	9	6	<0.1
908059	35	11	3	2	<0.1
908061	27	7	2	1	<0.1
908068	72	41	21	10	0.2
908070	64	35	15	9	0.1
908085	68	33	18	11	0.1
908098	61	28	12	10	0.1
908102	42	28	17	11	<0.1
908104	44	23	12	8	<0.1
908107	51	25	11	7	<0.1
908135	46	23	14	14	<0.1
908138	50	24	11	6	<0.1
908146	58	33	19	15	0.1
908147	37	18	10	6	<0.1
908148	42	15	7	5	<0.1
908154	28	9	3	2	<0.1
908164	65	31	16	10	0.1
908166	44	18	9	5	<0.1
908167	62	28	16	12	0.1
908177	69	37	15	7	0.2
908178	52	22	6	4	<0.1
908179	47	18	7	4	<0.1
908183	58	34	15	11	0.1
908184	56	24	11	8	<0.1
908201	26	13	5	4	<0.1
908213	37	13	5	3	<0.1
908214	57	37	17	12	0.1
908215	63	30	12	6	0.1
908217	32	10	3	2	<0.1
908232	58	26	6	3	0.1
908255	64	28	13	9	0.1
908267	48	28	12	8	<0.1
908355	83	62	42	26	0.6
908389	43	16	5	3	<0.1
908392	12	3	1	1	<0.1
908396	57	19	6	3	<0.1
908401	26	6	2	1	<0.1
908404	58	29	15	8	0.1
908405	49	26	9	4	<0.1
908407	70	30	12	5	0.1
908408	18	2	1	1	<0.1
908409	67	33	13	5	0.1
908410	71	36	15	7	0.2
908412	58	24	10	6	0.1
908413	33	8	2	1	<0.1
908414	25	6	3	2	<0.1
908415	40	11	4	3	<0.1
908416	42	13	4	1	<0.1
908417	26	5	2	1	<0.1
908418	63	33	11	4	0.1
908419	50	11	4	2	<0.1
908420	17	3	1	1	<0.1
908421	23	4	1	1	<0.1
908422	51	16	5	3	<0.1
908424	57	22	9	7	<0.1
908425	57	38	20	12	0.1
908434	54	20	8	3	<0.1
908436	71	40	17	8	0.2
908439	38	9	2	2	<0.1
908440	25	6	3	3	<0.1
908441	33	6	2	1	<0.1
908442	32	9	3	1	<0.1
908443	56	17	5	2	<0.1
908444	31	7	2	1	<0.1
908448	78	44	13	8	0.2
908449	47	20	7	4	<0.1
908451	45	14	6	5	<0.1
908475	66	29	12	5	0.1
908506	34	9	4	3	<0.1
908507	67	39	16	10	0.1
908508	52	30	14	10	<0.1

Example 5: Dose Response of Modified Oligonucleotides Complementary to Dynamin 2 In Vitro

Selected oligonucleotides listed in the Examples above were tested via free uptake at various doses in A431 cells. Cells were cultured at a density of 10,000 cells per well and treated with 31.25, 125, 500, or 2,000 nM of modified oligonucleotide, as specified in the tables below. After approximately 48 hours, total RNA was isolated and analyzed by RT-PCR with primer probe set RTS36027 (described in Example 1), normalized with ribogreen. As illustrated in the tables below, DNM2 mRNA expression was inhibited in a dose-dependent manner by modified oligonucleotides complementary to DNM2.

TABLE 7

DNM2 mRNA Expression

Compound

DNM2 expression (% control)

Number	31.25 nM	125 nM	500 nM	2,000 nM	IC50 (μM)

695015	105	94	89	94	>2.0
695170	62	30	13	5	0.05
695171	72	33	11	4	0.07
695171	70	33	10	3	0.1
695171	70	35	14	6	0.1
695171	58	22	9	2	<0.03
695171	71	36	12	4	0.08
695171	70	32	11	4	0.07
695171	74	36	12	4	0.08
695171	71	31	10	3	0.07
695171	73	36	14	4	0.08
695171	73	36	17	7	0.09
695171	54	27	10	3	<0.03
695171	69	33	10	5	0.06
695171	74	35	12	4	0.08
695171	81	40	14	4	0.11
695171	79	40	14	5	0.11
695171	77	38	14	5	0.10
695171	72	37	13	5	0.08
695171	71	34	11	4	0.07
695171	66	43	11	5	0.08
695171	68	33	11	4	0.06
695171	63	29	12	5	0.05
948488	79	33	12	4	0.09
948489	53	17	5	1	<0.03
948490	25	5	1	0	<0.03
948492	67	32	9	4	0.06
948496	72	43	15	6	0.09
948505	67	36	12	5	0.07
948506	58	16	5	2	<0.03
948507	80	30	9	3	0.1
948508	63	26	7	3	0.0
948521	79	35	19	13	0.11
948524	70	43	19	9	0.09
948525	75	47	19	7	0.12
948526	73	35	12	7	0.08
948527	62	31	14	9	0.05
948528	64	29	11	20	0.04
948538	104	95	96	91	>2.0
948549	71	38	16	11	0.08
948556	36	11	4	3	<0.03
948557	23	8	4	3	<0.03
948558	68	29	14	10	0.06
948559	48	16	7	5	<0.03
948560	24	7	3	2	<0.03
948562	69	26	10	5	0.06
948568	57	26	11	7	<0.03
948575	73	39	20	10	0.1
948587	43	14	7	4	<0.03
948589	66	21	11	9	0.04
948590	53	35	16	10	0.03
948602	39	15	7	5	<0.03
948604	42	17	7	4	<0.03
948608	54	18	6	4	<0.03
948622	79	47	22	10	0.13
948629	51	13	6	2	<0.03
948630	67	33	13	5	0.06
948631	46	14	5	3	<0.03
948632	42	12	5	2	<0.03
948633	57	21	8	6	<0.03
948646	63	58	23	10	0.11
948654	68	40	13	4	0.08
948656	69	40	22	13	0.09
948660	47	11	2	1	<0.03
948661	80	41	14	4	0.11
948662	63	38	16	8	0.06
948664	45	12	2	1	<0.03
948665	64	25	7	3	0.04
948666	82	37	12	4	0.11
948673	33	9	3	2	<0.03
948674	75	39	12	5	0.1
948675	43	8	2	0	<0.03
948676	40	10	2	0	<0.03
948677	53	11	2	1	<0.03
948678	69	27	9	4	0.06
948684	61	29	11	4	0.04
948685	52	15	5	2	<0.03
948686	24	8	2	1	<0.03
948687	58	19	5	2	<0.03
948689	73	26	17	8	0.07
948692	49	20	8	6	<0.03
948693	71	40	18	10	0.09
948695	75	38	20	7	0.1
948700	63	25	7	3	0.04
948701	64	24	9	4	0.04
948702	60	25	6	1	0.04
948703	66	29	7	2	0.05
948707	69	24	7	3	0.05
948729	47	12	2	1	<0.03
948731	67	33	11	4	0.06
948732	68	40	14	7	0.08
948735	22	9	2	2	<0.03
948741	49	55	24	15	0.06
948752	64	31	12	5	0.05
948765	61	20	7	3	0.03
948766	78	43	18	7	0.11
948767	76	40	19	12	0.1
948768	67	35	17	12	0.1
948769	60	24	10	5	0.0
948788	57	27	13	10	0.03
948794	51	26	5	2	<0.03
948804	42	18	8	5	<0.03
948805	34	13	5	3	<0.03
948806	16	18	7	5	<0.03
948807	56	35	18	11	0.04
948812	77	40	20	11	0.11
948835	66	28	7	3	0.05
948836	52	27	11	6	<0.03
948847	68	32	12	7	0.06
948849	43	13	6	5	<0.03
948850	43	15	7	5	<0.03
948851	45	16	5	3	<0.03
948852	51	16	4	2	<0.03
948853	84	32	9	5	0.10
948854	58	21	7	3	<0.03
948855	56	30	12	6	0.03
948856	70	38	14	8	0.08
948862	55	26	10	4	<0.03
948863	43	8	2	2	<0.03
948865	57	24	10	6	<0.03
948880	84	46	26	14	0.16
948882	66	36	19	8	0.07
948883	81	62	29	16	0.21
948884	59	25	9	4	0.03
948885	53	18	6	4	<0.03
948891	96	52	24	12	0.20
948892	54	24	7	4	<0.03
948893	93	84	59	44	1.24
948906	81	46	19	9	0.13
948907	65	25	8	4	0.05
948910	62	22	8	4	0.0
948911	48	16	10	4	<0.03
948917	81	57	30	19	0.20
948918	69	36	14	6	0.07
948922	64	32	12	6	0.05
948923	30	8	4	3	<0.03
948924	32	11	3	2	<0.03
948925	58	27	9	6	0.03
948926	109	94	105	111	>2.0
948935	38	17	6	3	<0.03
948936	47	12	4	2	<0.03
948937	48	21	7	3	<0.03
948938	49	19	8	5	<0.03
948950	54	27	8	3	<0.03
948951	32	13	5	3	<0.03
948952	46	13	4	2	<0.03
948953	69	41	14	7	0.08
948954	43	15	8	4	<0.03
948960	41	14	8	7	<0.03
948961	48	17	6	3	<0.03
948963	51	23	8	4	<0.03
948965	96	77	53	28	0.57
948966	63	32	11	4	0.05
948967	71	38	13	6	0.08
948968	60	29	9	3	0.04
948969	47	18	6	4	<0.03
948970	31	9	3	2	<0.03
948971	34	9	2	2	<0.03
948972	72	32	9	3	0.07
949006	79	53	19	8	0.1
949007	84	37	9	6	0.1
949012	81	38	17	10	0.11
949040	80	36	11	4	0.10
949041	101	50	14	5	0.18
949051	74	45	16	8	0.10
949053	89	54	18	5	0.2
949055	68	24	8	4	0.0
949059	73	40	15	5	0.09
949061	109	99	83	73	>2.0
949077	82	47	15	5	0.1
949080	48	20	7	5	<0.03
949088	73	24	8	3	0.06
949092	66	24	13	14	0.05
949094	79	48	17	6	0.13
949095	90	66	19	6	0.20
949109	95	55	26	13	0.22
949139	72	47	24	11	0.12
949158	54	20	10	8	<0.03
949159	75	40	15	25	0.10
949163	52	15	4	2	<0.03
949164	42	30	15	10	<0.03
949167	78	39	17	21	0.11
949168	67	36	19	12	0.07
949182	65	27	8	4	0.05
949190	57	15	3	2	<0.03
949216	74	42	14	5	0.10
949223	87	28	16	6	0.1
949229	125	101	92	78	>2.0
949279	85	41	15	5	0.12
949325	98	112	86	75	>2.0
949397	94	96	100	106	>2.0
949483	71	43	21	11	0.10
949500	78	56	24	10	0.16
949517	103	98	97	80	>2.0
949589	111	89	57	42	1.06
949685	101	82	58	40	0.99
949778	90	55	20	9	0.18
949805	106	88	66	48	1.73
949814	59	51	22	8	0.08
949841	87	46	11	4	0.13
949846	48	30	11	7	<0.03
949848	57	18	4	2	<0.03
949858	66	23	10	6	0.04
949865	61	11	3	1	<0.03
949871	70	26	7	3	0.1
949872	47	13	3	1	<0.03
949873	45	10	2	1	<0.03
949874	14	3	1	0	<0.03
949875	54	23	5	2	<0.03
949876	79	32	10	5	0.09
949880	52	20	6	3	<0.03
949884	87	41	16	7	0.13
949886	62	22	6	3	0.04
949887	87	64	21	8	0.19
949892	83	49	22	13	0.2
949896	13	1	1	0	<0.03
949897	67	32	15	7	0.06
949899	80	49	20	6	0.14
949917	80	48	22	10	0.1
949918	84	46	15	4	0.1
949920	59	20	3	2	<0.03
949921	67	19	2	1	0.04
949922	56	21	6	2	<0.03
949923	70	34	15	7	0.07
949924	112	106	5	2	0.31
949925	59	27	6	2	0.04
949926	44	7	1	1	<0.03
949927	37	4	1	1	<0.03
949928	68	20	4	2	0.04
949929	66	18	3	1	0.04
949930	61	30	10	4	0.04
949931	68	27	6	2	0.05
949932	72	31	6	2	0.07
949934	66	42	22	12	0.08
949935	52	20	8	6	<0.03
949937	69	27	7	5	0.06
949938	66	35	14	5	0.06
949942	69	30	8	3	0.06
949943	52	21	6	2	<0.03
949944	60	20	4	3	0.03
949945	54	16	5	3	<0.03
949948	34	5	1	1	<0.03
949949	46	10	2	1	<0.03
949950	46	12	3	2	<0.03
949962	70	34	16	9	0.1
949963	71	43	15	6	0.1
949978	65	35	9	2	0.06
949979	72	35	11	5	0.08
949983	89	66	25	12	0.23
949990	61	23	5	2	0.04
949991	55	21	5	1	<0.03
949992	56	29	11	6	0.03
950014	60	28	8	3	0.04
950017	68	38	11	5	0.07
950019	52	20	8	5	<0.03
950020	84	43	13	5	0.12
950021	80	46	15	6	0.12
950023	57	17	5	2	<0.03
950024	51	17	7	5	<0.03
950030	37	7	1	0	<0.03
950048	57	40	16	6	0.05
950057	65	38	21	9	0.07
950060	26	16	3	2	<0.03
950061	55	34	10	5	0.04
950064	74	32	11	4	0.08
950065	72	30	8	2	0.07
950069	68	31	8	4	0.06
950072	49	20	6	3	<0.03
950075	87	38	13	5	0.12
950080	69	40	15	7	0.08
950081	60	28	8	4	0.04
950083	65	29	12	4	0.1
950085	53	14	6	4	<0.03
950086	76	43	15	4	0.10
950089	46	10	2	1	<0.03
950091	45	12	2	1	<0.03
950092	80	42	14	8	0.11
950102	81	37	9	3	0.10
950124	69	31	14	8	0.06
950132	55	14	4	1	<0.03
950139	87	59	24	9	0.19
950183	44	23	5	2	<0.03
950219	84	37	18	11	0.12
950237	83	48	17	8	0.14
950246	76	33	15	7	0.09
951328	76	37	12	4	0.09
951330	76	40	13	5	0.10
951333	89	45	22	9	0.16
951340	74	42	15	6	0.10
951341	92	58	21	4	0.19
951346	49	22	4	3	<0.03
951352	67	19	7	3	0.04
951354	67	36	12	6	0.07
951356	57	23	6	2	<0.03
951357	62	35	10	4	0.05
951358	76	36	12	5	0.09
951362	53	15	3	1	<0.03
951364	58	18	4	2	<0.03
951365	78	47	16	6	0.12
951366	61	34	12	6	0.05
951367	69	28	8	4	0.06
951370	74	41	16	7	0.10
951371	30	7	2	2	<0.03
951372	61	21	6	4	0.03
951374	62	17	4	2	0.03
951376	67	47	15	5	0.09
951381	63	28	11	6	0.05
951384	63	27	6	2	0.05
951386	66	22	6	3	0.04
951387	68	20	3	1	0.04
951397	69	35	8	4	0.07
951406	73	52	19	9	0.12
951407	56	21	6	3	<0.03
951408	47	15	3	1	<0.03
951413	68	46	20	9	0.09
951416	63	42	17	7	0.07
951418	60	22	6	3	0.03
951428	28	4	1	1	<0.03
951429	32	6	2	2	<0.03
951430	59	25	6	2	0.03
951431	46	32	9	5	<0.03
951441	100	82	67	42	1.33
951450	71	36	12	6	0.08
951458	63	32	15	10	0.05
951459	77	44	17	9	0.11
951462	74	28	6	2	0.07
951463	74	35	15	6	0.09
951481	91	47	26	13	0.18
951489	59	17	8	3	<0.03
951494	64	32	10	5	0.05
951498	58	27	10	7	0.03
951519	60	32	8	3	0.05
951529	90	35	24	18	0.15
951537	103	101	105	168	>2.0
951550	66	42	15	6	0.08
951554	83	36	10	5	0.10
951561	88	59	74	100	>2.0
951565	65	38	13	4	0.07
951566	77	36	9	3	0.09
951608	73	25	8	4	0.06
951645	64	29	12	7	0.05
951657	107	91	64	62	>2.0
951691	58	28	9	3	0.04
951696	58	19	9	5	<0.03
951700	76	40	16	9	0.10
951709	52	25	9	6	<0.03
951729	73	74	88	89	>2.0
951731	70	34	10	4	0.07
951737	57	24	5	4	<0.03
951746	68	31	17	11	0.06
951749	71	27	4	2	0.06
951763	44	11	4	2	<0.03
951764	40	9	2	1	<0.03
951767	87	48	16	5	0.14
951772	62	30	10	7	0.05
951781	95	42	15	7	0.15
951783	56	31	12	5	0.04
951787	53	20	7	4	<0.03
951793	44	11	4	2	<0.03
951795	64	24	7	3	0.04
951797	40	15	6	4	<0.03
951798	31	5	1	1	<0.03
951799	27	5	1	1	<0.03
951803	67	29	9	4	0.06
951804	45	13	3	1	<0.03
951806	65	43	15	7	0.07
951807	59	30	10	4	0.04
951808	69	46	18	6	0.10
951809	47	13	3	1	<0.03
951811	57	22	7	3	<0.03
951814	60	25	10	7	0.04
951815	58	37	15	7	0.05
951816	46	10	2	1	<0.03
951821	41	14	4	2	<0.03
951822	86	44	15	7	0.13
951824	67	23	5	3	0.05
951827	51	17	5	2	<0.03
951828	63	23	9	4	0.04
951830	61	26	10	5	0.04
951833	59	39	9	4	0.05
951835	23	4	1	0	<0.03
951837	53	16	4	1	<0.03
951838	80	51	22	8	0.14
951839	56	25	8	2	<0.03
951840	76	37	12	4	0.09
951841	69	27	11	2	0.06
951843	63	29	8	4	0.05
951845	39	9	3	1	<0.03
951846	45	14	3	1	<0.03
951855	87	65	19	5	0.19
951856	61	18	4	2	<0.03
951857	57	27	7	2	0.03
951874	74	73	68	47	>2.0
951875	66	33	10	4	0.06
951881	49	22	3	2	<0.03
951893	53	26	10	5	<0.03
951898	46	18	5	2	<0.03
951902	61	34	13	7	0.05
951938	85	51	20	11	0.16

Example 6: Dose Response of Modified Oligonucleotides Complementary to Dynamin 2 In Vitro

Selected oligonucleotides listed in the Examples above were tested at various doses via free uptake in A431 cells. Cells were cultured at a density of 10,000 cells per well and treated with 8, 40, 200, or 1,000 nM of modified oligonucleotide, as specified in the table below. After approximately 48 hours, total RNA was isolated and analyzed by RT-PCR with primer probe set RTS36027 (described in Example 1), normalized with GADPH. As illustrated in the tables below, DNM2 mRNA expression was inhibited in a dose-dependent manner by modified oligonucleotides complementary to DNM2.

TABLE 8

DNM2 mRNA Expression

Compound

DNM2 expression (% control)

Number	8 nM	40 nM	200 nM	1,000 nM	IC50 (μM)

695171	90	84	54	26	0.24
951371	105	67	25	9	0.10
951371	101	70	27	11	0.10
951371	109	63	16	4	0.08
951371	91	61	23	8	0.07
951371	94	78	25	9	0.10
951371	91	63	23	8	0.07
951371	59	66	23	8	0.04
951371	94	56	20	7	0.07
951371	88	67	26	10	0.08
988380	77	33	5	1	0.02
988386	93	70	21	6	0.08
988387	101	46	5	1	0.05
988388	97	80	39	11	0.13
988394	120	81	23	5	0.12
988396	113	87	30	9	0.14
988397	77	29	7	2	0.02
988398	86	46	11	3	0.04
988399	81	47	13	3	0.04
988400	88	54	15	3	0.05
988401	98	65	16	6	0.07
988402	94	48	11	2	0.05
988407	99	77	30	8	0.11
988413	84	58	23	7	0.06
988414	95	73	23	6	0.09
988415	82	62	25	7	0.06
988422	91	60	17	7	0.06

Example 7: Dose Response of Modified Oligonucleotides Complementary to Dynamin 2 In Vitro

Selected oligonucleotides listed in the Examples above were tested at various doses via free uptake in A431 cells. Cells were cultured at a density of 10,000 cells per well treated with 7.8, 31.25, 125, 500, or 2,000 nM of modified oligonucleotide, as specified in the tables below. After approximately 48 hours, total RNA was isolated and analyzed by RT-PCR with primer probe set RTS36027 (described in Example 1), normalized with GADPH. As illustrated in the tables below, DNM2 expression was inhibited in a dose-dependent manner by modified oligonucleotides complementary to DNM2.

TABLE 9

DNM2 mRNA Expression

DNM2 expression (% control)

	7.8 nM	31.25	125	500	2,000	IC50
Compound Number	nM	nM	nM	nM	nM	(μM)

908166	106	97	57	28	16	0.22
908213	139	85	56	30	16	0.21
908408	98	80	33	7	2	0.08
908420	91	86	45	13	4	0.11
948660	105	90	51	13	2	0.14
948677	101	101	61	23	8	0.21
948951	97	79	35	14	8	0.09
949190	107	101	54	22	8	0.18
949865	94	80	40	16	7	0.10
949935	88	84	58	31	19	0.20
950023	89	75	45	13	4	0.09
950060	81	79	48	29	11	0.13
950089	84	79	53	18	5	0.12
950132	87	83	44	19	9	0.11
950183	88	83	58	35	15	0.21

TABLE 10

DNM2 mRNA Expression

DNM2 expression (% control)

	7.8 nM	31.25	125	500	2,000	IC50
Compound Number	nM	nM	nM	nM	nM	(μM)

908408	98	80	34	10	2	0.08
951356	190	79	60	28	10	0.23
951362	97	85	63	30	11	0.21
951372	87	80	55	23	10	0.14
951431	100	90	68	38	21	0.31
951797	89	77	42	19	10	0.10
951798	89	58	23	6	1	0.04
951799	90	68	23	6	2	0.05
951816	100	77	38	10	3	0.09
951821	87	57	45	16	6	0.07
951827	94	79	45	17	5	0.11
951839	95	90	69	38	14	0.29
951845	92	71	31	8	2	0.06
951846	91	85	50	19	5	0.13
951856	97	86	54	22	7	0.15

Example 8: Dose Response of Modified Oligonucleotides Complementary to Dynamin 2 In Vitro

Selected oligonucleotides listed in the Examples above were tested at various doses in HSMM cells (human myoblast, Lonza CC-2580). Cells were cultured at a density of 20,000 cells per well and transfected via electroporation with 7.8, 31.25, 125, 500, or 2,000 nM of modified oligonucleotide, as specified in the tables below. After approximately 16 hours, total RNA was isolated and analyzed by RT-PCR with primer probe set RTS36027 (described in Example 1), normalized with ribogreen. As illustrated in the tables below, DNM2 mRNA expression was inhibited in a dose-dependent manner by modified oligonucleotides complementary to DNM2.

TABLE 11

DNM2 mRNA Expression

DNM2 expression (% control)

	7.8 nM	31.25	125	500	2,000	IC50
Compound Number	nM	nM	nM	nM	nM	(μM)

908166	104	95	98	66	42	1.29
908213	93	82	70	42	19	0.33
908408	111	100	74	38	17	0.36
908420	91	93	67	39	17	0.30
948660	88	86	83	42	15	0.40
948677	90	80	87	51	22	0.54
948951	88	74	57	31	12	0.16
949190	99	88	82	47	23	0.49
949865	132	106	92	39	19	0.50
949935	90	92	94	60	25	0.77
950023	113	97	86	40	15	0.43
950060	109	105	78	69	32	0.95
950089	111	102	80	54	21	0.58
950132	117	98	79	51	19	0.51
950183	95	110	84	59	23	0.70

TABLE 12

DNM2 mRNA Expression

DNM2 expression (% control)

	7.8 nM	31.25	125	500	2,000	IC50
Compound Number	nM	nM	nM	nM	nM	(μM)

908408	98	102	81	55	20	0.58
951356	95	102	94	74	41	1.45
951362	95	67	85	56	29	0.62
951372	95	91	86	60	27	0.75
951431	87	90	89	55	35	0.77
951797	94	94	98	72	41	1.40
951798	103	95	79	40	21	0.40
951799	104	100	81	47	20	0.50
951816	106	91	89	53	30	0.70
951821	122	106	66	32	22	0.32
951827	100	87	51	38	20	0.21
951839	123	94	99	72	31	1.17
951845	104	94	75	51	20	0.48
951846	112	91	80	59	27	0.69
951856	113	101	77	50	22	0.52

Example 9: Dose Response of Modified Oligonucleotides Complementary to Dynamin 2 In Vitro

Selected oligonucleotides listed in the Examples above were tested at various doses in SkMC cells (human skeletal muscle, Lonza CC-2561). Cells were cultured at a density of 20,000 cells per well and transfected via electroporation with 7.8, 31.25, 125, 500, or 2,000 nM of modified oligonucleotide, as specified in the tables below. After approximately 16 hours, total RNA was isolated and analyzed by RT-PCR with primer probe set RTS36027 (described in Example 1), normalized with ribogreen. As illustrated in the tables below, DNM2 mRNA expression was inhibited in a dose-dependent manner by modified oligonucleotides complementary to DNM2.

TABLE 13

DNM2 mRNA Expression

DNM2 expression (% control)

	7.8 nM	31.25	125	500	2,000	IC50
Compound Number	nM	nM	nM	nM	nM	(μM)

908166	90	84	87	43	13	0.41
908213	101	79	59	21	5	0.15
908408	94	72	57	21	3	0.13
908420	84	72	53	9	4	0.10
948660	95	90	64	12	2	0.16
948677	88	77	58	15	4	0.13
948951	83	67	41	12	5	0.07
949190	103	84	61	16	4	0.16
949865	130	106	55	10	4	0.17
949935	118	100	86	33	7	0.36
950023	129	98	57	11	3	0.17
950060	101	103	72	26	5	0.26
950089	112	78	65	20	4	0.17
950132	115	93	53	14	3	0.15
950183	114	92	70	24	5	0.23

TABLE 14

DNM2 mRNA Expression

DNM2 expression (% control)

	7.8 nM	31.25	125	500	2,000	IC50
Compound Number	nM	nM	nM	nM	nM	(μM)

908408	91	80	46	10	3	0.10
951356	103	98	90	52	15	0.55
951362	90	88	65	22	4	0.19
951372	99	88	65	16	4	0.18
951431	103	97	70	25	6	0.24
951797	102	94	69	25	7	0.23
951798	99	90	46	8	3	0.12
951799	101	90	49	12	3	0.13
951816	96	78	63	13	4	0.14
951821	104	79	47	12	3	0.11
951827	94	71	44	8	3	0.08
951839	114	103	76	32	6	0.32
951845	97	88	55	12	4	0.14
951846	90	86	41	18	4	0.11
951856	102	91	62	15	4	0.17

Example 10: Administration of Modified Oligonucleotides Complementary to Human Dynamin 2 to CD1 Mice

CD1® mice (Charles River, Mass.) were treated with modified oligonucleotides selected from Examples above and evaluated for changes in the levels of various plasma chemistry markers. Groups of 6 week old male CD1 mice were injected subcutaneously once a week for 6 weeks with 50 mg/kg of a modified oligonucleotide listed in the tables below (50 mg/kg/week dose). Each group contained 4 mice. One control group of male CD1 mice was injected subcutaneously once a week for 6 weeks with PBS. Mice were sacrificed 48 hours after the last dose, and organs and plasma were harvested for further analysis.
Levels of transaminases, albumin, BUN, and billirubin were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, N.Y.). Kidney, liver, and spleen weights were also measured. The results, presented in the tables below, show that many of the modified oligonucleotides complementary to DNM2 were well tolerated in vivo.

TABLE 15

Levels of plasma markers

ALT	AST	Albumin	BUN	Creatine	T. Bil.
(U/L)	(U/L)	(mg/dL)	(mg/d)	(mg/dL)	(g/dL)

PBS	48	82	3.36	27.2	0.12	0.66
549144	72	68	3.23	23.6	0.12	0.47
908166	259	108	3.16	20.4	0.11	0.38
908213	93	85	2.95	23.9	0.10	0.34
908389	1073	636	3.17	22.7	0.09	0.55
908408	67	70	2.97	26.2	0.11	0.32
908415	286	209	2.60	26.2	0.10	0.28
908416	425	336	2.35	26.4	0.07	0.19
908420	105	144	2.50	26.5	0.06	0.38
948660	112	165	2.59	21.8	0.08	0.26
948673	225	263	2.15	27.2	0.04	0.82
948675	100	132	2.25	21.4	0.06	0.22
948677	65	91	2.32	17.7	0.05	0.21
948951	107	126	2.46	21.4	0.07	0.22
949158	713	608	2.99	18.6	0.01	0.27
949190	64	62	2.52	22.1	0.07	0.20
949865	45	58	2.62	23.2	0.06	0.22
949873	89	163	2.33	20.7	0.04	0.21
949874	1079	853	2.77	20.7	0.05	0.46
949935	98	81	2.63	22.9	0.07	0.20
950023	42	57	2.52	22.2	0.04	0.20
950030	59	77	2.44	20.6	0.05	0.21

TABLE 16

Levels of plasma markers

Compound	ALT	AST	Albumin	BUN	Creatine	T. Bil.
No.	(U/L)	(U/L)	(mg/dL)	(mg/d)	(mg/dL)	(g/dL)

PBS	37	62	2.78	22.0	0.08	0.46
549144	64	79	2.64	24.0	0.08	0.29
950060	47	63	2.47	22.0	0.09	0.33
950089	40	56	2.65	21.0	0.10	0.22
950132	58	58	2.58	20.0	0.07	0.24
950183	49	62	2.38	21.0	0.06	0.29
951356	64	80	2.39	23.0	0.10	0.25
951362	60	63	2.74	22.0	0.09	0.18
951371	312	209	2.32	20.0	0.09	0.18
951372	59	99	2.54	25.0	0.07	0.21
951407	172	200	2.72	24.0	0.07	0.24
951431	64	80	2.48	23.0	0.08	0.22
951793	319	238	2.5	19.0	0.10	0.20
951797	41	63	2.6	24.0	0.11	0.26
951798	53	97	2.57	26.0	0.10	0.19
951799	44	97	2.44	25.0	0.08	0.19
951816	121	113	2.63	19.0	0.10	0.20
951821	79	80	2.39	23.0	0.07	0.17
951827	56	88	2.46	25.0	0.10	0.21
951839	45	50	2.52	23.0	0.08	0.20
951845	117	132	2.51	21.0	0.08	0.25
951846	35	47	2.59	21.0	0.10	0.21
951856	112	412	2.25	20.0	0.09	0.17

TABLE 17

Organ weights

	kidney	liver	spleen
	(g)	(g)	(g)

PBS	0.55	1.9	0.09
549144	0.52	1.9	0.10
908166	0.48	2.0	0.08
908213	0.52	2.0	0.12
908389	0.54	2.6	0.14
908408	0.49	2.2	0.13
908415	0.51	2.2	0.12
908416	0.51	2.1	0.16
908420	0.52	2.0	0.13
948660	0.44	1.8	0.14
948673	0.51	3.9	0.62
948675	0.48	2.1	0.21
948677	0.54	2.0	0.15
948951	0.45	1.9	0.11
949158	0.59	2.3	0.18
949190	0.53	2.2	0.14
949865	0.53	2.1	0.14
949873	0.47	1.8	0.20
949874	0.52	2.3	0.16
949935	0.57	2.0	0.13
950023	0.53	1.9	0.16
950030	0.59	2.1	0.20

TABLE 18

Organ weights

Compound	kidney	liver	spleen
No.	(g)	(g)	(g)

PBS	0.57	2.1	0.11
549144	0.57	2.0	0.09
950060	0.57	2.3	0.12
950089	0.60	2.4	0.15
950132	0.59	2.3	0.16
950183	0.60	2.4	0.14
951356	0.57	2.4	0.19
951362	0.58	2.3	0.16
951371	0.51	2.7	0.16
951372	0.55	2.5	0.17
951407	0.53	2.2	0.17
951431	0.54	2.2	0.18
951793	0.55	2.6	0.17
951797	0.52	2.1	0.15
951798	0.57	2.4	0.18
951799	0.54	2.4	0.16
951816	0.50	2.4	0.16
951821	0.50	2.0	0.13
951827	0.47	1.8	0.13
951839	0.51	2.2	0.15
951845	0.55	2.0	0.17
951846	0.57	2.2	0.17
951856	0.60	2.5	0.19

Claims

1. A compound comprising a modified oligonucleotide 14 to 30 linked nucleosides in length having

i) a nucleobase sequence comprising at least 14 contiguous nucleobases of SEQ ID NO: 2879;

ii) a nucleobase sequence comprising a portion of at least 14 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 87,359-90,915; 90,968-97,263; 83,573-87,287; 3,404-44,737; 97,378-104,979; 81,061-81,199; or 116,048-116,903 of SEQ ID NO: 1,

iii) a nucleobase sequence complementary to intron 12, intron 13, intron 11, intron 1, intron 14, exon 10, or the 3′-UTR of a human DNM2 pre-mRNA or human DNM2 mRNA, or

iv) a nucleobase sequence comprising at least 12, at least 13, at least 14, or at least 15 contiguous nucleobases of any one of the nucleobase sequences of SEQ ID NOs: 7-3134,

or a pharmaceutically acceptable salt thereof.

2. The compound according to claim 1, wherein the modified oligonucleotide has a nucleobase sequence consisting of any one of SEQ ID NOs: 2879, 2123, 2189, 2160, 3056, 2453, or 2232.

3. The compound according to claim 1, wherein the modified oligonucleotide comprises:

a gap segment consisting of 8-12 linked 2′-deoxynucleosides;

a 5′ wing segment consisting of 1-7 linked nucleosides; and

a 3′ wing segment consisting of 1-7 linked nucleosides;

wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each terminal wing nucleoside comprises a modified sugar.

4. The compound according to claim 1, wherein the modified oligonucleotide is at least 80%, 85%, 90%, 95% or 100% complementary to any of SEQ ID NOs: 1, 2, 3, or 3135.

5. The compound according to claim 1, wherein the modified oligonucleotide comprises at least one modified internucleoside linkage.

6. The compound according to claim 1, wherein the modified oligonucleotide comprises at least one bicyclic sugar.

7. The compound according to claim 1, wherein the modified oligonucleotide comprises at least one 5-methylcytosine.

8. The compound according to claim 1, wherein the compound is single-stranded.

9. The compound according to claim 1, wherein the compound is double-stranded.

10. The compound according to claim 1, wherein the modified oligonucleotide consists of 16 to 20 linked nucleosides.

11. A compound comprising a modified oligonucleotide according to the following formula: Gks Tks Tks Tds Ads Tds Tds Ads Tds Ads Gds Gds Gds mCks Tks Tk; wherein,

A=an adenine,

mC=a 5-methylcytosine

G=a guanine,

T=a thymine,

k=a cEt sugar moiety,

d=a 2′-deoxyribosyl sugar moiety, and

s=a phosphorothioate internucleoside linkage.

12. (canceled)

13. A compound according to the following formula:

or a salt thereof.

14. (canceled)

15. A chirally enriched population of the compound according to claim 1, wherein the population is enriched for modified oligonucleotides comprising at least one particular phosphorothioate internucleoside linkage having a particular stereochemical configuration.

16. A pharmaceutical composition comprising the compound according to claim 1 and at least one pharmaceutically acceptable diluent or carrier.

17. (canceled)

18. A method of treating, preventing, or ameliorating a disease associated with DNM2 in an individual, comprising administering to the individual the compound according to claim 1.

19. An in vitro method of inhibiting expression of DNM2 in a cell comprising contacting the cell with a single-stranded compound comprising a modified oligonucleotide 100% complementary to exon 10, an intron, or the 3′-UTR of a DNM2 nucleic acid transcript, thereby inhibiting expression of DNM2 in the cell.

20. The compound according to claim 5, wherein the at least one modified internucleoside is a phosphorothioate internucleoside linkage.

21. The compound according to claim 6, wherein the at least one bicyclic sugar is selected from the group consisting of LNA, ENA, and cEt.

22. The compound according to claim 1, comprising a conjugate group.

23. The compound according to claim 22, wherein the compound consists of the modified oligonucleotide and the conjugate group.

24. The compound according to claim 1, wherein the pharmaceutically acceptable salt is a sodium salt or a potassium salt.

25. The method according to claim 18, wherein the disease is centronuclear myopathy, Duchenne Muscular Dystrophy, Charcot-Marie Tooth disease, X-linked myotubular myopathy, autosomal recessive centronuclear myopathy, or autosomal dominant centronuclear myopathy.

26. The method according to claim 18, wherein the disease is associated with a mutation in at least one gene selected from among MTM1, BIN1, and DNM2.