EP1534728A2 - Antisense modulation of glucocorticoid receptor expression - Google Patents

Antisense modulation of glucocorticoid receptor expression

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Publication number
EP1534728A2
EP1534728A2 EP03729073A EP03729073A EP1534728A2 EP 1534728 A2 EP1534728 A2 EP 1534728A2 EP 03729073 A EP03729073 A EP 03729073A EP 03729073 A EP03729073 A EP 03729073A EP 1534728 A2 EP1534728 A2 EP 1534728A2
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Prior art keywords
seq
kcal
mol
antisense compound
acid
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EP03729073A
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German (de)
French (fr)
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EP1534728A4 (en
Inventor
Seth D. Crosby
Amy E. Nalseth
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Pharmacia LLC
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Pharmacia LLC
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Publication of EP1534728A2 publication Critical patent/EP1534728A2/en
Publication of EP1534728A4 publication Critical patent/EP1534728A4/en
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    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1138—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against receptors or cell surface proteins
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    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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Definitions

  • the present invention provides compositions and methods for modulating the expression of mammalian glucocorticoid receptor.
  • this invention relates to antisense compounds, particularly oligonucleotides, specifically hybridizable with nucleic acids encoding human glucocorticoid receptor.
  • Such oligonucleotides may modulate the expression of glucocorticoid receptor.
  • Non insulin-dependent diabetes mellitus a debilitating disease characterized by abnormal elevation of blood glucose levels, is driven by three factors: increased hepatic glucose production, inadequate clearance of glucose via insulin mediated pathways, and decreased uptake of circulating glucose by tissues. (Diabetes Review 5(3), 177-269, (1997)). Administration of agents which decrease hepatic glucose production are a fundamental approach to controlling blood glucose levels (Am. J. Physiol. 257, E35-E42 (1989). J. Clin. Endocrinol Metab. 11, 1 180-1183 (1994) and J Clin. Invest., 92, 2283-2290 (1993)).
  • Glucocorticoids have been shown to have major influences on glucose production. Glucocorticoid excess aggravates established diabetes and glucocorticoid deficiency reduces blood glucose and improves glucose control in diabetic mice. (Diabetes Review, 1(3), 301-308, (1993). Diabetologia, 9, 376-379 (1973). Horm. Metab. Res., 9, 152-156 (1977)). The underlying mechanism responsible for these effects is believed to be glucocorticoid-induced upregulation of hepatic enzymes required for gluconeogenesis. (Recent Prog. Horm. Res., 26, 41 1-457 (1970). Physiol. Rev., 64, 170-259 (1984).
  • the glucocorticoids are lipid soluble hormones synthesized in the adrenal cortex. They readily pass through cell membranes, enter the cytoplasm of target tissues, and bind to glucocorticoid receptors in the cytoplasm by complexation with heat shock proteins. Upon binding of the hormone to its receptor, the receptor undergoes a conformational change which results in dissociation of heat shock proteins and translocation of the ligand-bound glucocorticoid receptor into the nucleus where it can either initiate or repress specific gene transcription.
  • Transcriptional activation occurs when the ligand bound receptor complex homodimerizes, and the homodimeric receptor ligand complex binds to chromosomal DNA at sequence-specific sites in the promoter region of regulated genes.
  • genes which glucocorticoids up-regulate are those which play key roles in gluconeogenesis and glycogenolysis, particularly PEPCK and glucose-6- phosphatase. (Advanced Enzymology., Meister, Ed. New York, John Wiley and Sons, Inc., 203-281 (1994). and Diabetes 45, 1563-1571(1996)).
  • Phosphoenolpyruvatecarboxy kinase catalyzes the conversion of oxaloacetate to phosphoenolpyruvate and glucose-6- phosphatase catalyzes the conversion of glucose-6-phosphate into glucose, both of which are required for the synthesis of glucose from oxaloacetate in the liver.
  • Aventis® a potent glucocorticoid receptor antagonist, reduces mRNA levels of PEPCK and glucose-6-phosphate in the liver and causes a 50% reduction of plasma glucose levels in obese diabetic db/db transgenic mice.
  • agents which function as glucocorticoid antagonists represent a novel approach to controlling type II diabetes
  • agents which antagonize the glucocorticoid receptor have been the subject of active current research for their clinical potential.
  • U.S. Pat. No. 5,929,058 discloses a method for treating type II diabetes comprising administering a combination of nonselective steroidal agents exhibiting mineralcorticoid receptor agonist activity and glucocorticoid receptor antagonist activity.
  • non- steroidal, glucocorticoid-selective agents which antagonize the glucocorticoid receptor. These compounds would be particularly useful for treating type II diabetes and the symptoms thereof, such as hyperglycemia, inadequate glucose clearance, obesity, hyperinsulinemia, hypertriglyceridemia, and high circulating glucocorticoid levels.
  • Antisense technology is emerging as an effective means for reducing the expression of specific gene products and may therefore prove to be uniquely useful in a number of therapeutic, diagnostic, and research applications for the modulation of GR expression.
  • Figure 1 shows the cDNA sequence of human glucocorticoid receptor and the encoded protein sequence.
  • Figure 2 shows the cDNA sequence of murine glucocorticoid receptor and the encoded protein sequence.
  • the present invention is directed to antisense compounds, particularly oligonucleotides, which are targeted to a nucleic acid encoding glucocorticoid receptor, and which modulate the expression of mammalian glucocorticoid receptor (GR).
  • Pharmaceutical and other compositions comprising the antisense compounds of the invention are also provided. Further provided are methods of modulating the expression of glucocorticoid receptor in cells or tissues comprising contacting said cells or tissues with one or more of the antisense compounds or compositions of the invention.
  • the present invention employs oligomeric antisense compounds, particularly oligonucleotides, for use in modulating the function of nucleic acid molecules encoding glucocorticoid receptor, ultimately modulating the amount of glucocorticoid receptor produced. This is accomplished by providing antisense compounds, which specifically hybridize with one or more nucleic acids encoding glucocorticoid receptor.
  • target nucleic acid and “nucleic acid encoding GR” encompass DNA encoding GR, RNA (including pre-mRNA and mRNA) transcribed from such DNA, and also cDNA derived from such RNA.
  • the specific hybridization of an oligomeric compound with its target nucleic acid interferes with the normal function of the nucleic acid.
  • This modulation of function of a target nucleic acid by compounds, which specifically hybridize to it, is generally referred to as "antisense".
  • the functions of DNA to be interfered with include replication and transcription.
  • the functions of RNA to be interfered with include all vital functions such as, for example, translocation of the RNA to the site of protein translation, translation of protein from the RNA, splicing of the RNA to yield one or more mRNA species, and catalytic activity which may be engaged in or facilitated by the RNA.
  • the overall effect of such interference with target nucleic acid function is modulation of the expression of GR.
  • modulation means either an increase (stimulation) or a decrease (inhibition) in the expression of a gene.
  • inhibition is the preferred form of modulation, of gene expression and mRNA is a preferred target.
  • Targeting an antisense compound to a particular nucleic acid is a multistep process. The process usually begins with the identification of a nucleic acid sequence whose function is to be modulated. This may be, for example, a cellular gene (or mRNA transcribed from the gene) whose expression is associated with a particular disorder or disease state, or a nucleic acid molecule from an infectious agent. In the present invention, the target is a nucleic acid molecule encoding GR.
  • the targeting process also includes determination of a site or sites within this gene for the antisense interaction to occur such that the desired effect, e.g., detection or modulation of expression of the protein, will result.
  • a preferred intragenic site is the region encompassing the translation initiation or termination codon of the open reading frame (ORF) of the gene. Since, as is known in the art, the translation initiation codon is typically 5'-AUG (in transcribed mRNA molecules; 5'-ATG in the corresponding DNA molecule), the translation initiation codon is also referred to as the "AUG codon,” the "start codon” or the "AUG start codon".
  • translation initiation codon having the RNA sequence 5'-GUG, 5'-UUG or 5'-CUG, and 5'-AUA, 5'-ACG and 5'-CUG have been shown to function in vivo.
  • the terms "translation initiation codon” and "start codon” can encompass many codon sequences, even though the initiator amino acid in each instance is typically methionine (in eukaryotes) or formyl methionine (in prokaryotes). It is also known in the art that eukaryotic and prokaryotic genes may have two or more alternative start codons, any one of which may be preferentially utilized for translation initiation in a particular cell type or tissue, or under a particular set of conditions.
  • start codon and “translation initiation codon” refer to the codon or codons that are used in vivo to initiate translation of an mRNA molecule transcribed from a gene encoding GR, regardless of the sequence(s) of such codons.
  • a translation termination codon (or "stop codon”) of a gene may have one of three sequences, i.e. 5'-UAA, 5'-UAG and 5'- UGA (the corresponding DNA sequences are 5'-TAA, 5'-TAG and 5'-TGA, respectively).
  • start codon region and “translation initiation codon region” “refer to a portion of such an mRNA or gene that encompasses from about 25 to about 50 contiguous nucleotides in either direction (i.e., 5' or 3') from a translation initiation codon.
  • stop codon region and “translation termination codon region” refer to a portion of such an mRNA or gene that encompasses from about 25 to about 50 contiguous nucleotides in either direction (i.e., 5' or 3') from a translation termination codon.
  • target regions include the 5' untranslated region (5 'UTR), known in the art to refer to the portion of an mRNA in the 5' direction from the translation initiation codon, and thus including nucleotides between the 5' cap site and the translation initiation codon of an mRNA or corresponding nucleotides on the gene, and the 3' untranslated region (3 'UTR), known in the art to refer to the portion of an mRNA in the 3' direction from the translation termination codon, and thus including nucleotides between the translation termination codon and 3' end of an mRNA or corresponding nucleotides on the gene.
  • 5 'UTR 5' untranslated region
  • 3 'UTR known in the art to refer to the portion of an mRNA in the 3' direction from the translation termination codon, and thus including nucleotides between the translation termination codon and 3' end of an mRNA or corresponding nucleotides on the gene.
  • the 5' cap of an mRNA comprises an N7- methylated guanosine residue joined to the 5 '-most residue of the mRNA via a 5 '-5' triphosphate linkage.
  • the 5' cap region of an mRNA is considered to include the 5' cap structure itself as well as the first 50 nucleotides adjacent to the cap.
  • the 5' cap region may also be a preferred target region.
  • mRNA splice sites i.e., intron-exon junctions
  • intron-exon junctions may also be preferred target regions, and are particularly useful in situations where aberrant splicing is implicated in disease, or where an overproduction of a particular mRNA splice product is implicated in disease. Aberrant fusion junctions due to rearrangements or deletions are also preferred targets.
  • introns can also be effective, and therefore preferred, target regions for antisense compounds targeted, for example, to DNA or pre-mRNA.
  • oligonucleotides are chosen which are sufficiently complementary to the target, i.e., hybridize sufficiently well and with sufficient specificity, to give the desired effect.
  • hybridization means hydrogen bonding, which may be Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding, between complementary nucleoside or nucleotide bases.
  • adenine and thymine are complementary nucleobases, which pair through the formation of hydrogen bonds.
  • “Complementary,” as used herein, refers to the capacity for precise pairing between two nucleotides. For example, if a nucleotide at a certain position of an oligonucleotide is capable of hydrogen bonding with a nucleotide at the same position of a DNA or RNA molecule, then the oligonucleotide and the DNA or RNA are considered to be complementary to each other at that position. The oligonucleotide and the DNA or RNA are complementary to each other when a sufficient number of corresponding positions in each molecule are occupied by nucleotides which can hydrogen bond with each other.
  • An antisense compound is specifically hybridizable when binding of the compound to the target DNA or RNA molecule interferes with the normal function of the target DNA or RNA to cause a loss of utility, and there is a sufficient degree of complementarity to avoid non-specific binding of the antisense compound to non-target sequences under conditions in which specific binding is desired, i.e., under physiological conditions in the case of in vivo assays or therapeutic treatment, and in the case of in vitro assays, under conditions in which the assays are performed.
  • Antisense compounds are commonly used as research reagents and diagnostics.
  • antisense oligonucleotides which are able to inhibit gene expression with extraordinar specificity, are often used by those of ordinary skill to elucidate the function of particular genes. Antisense compounds are also used, for example, to distinguish between functions of various members of a biological pathway. Antisense modulation has, therefore, been harnessed for research use. [0020] The specificity and sensitivity of antisense is also harnessed by those of skill in the art for therapeutic uses. Antisense oligonucleotides have been employed as therapeutic moieties in the treatment of disease states in animals and man.
  • oligonucleotide refers to an oligomer or polymer of ribonucleic acid (RNA) or deoxyribonucleic acid (DNA) or mimetics thereof. This term includes oligonucleotides composed of naturally occurring nucleobases, sugars and covalent internucleoside (backbone) linkages as well as oligonucleotides having non-naturally occurring portions which function similarly.
  • modified or substituted oligonucleotides are often preferred over native forms because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for nucleic acid target and increased stability in the presence of nucleases.
  • antisense oligonucleotides are a preferred form of antisense compound, the present invention comprehends other oligomeric antisense compounds, including but not limited to oligonuclectide mimetics such as are described below.
  • the antisense compounds in accordance with this invention preferably comprise 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, or 50 nucleobases.
  • the antisense compounds in accordance with this invention preferably comprise from about 8 to about 30 nucleobases (i.e. from about 8 to about 30 linked nucleo sides).
  • Particularly preferred antisense compounds are antisense oligonuclectides, even more preferably those comprising from about 12 to about 25 nucleobases.
  • the antisense compounds in accordance with this invention preferably comprise a nucleic acid sequence selected from the group consisiting of SEQ ID NO: 1 -SEQ ID NO:4769, and a fragment of at least eight contiguous nucleotides of SEQ ID NO: 1 -SEQ ID NO:4769.
  • a nucleoside is a base-sugar combination. The base portion of the nucleoside is normally a heterocyclic base. The two most common classes of such heterocyclic bases are the purines and the pyrimidines.
  • Nucleotides are nucleosides that further include a phosphate group covalently linked to the sugar portion of the nucleoside.
  • the phosphate group can be linked to either the 2', 3' or 5' hydroxyl moiety of the sugar.
  • the phosphate groups covalently link adjacent nucleosides to one another to form a linear polymeric compound.
  • the respective ends of this linear polymeric structure can be further joined to form a circular structure, however, open linear structures are generally preferred.
  • the phosphate groups are commonly referred to as forming the internucleoside backbone of the oligonucleotide.
  • the normal I linkage or backbone of RNA and DNA is a 3' to 5' phosphodiester linkage.
  • oligonucleotides containing modified backbones or non-natural internucleoside linkages include those that retain a phosphorus atom in the backbone and those that do not have a phosphorus atom in the backbone.
  • modified oligonucleotides that do not have a phosphorus atom in their internucleoside backbone can also be considered to be oligonucleosides.
  • Preferred modified oligonucleotide backbones include, for example, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkylphosphotriesters, methyl and other alkyl phosphonates including 3 'alkylene phosphonates and chiral phosphonates, phosphinates, phosphoramidates including 3 '-amino phosphoramidate and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, and boranophosphates having normal 3'-5' linkages, 2'-5' linked analogs of these, and those having inverted polarity wherein the adjacent pairs of nucleoside units are linked 3'-5' to 5'-3' or 2'-5' to 5'-2'.
  • Various salts, mixed salts and free acid forms are also included.
  • Representative United States patents that teach the preparation of the above phosphorus-containing linkages include, but are not limited to, U.S.: 3,687,808; 4,469,863; 4,476,301 ; 5,023,243; 5,177,196; 5,188,897; 5,264,423; 5,276,019; 5,278,302; 5,286,717; 5,321,131 ; 5,399,676; 5,405,939; 5,453,496; 5,455,233; 5,466,677; 5,476,925; 5,519,126; 5,536,821; 5,541 ,306; 5,550,1 1 1; 5,563,253; 5,571,799; 5,587,361 ; and 5,625,050, each of which is herein inco ⁇ orated by reference.
  • Preferred modified oligonucleotide backbones that do not include a phosphorus atom therein have backbones that are formed by short chain alkyl or cycloalkyl internucleoside linkages, mixed heteroatom and alkyl or cycloalkyl internucleoside linkages, or one or more short chain heteroatomic or heterocyclic internucleoside linkages.
  • morpholino linkages formed in part from the sugar portion of a nucleoside
  • siloxane backbones sulfide, sulfoxide and sulfone backbones
  • formacetyl and thioformacetyl backbones methylene formacetyl and thioformacetyl backbones
  • alkene containing backbones sulfamate backbones
  • sulfonate and sulfonamide backbones amide backbones; and others having mixed N, O, S and CH component parts.
  • Representative United States patents that teach the preparation of the above oligonucleosides include, but are not limited to, U.S.: 5,034,506; 5,166,315; 5,185,444; 5,214,134; 5,216,141; 5,235,033; 5,264,562; 5,264,564; 5,405,938; 5,434,257; 5,466,677; 5,470,967; 5,489,677; 5,541 ,307; 5,561,225; 5,596,086; 5,602,240; 5,610,289; 5,602,240; 5,608,046; 5,610,289; 5,618,704; 5,623,070; 5,663,312; 5,633,360; 5,677,437; and 5,677,439, each of which is herein incorporated by reference.
  • both the sugar and the internucleoside linkage, i.e., the backbone, of the nucleotide units are replaced with novel groups.
  • the base units are maintained for hybridization with an appropriate nucleic acid target compound, one such oligomeric compound, an oligonucleotide mimetic that has been shown to have excellent hybridization properties, is referred to as a peptide nucleic acid (PNA).
  • PNA peptide nucleic acid
  • the sugar-backbone of an oligonucleotide is replaced with an amide containing backbone, in particular an aminoethylglycine backbone.
  • nucleobases are retained and are bound directly or indirectly to aza nitrogen atoms of the amide portion of the backbone.
  • Representative United States patents that teach the preparation of PNA compounds include, but are not limited to, U.S.: 5,539,082; 5,714,331 ; and 5,719,262, each of which is herein incorporated by reference. Further teaching of PNA compounds can be found in Nielsen et al., Science, 1991 , 254, 1497-1500.
  • Most preferred embodiments of the invention are oligonucleotides with phosphorothioate backbones and oligonucleosides with heteroatom backbones, and in particular -CH 2 -NH-O-CH 2 -, -CH 2 -N (CH 3 ) -O-CH 2 - [known as a methylene (methylimino) or MMI backbone] , - CH 2 -O-N (CH 3 ) -CH 2 -, -CH 2 N(CH 3 )-N(CH 3 )- CH 2 - and -O-N(CH 3 )-CH -CH2- [wherein the native phosphodiester backbone is represented as -O-P-O-CH 2 -] of the above referenced U.S.
  • Modified oligonucleotides may also contain one or more substituted sugar moieties.
  • Preferred oligonucleotides comprise one of the following at the 2' position: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S- or N-alkynyl; or O- alkyl-O-alkyl, wherein the alkyl, alkenyl and alkynyl may be substituted or unsubstituted C ⁇ to C ⁇ 0 alkyl or C 2 to Cio alkenyl and alkynyl.
  • oligonucleotides comprise one of the following at the 2' position: Ci to Cio, ( lower alkyl, substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH 3 , OCN, Cl, Br, CN, CF 3 , OCF 3 , SOCH 3 , SO 2 CH 3 , ON0 2 , NO 2 , N 3 , NH 2 , heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, an RNA cleaving group, a reporter group, an intercalator, a group for improving the pharmacokinetic properties of an oligonucleotide, or a group for improving the pharmacodynamic properties of an oligonucleotide, and other substituents having similar properties.
  • a preferred modification includes 2' - methoxyethoxy (2' -O-CH 2 CH 2 OCH 3 , also known as 2'-O- (2-methoxyethyl) or 2'- MOE) (Martin et al., Helv. Chim. Acta, 1995, 78, 486-504) i.e., an alkoxyalkoxy group.
  • a further preferred modification includes 2'-dimethylaminooxyethoxy, i.e., a O(CH 2 ) 2 ON(CH ) 2 group, also known as 2'-DMAOE, as described in examples herein below, and 2'-dimethylaminoethoxyethoxy (also known in the art as 2'-O- dimethyla inoethoxyethyl or 2'-DMAEOE), i.e., 2'-O-CH 2 -O-CH.-N (CH 2 ) 2 , also described in examples herein below.
  • Oligonucleotides may also include nucleobase (often referred to in the art simply as “base”) modifications or substitutions.
  • nucleobases include the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C) and uracil (U).
  • Modified nucleobases include other synthetic and natural nucleobases such as 5- methylcytosine (5-me-C), 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2- aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2- propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2- thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo particularly 5-bromo, 5-trifluoromethyl and other 5-substitute
  • nucleobases include those disclosed in United States Patent No. 3,687,808, those disclosed in The Concise Encyclopedia Of Polymer Science And Engineering, pages 858-859, Kroschwitz, J.I., ed. John Wiley & Sons, 1990, those disclosed by Englisch et al., Angewandte Chemie, International Edition, 1991 , 30, 613, and those disclosed by Sanghvi, Y.S., Chapter 15, Antisense Research and Applications, pages 289-302, Crooke, S.T. and Lebleu, B. ed., CRC Press, 1993. Certain of these nucleobases are particularly useful for increasing the binding affinity of the oligomeric compounds of the invention.
  • 5-substituted pyrimidines include 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and O-6 substituted purines, including 2-aminopropyladenine, 5-propynyluracil and 5-propynylcytosine.
  • 5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2°C (Sanghvi, Y.S., Crooke, S.T. and Lebleu, B., eds, Antisense Research and Applications, CRC Press, Boca Raton, 1993, pp. 276-278) and are presently preferred base substitutions, even more particularly when combined with 2'-O- methoxyethyl sugar modifications.
  • oligonucleotides of the invention involves chemically linking to the oligonucleotide one or more moieties or conjugates, which enhance the activity, cellular distribution or cellular uptake of the oligonucleotide.
  • moieties include but are not limited to lipid moieties such as a cholesterol moiety (Letsinger et al., Proc. Natl. Acad. Sci. USA, 1989, 86, 6553-6556), cholic acid (Manoharan et al., Bioorg. Med. Chem.
  • a thioether e.g., hexyl-S-tritylthiol (Manoharan et a ⁇ ., Ann. N. Y. Ac ⁇ d. Sci., 1992, 660, 306-309; Manoharan et al., Bioorg. Med. Chem. Let., 1993, 3, 2765-2770), a thiocholesterol (Oberhauser et al., Nucl.
  • Acids Res., 1990, 18, 3777-3783 a polyamine or a polyethylene glycol chain (Mancharan et al., Nucleosides & Nucleotides, 1995, 14, 969-973), or adamantane acetic acid (Manoharan et al., Tetrahedron Lett., 1995, 36, 365'-3654), a palmityl moiety (Mishra et al., Biochim. Biophys. Acta, 1995, 1264, 229-237), or an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety (Crooke et al., J Pharmacol. Exp. Ther., 1996, 277, 923-937).
  • Representative United States patents that teach the preparation of such oligonucleotide conjugates include, but are not limited to, U.S.: 4,828,979; 4,948,882; 5,218,105; 5,525,465; 5,541 ,313; 5,545,730; 5,552,538; 5,578,717, 5,580,731; 5,580,731 ; 5,591 ,584; 5,109,124; 5,1 18,802; 5,138,045; 5,414,077; 5,486,603; 5,512,439; 5,578,718; 5,608,046; 4,587,044; 4,605,735; 4,667,025; 4,762,779; 4,789,737; 4,824,941 ; 4,835,263; 4,876,335; 4,904,582; 4,958,013; 5,082,830; 5,112,963; 5,214,136; 5,082,830; 5,112,963; 5,214,136; 5,24
  • antisense compounds which are chimeric compounds.
  • Chimeric antisense compounds or “chimeras,” in the context of this invention are antisense compounds, particularly oligonucleotides, which contain two or more chemically distinct regions, each made up of at least one monomer unit, i.e., a nucleotide in the case of an oligonucleotide compound.
  • oligonucleotides typically contain at least one region wherein the oligonucleotide is modified so as to confer upon the oligonucleotide increased resistance to nuclease degradation, increased cellular uptake, and/or increased binding affinity for the target nucleic acid.
  • An additional region of the oligonucleotide may serve as a substrate for enzymes capable of cleaving RNA:DNA or RNA:RNA hybrids.
  • RNase H is a cellular endonuclease, which cleaves the RNA strand of RNA:DNA duplex.
  • RNA target Activation of RNase H, therefore, results in cleavage of the RNA target, thereby greatly enhancing the efficiency of oligonucleotide inhibition of gene expression. Consequently, comparable results can often be obtained with shorter oligonucleotides when chimeric oligonucleotides are used, compared to phosphorothioate deoxyoligonucleotides hybridizing to the same target region.
  • Cleavage of the RNA target can be routinely detected by gel electrophoresis and, if necessary, associated nucleic acid hybridization techniques known in the art.
  • Chimeric antisense compounds of the invention may be formed as composite structures of two or more oligonucleotides, modified oligonucleotides, oligonucleosides and/or oligonucleotide mimetics as described above. Such compounds have also been referred to in the art as hybrids or gapmers.
  • the antisense compounds used in accordance with this invention may be conveniently, and routinely made through the well-known technique of solid phase synthesis.
  • Equipment for such synthesis is sold by several vendors including, for example, Applied Biosystems (Foster City, CA). Any other means for such synthesis known in the art may additionally or alternatively be employed. It is well known to use similar techniques to prepare oligonucleotides such as the phosphorothioates and alkylated derivatives.
  • the antisense compounds of the invention are synthesized in vitro and do not include antisense compositions of biological origin, or genetic vector constructs designed to direct the in vivo synthesis of antisense molecules.
  • the compounds of the invention may also be admixed, encapsulated, conjugated or otherwise associated with other molecules, molecule structures or mixtures of compounds, as for example, liposomes, receptor targeted molecules, oral, rectal, topical or other formulations, for assisting in uptake, distribution and/or abso ⁇ tion.
  • Representative United States patents that teach the preparation of such uptake, distribution and/or abso ⁇ tion assisting formulations include, but are not limited to, U.S.: 5,108,921 ; 5,354,844; 5,416,016; 5,459,127; 5,521 ,291 ; 5,543,158; 5,547,932; 5,583,020; 5,591 ,721 ; 4,426,330; 4,534,899; 5,013,556; 5,108,921 ; 5,213,804; 5,227,170; 5,264,221 ; 5,356,633; 5,395,619; 5,416,016; 5,417,978; 5,462,854; 5,469,854; 5,512,295; 5,527,528; 5,534,259; 5,543,152; 5,556,948; 5,580,575; and 5,595,756, each of which is herein inco ⁇ orated by reference.
  • the antisense compounds of the invention encompass any pharmaceutically acceptable salts, esters, or salts of such esters, or any other compound which, upon administration to an animal including a human, is capable of providing (directly or indirectly) the biologically active metabolite or residue thereof. Accordingly, for example, the disclosure is also drawn to prodrugs and pharmaceutically acceptable salts of the compounds of the invention, pharmaceutically acceptable salts of such prodrugs, and other bioequivalents. [0040]
  • prodrug indicates a therapeutic agent that is prepared in an inactive form that is converted to an active form (i.e., drug) within the body or cells thereof by the action of endogenous enzymes or other chemicals and/or conditions.
  • prodrug versions of the oligonuclectides of the invention are prepared as SATE [(S-acetyl-2-thioethyl) phosphate] derivatives according to the methods disclosed in WO 93/24510 to Gosselin et al., published December 9, 1993 or in WO 94/26764 to Imbach et al.
  • pharmaceutically acceptable salts refers to physiologically and pharmaceutically acceptable salts of the compounds of the invention: i.e., salts that retain the desired biological activity of the parent compound and do not impart undesired toxicological effects thereto.
  • Pharmaceutically acceptable base addition salts are formed with metals or amines, such as alkali and alkaline earth metals or organic amines.
  • metals used as cations are sodium, potassium, magnesium, calcium, and the like.
  • suitable amines are N, N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, dicyclohexylamine, ethylenediamine, N-methylglucamine, and procaine (see, for example, Berge et al., "Pharmaceutical Salts," J. of Pharma Sci., 1977, 66, 119).
  • the base addition salts of said acidic compounds are prepared by contacting the free acid form with a sufficient amount of the desired base to produce the salt in the conventional manner.
  • the free acid form may be regenerated by contacting the salt form with an acid and isolating the free acid in the conventional manner.
  • the free acid forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free acid for pu ⁇ oses of the present invention.
  • a "pharmaceutical addition salt” includes a pharmaceutically acceptable salt of an acid form of one of the components of the compositions of the invention. These include organic or inorganic acid salts of the amines.
  • Preferred acid salts are the hydrochlorides, acetates, salicylates, nitrates and phosphates.
  • Other suitable pharmaceutically acceptable salts are well known to those skilled in the art and include basic salts of a variety of inorganic and organic acids, such as, for example, with inorganic acids, such as for example hydrochloric acid, hydrobromic acid, sulfuric acid or phosphoric acid; with organic carboxylic, sulfonic, sulfo or phospho acids or N-substituted sulfamic acids, for example acetic acid, propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid, methylmaleic acid, fumaric acid, malic acid, tartaric acid, lactic acid, oxalic acid, gluconic acid, glucaric acid, glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-aminosalicylic
  • Pharmaceutically acceptable salts of compounds may also be prepared with a pharmaceutically acceptable cation.
  • Suitable pharmaceutically acceptable cations are well known to those skilled in the art and include alkaline, alkaline earth, ammonium and quaternary ammonium cations. Carbonates or hydrogen carbonates are also possible.
  • salts formed with cations such as sodium, potassium, ammonium, magnesium, calcium, polyamines such as spermine and spermidine, etc.
  • acid addition salts formed with inorganic acids for example hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid and the like
  • salts formed with organic acids such as, for example, acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, methanesulfonic acid, p- toluenesulfonic acid, naphthalenedisulfonic acid, polygal
  • the antisense compounds of the present invention can be utilized for diagnostics, therapeutics, prophylaxis and as research reagents and kits.
  • an animal preferably a human, suspected of having a disease or disorder, which can be treated by modulating the expression of GR, is treated by administering antisense compounds in accordance with this invention.
  • the compounds of the invention can be utilized in pharmaceutical compositions by adding an effective amount of an antisense compound to a suitable pharmaceutically acceptable diluent or carrier.
  • Use of the antisense compounds and methods of the invention may also be useful prophylactically, e.g., to prevent or delay infection, inflammation or tumor formation, for example.
  • the antisense compounds of the invention are useful for research and diagnostics, because these compounds hybridize to nucleic acids encoding GR, enabling sandwich and other assays to easily be constructed to exploit this fact.
  • Hybridization of the antisense oligonucleotides of the invention with a nucleic acid encoding GR can be detected by means known in the art. Such means may include conjugation of an enzyme to the oligonucleotide, radiolabelling of the oligonucleotide or any other suitable detection means. Kits using such detection means for detecting the level of GR in a sample may also be prepared.
  • the present invention also includes pharmaceutical compositions and formulations, which include the antisense compounds of the invention.
  • compositions of the present invention may be administered in a number of ways depending upon whether local or systemic treatment is desired and upon the area to be treated. Administration may be topical (including ophthalmic and to mucous membranes including vaginal and rectal delivery), pulmonary, e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, intranasal, epidermal and transdermal), oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial, e.g., intrathecal or intraventricular, administration. Oligonucleotides with at least one 2'-O- methoxyethyl modification are believed to be particularly useful for oral administration.
  • compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders.
  • Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.
  • Coated condoms, gloves and the like may also be useful.
  • Compositions and formulations for oral administration include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets or tablets. Thickeners, flavoring agents, diluents, emulsifiers, dispersing aids or binders may be desirable.
  • compositions and formulations for parenteral, intrathecal or intraventricular administration may include sterile aqueous solutions, which may also contain buffers, diluents and other suitable additives such as, but not limited to, penetration enhancers, carrier compounds and other pharmaceutically acceptable carriers or excipients.
  • compositions of the present invention include, but are not limited to, solutions, emulsions, and liposome-containing formulations. These compositions may be generated from a variety of components that include, but are not limited to, preformed liquids, self-emulsifying solids and self-emulsifying semisolids.
  • compositions of the present invention may be prepared according to conventional techniques well known in the pharmaceutical industry. Such techniques include the step of bringing into association the active ingredients with the pharmaceutical carrier(s) or excipient(s). In general the formulations are prepared by uniformly and intimately bringing into association the active ingredients with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.
  • compositions of the present invention may be formulated into any of many possible dosage forms such as, but not limited to, tablets, capsules, liquid syrups, soft gels, suppositories, and enemas.
  • compositions of the present invention may also be formulated as suspensions in aqueous, non-aqueous or mixed media.
  • Aqueous suspensions may further contain substances, which increase the viscosity of the suspension including, for example, sodium carboxymethylcellulose, sorbitol and/or dextran.
  • the suspension may also contain stabilizers.
  • the pharmaceutical compositions may be formulated and used as foams.
  • Pharmaceutical foams include formulations such as, but not limited to, emulsions, microemulsions, creams, jellies and liposomes. While basically similar in nature these formulations vary in the components and the consistency of the final product.
  • the preparation of such compositions and formulations is generally known to those skilled in the pharmaceutical and formulation arts and may be applied to the formulation of the compositions of the present invention. Emulsions
  • compositions of the present invention may be prepared and formulated as emulsions.
  • Emulsions are typically heterogenous systems of one liquid dispersed in another in the form of droplets usually exceeding 0.1 ⁇ m in diameter.
  • Emulsions are often biphasic systems comprising of two immiscible liquid phases intimately mixed and dispersed with each other.
  • emulsions may be either water-in-oil (w/o) or of the oil-in-water (o/w) variety.
  • Emulsions may contain additional components in addition to the dispersed phases and the active drug, which may be present as a solution in either the aqueous phase, oily phase or itself as a separate phase. Pharmaceutical excipients such as emulsifiers, stabilizers, dyes, and anti-oxidants may also be present in emulsions as needed.
  • compositions may also be multiple emulsions that are comprised of more than two phases such as, for example, in the case of oil-in-water-in-oil (o/w/o) and water-in- oil-in-water (w/o/w) emulsions.
  • Such complex formulations often provide certain advantages that simple binary emulsions do not.
  • Multiple emulsions in which individual oil droplets of an o/w emulsion enclose small water droplets constitute a w/o/w emulsion.
  • a system of oil droplets enclosed in globules of water stabilized in an oily continuous provides an o/w/o emulsion.
  • Emulsions are characterized by little or no thermodynamic stability. Often, the dispersed or discontinuous phase of the emulsion is well dispersed into the external or continuous phase and maintained in this form through the means of emulsifiers or the viscosity of the formulation. Either of the phases of the emulsion may be a semisolid or a solid, as is the case of emulsion-style ointment bases and creams. Other means of stabilizing emulsions entail the use of emulsifiers that may be incorporated into either phase of the emulsion.
  • Emulsifiers may broadly be classified into four categories: synthetic surfactants, naturally occurring emulsifiers, abso ⁇ tion bases, and finely dispersed solids (Idson, in Pharmaceutical Dosaqe Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199).
  • Synthetic surfactants also known as surface active agents, have found wide applicability in the formulation of emulsions and have been reviewed in the literature (Rieger, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 285; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), Marcel Dekker, Inc., New York, N.Y., 1988, volume 1, p. 199).
  • Surfactants are typically amphiphilic and comprise a hydrophilic and a hydrophobic portion.
  • HLB hydrophile/lipophile balance
  • surfactants may be classified into different classes based on the nature of the hydrophilic group: nonionic, anionic, cationic and amphoteric (Rieger, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1 , p. 285).
  • Naturally occurring emulsifiers used in emulsion formulations include lanolin, beeswax, phosphatides, lecithin and acacia. Abso ⁇ tion bases possess hydrophilic properties such that they can soak up water to form w/o emulsions yet retain their semisolid consistencies, such as anhydrous lanolin and hydrophilic petrolatum. Finely divided solids have also been used as good emulsifiers especially in combination with surfactants and in viscous preparations.
  • polar inorganic solids such as heavy metal hydroxides, nonswelling clays such as bentonite, attapulgite, hectorite, kaolin, montmorillonite, colloidal aluminum silicate and colloidal magnesium aluminum silicate, pigments and nonpolar solids such as carbon or glyceryl tristearate.
  • non-emulsifying materials are also included in emulsion formulations and contribute to the properties of emulsions.
  • Hydrophilic colloids or hydrocolloids include naturally occurring gums and synthetic polymers such as polysaccharides (for example, acacia, agar, alginic acid, carrageenan, guar gum, karaya gum, and tragacanth), cellulose derivatives (for example, carboxymethyl cellulose and carboxypropylcellulose), and synthetic polymers (for example, carbomers, cellulose ethers, and carboxyvinyl polymers). These disperse or swell in water to form colloidal solutions that stabilize emulsions by forming strong interfacial films around the dispersedphase droplets and by increasing the viscosity of the external phase.
  • polysaccharides for example, acacia, agar, alginic acid, carrageenan, guar gum, karaya gum, and tragacanth
  • cellulose derivatives for example, carboxymethyl cellulose and carboxypropylcellulose
  • synthetic polymers for example, carbomers, cellulose ethers,
  • emulsions often contain a number of ingredients such as carbohydrates, proteins, sterols and phosphatides that may readily support the growth of microbes, these formulations often inco ⁇ orate preservatives.
  • preservatives included in emulsion formulations include methyl paraben, propyl paraben, quaternary ammonium salts, benzalkonium chloride, esters of p- hydroxybenzoic acid, and boric acid.
  • Antioxidants are also commonly added to emulsion formulations to prevent deterioration of the formulation.
  • Antioxidants used may be free radical scavengers such as tocopherols, alkyl gallates, butylated hydroxyanisole, butylated hydroxytoluene, or reducing agents such as ascorbic acid and sodium metabisulfite, and antioxidant synergists such as citric acid, tartaric acid, and lecithin.
  • free radical scavengers such as tocopherols, alkyl gallates, butylated hydroxyanisole, butylated hydroxytoluene, or reducing agents such as ascorbic acid and sodium metabisulfite
  • antioxidant synergists such as citric acid, tartaric acid, and lecithin.
  • compositions of oligonucleotides and nucleic acids are formulated as microemulsions.
  • a microemulsion may be defined as a system of water, oil and amphiphile, which is a single optically isotropic, and thermodynamically stable liquid solution (Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1 , p. 245).
  • microemulsions are systems that are prepared by first dispersing an oil in an aqueous surfactant solution and then adding a sufficient amount of a fourth component, generally an intermediate chain-length alcohol to form a transparent system.
  • microemulsions have also been described as thermodynamically stable, isotropically clear dispersions of two immiscible liquids that are stabilized by interfacial films of surface-active molecules (Leung and Shah, in: Controlled Release of Drugs: Polymers and Aggregate Systems, Rosoff, M., Ed., 1989, VCH Publishers, New York, pages 1852 '5).
  • Microemulsions commonly are prepared via a combination of three to five components that include oil, water, surfactant, cosurfactant and electrolyte.
  • microemulsion is of the water-in-oil (w/o) or an oil-in-water (o/w) type is dependent on the properties of the oil and surfactant used and on the structure and geometric packing of the polar heads and hydrocarbon tails of the surfactant molecules (Schott, in Remington 's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA, 1985, p. 271).
  • microemulsions offer the advantage of solubilizing water-insoluble drugs in a formulation of thermodynamically stable droplets that are formed spontaneously.
  • Surfactants used in the preparation of microemulsions include, but are not limited to, ionic surfactants, non-ionic surfactants, Brij 96, polyoxyethylene oleyl ethers, polyglycerol fatty acid esters, tetraglycerol monolaurate (ML310), tetraglycerol monooleate (MO310), hexaglycerol monooleate (PO310), hexaglycerol pentaoleate (PO500), decaglycerol monocaprate (MCA750), decaglycerol monooleate (MO750), decaglycerol sequioleate (S0750), decaglycerol decaoleate (DAO750), alone or in combination with cosurfactants.
  • the cosurfactant usually a short-chain alcohol such as ethanol, 1-propanol, and 1- butanol, serves to increase the interfacial fluidity by penetrating into the surfactant film and consequently creating a disordered film because of the void space generated among surfactant molecules.
  • Microemulsions may, however, be prepared without the use of cosurfactants and alcohol-free self-emulsifying microemulsion systems are known in the art.
  • the aqueous phase may typically be, but is not limited to, water, an aqueous solution of the drug, glycerol, PEG300, PEG400, polyglycerols, propylene glycols, and derivatives of ethylene glycol.
  • the oil phase may include, but is not limited to, materials such as Captex 300, Captex 355, Capmul MCM, fatty acid esters, medium chain (C8-C12) mono, di, and triglycerides, polyoxyethylated glyceryl fatty acid esters, fatty alcohols, polyglycolized glycerides, saturated polyglycolized C8-C10 glycerides, vegetable oils and silicone oil.
  • materials such as Captex 300, Captex 355, Capmul MCM, fatty acid esters, medium chain (C8-C12) mono, di, and triglycerides, polyoxyethylated glyceryl fatty acid esters, fatty alcohols, polyglycolized glycerides, saturated polyglycolized C8-C10 glycerides, vegetable oils and silicone oil.
  • Microemulsions are particularly of interest from the standpoint of drug solubilization and the enhanced abso ⁇ tion of drugs.
  • Lipid based microemulsions both o/w and w/o have been proposed to enhance the oral bioavailability of drugs, including peptides (Constantinides et al., Pharmaceutical Research, 1994, 1 1, 1385-1390; Ritschel, Meth. Find. Exp. Clin. Pharmacol, 1993, 13, 205).
  • Microemulsions afford advantages of improved drug solubilization, protection of drug from enzymatic hydrolysis, possible enhancement of drug abso ⁇ tion due to surfactant-induced alterations in membrane fluidity and permeability, ease of preparation, ease of oral administration over solid dosage forms, improved clinical potency, and decreased toxicity (Constantinides et al., Pharmaceutical Research, 1994, 11, 1385; Ho et al., J Pharm. Sci., 1996, 85, 138-143). Often microemulsions may form spontaneously when their components are brought together at ambient temperature. This may be particularly advantageous when formulating thermolabile drugs, peptides or oligonucleotides. Microemulsions have also been effective in the transdermal delivery of active components in both cosmetic and pharmaceutical applications.
  • microemulsion compositions and formulations of the present invention will facilitate the increased systemic abso ⁇ tion of oligonucleotides and nucleic acids from the gastrointestinal tract, as well as improve the local cellular uptake of oligonucleotides and nucleic acids within the gastrointestinal tract, vagina, buccal cavity and other areas of administration.
  • Microemulsions of the present invention may also contain additional components and additives such as sorbitan monostearate (Grill 3), Labrasol, and penetration enhancers to improve the properties of the formulation and to enhance the abso ⁇ tion of the oligonucleotides and nucleic acids of the present invention.
  • Penetration enhancers used in the microemulsions of the present invention may be classified as belonging to one of five broad categories - surfactants, fatty acids, bile salts, chelating agents, and non-chelating non-surfactants (Lee et al., Critical
  • Liposomes There are many organized surfactant structures besides microemulsions that have been studied and used for the formulation of drugs. These include monolayers, micelles, bilayers and vesicles. Vesicles, such as liposomes, have attracted great interest because of their specificity and the duration of action they offer from the standpoint of drug delivery. As used in the present invention, the term "liposome” means a vesicle composed of amphiphilic lipids arranged in a spherical bilayer or bilayers.
  • Liposomes are unilamellar or multilamellar vesicles which have a membrane formed from a lipophilic material and an aqueous interior. The aqueous portion contains the composition to be delivered. Cationic liposomes possess the advantage of being able to fuse to the cell wall. Noncationic liposomes, although not able to fuse as efficiently with the cell wall, are taken up by macrophages in vivo. [0069] In order to cross intact mammalian skin, lipid vesicles must pass through a series of fine pores, each with a diameter less than 50 nm, under the influence of a suitable transdermal gradient.
  • liposomes which is highly deformable and able to pass through such fine pores.
  • Further advantages of liposomes include; liposomes obtained from natural phospholipids are biocompatible and biodegradable; liposomes can inco ⁇ orate a wide range of water and lipid soluble drugs; liposomes can protect encapsulated drugs in their internal compartments from metabolism and degradation (Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, P. 245).
  • Liposomes are useful for the transfer and delivery of active ingredients to the site of action. Because the liposomal membrane is structurally similar to biological membranes, when liposomes are applied to a tissue, the liposomes start to merge with the cellular membranes. As the merging of the liposome and cell progresses, the liposomal contents are emptied into the cell where the active agent may act.
  • Liposomes present several advantages over other formulations. Such advantages include reduced side-effects related to high systemic abso ⁇ tion of the administered drug, increased accumulation of the administered drug at the desired target, and the ability to administer a wide variety of drugs, both hydrophilic and hydrophobic, into the skin.
  • liposomes to deliver agents including high-molecular weight DNA into the skin.
  • Compounds including analgesics, antibodies, hormones and high-molecular weight DNAs have been administered to the skin. The majority of applications resulted in the targeting of the upper epidermis.
  • Liposomes fall into two broad classes. Cationic liposomes are positively charged liposomes, which interact with the negatively charged DNA molecules to form a stable complex. The positively charged DNA/liposome complex binds to the negatively charged cell surface and is internalized in an endosome. Due to the acidic pH within the endosome, the liposomes are ruptured, releasing their contents into the cell cytoplasm (Wang et al., Biochem. Biophys. Res. Commun., 1987, 147, 980 - 985) [0075] Liposomes, which are pH-sensitive or negatively-charged, entrap DNA rather than complex with it. Since both the DNA and the lipid are similarly charged, repulsion rather than complex formation occurs.
  • liposomal composition includes phospholipids other than naturally-derived phosphatidylcholine.
  • Neutral liposome compositions for example, can be formed from dimyristoyl phosphatidylcholine (DMPC) or dipalmitoyl phosphatidylcholine (DPPC).
  • Anionic liposome compositions generally are formed from dimyristoyl phosphatidylglycerol, while anionic fusogenic liposomes are formed primarily from dioleoyl phosphatidylethanolamine (DOPE).
  • DOPE dioleoyl phosphatidylethanolamine
  • Another type of liposomal composition is formed from phosphatidylcholine (PC) such as, for example, soybean PC, and egg PC.
  • PC phosphatidylcholine
  • Another type is formed from mixtures of phospholipid and/or phosphatidylcholine and/or cholesterol.
  • Non-ionic liposomal systems have also been examined to determine their utility in the delivery of drugs to the skin, in particular systems comprising non-ionic surfactant and cholesterol.
  • Non-ionic liposomal formulations comprising Novasome TM I (glyceryl dilaurate/cholesterol/polyoxyethylene-10-stearyl ether) and NovasomeTM II (glyceryl distearate/ cholesterol/polyoxyethylene-10-stearyl ether) were used to deliver cyclosporin-A into the dermis of mouse skin. Results indicated that such non-ionic liposomal systems were effective in facilitating the deposition of cyclosporin-A into different layers of the skin (Hu et al. S.TP.Pharma. Sci., 1994, 4, 6, 466).
  • Liposomes also include "sterically stabilized" liposomes, a term which, as used herein, refers to liposomes comprising one or more specialized lipids that, when inco ⁇ orated into liposomes, result in enhanced circulation lifetimes relative to liposomes lacking such specialized lipids.
  • sterically stabilized liposomes are those in which part of the vesicle-forming lipid portion of the liposome (A) comprises one or more glycolipids, such as monosialoganglioside G M I , or (B) is derivatized with one or more hydrophilic polymers, such as a polyethylene glycol (PEG) moiety.
  • PEG polyethylene glycol
  • Liposomes comprising sphingomyelin. Liposomes comprising 1 ,2-sn-dimyristoylphosphatidylcholine are disclosed in WO 97/13499 (Lim et al.).
  • liposomes comprising lipids derivatized with one or more hydrophilic polymers, and methods of preparation thereof, are known in the art.
  • Sunamoto et al. (Bull. Chem. Soc. Jpn., 1980, 53, 2778) described liposomes comprising a nonionic detergent, 2C 12 15G, that contains a PEG moiety.
  • Ilium et al. (FEBS Lett., 1984, 167, 79) noted that hydrophilic coating of polystyrene particles with polymeric glycols results in significantly enhanced blood half-lives.
  • a limited number of liposomes comprising nucleic acids are known in the art.
  • WO 96/40062 to Thierry et al. discloses methods for encapsulating high molecular weight nucleic acids in liposomes.
  • U.S. Patent No. 5,264,221 to Tagawa et al. discloses protein-bonded liposomes and asserts that the contents of such liposomes may include an antisense RNA.
  • U.S. Patent No. 5,665,710 to Rahman et al. describes certain methods of encapsulating oligodeoxynucleotides in liposomes.
  • Transfersomes are yet another type of liposomes, and are highly deformable lipid aggregates which are attractive candidates for drug delivery vehicles. Transfersomes may be described as lipid droplets which are so highly deformable that they are easily able to penetrate through pores which are smaller than the droplet. Transfersomes are adaptable to the environment in which they are used, e.g. they are self-optimizing (adaptive to the shape of pores in the skin), self- repairing, frequently reach their targets without fragmenting, and often self-loading. To make transfersomes it is possible to add surface edge-activators, usually surfactants, to a standard liposomal composition. Transfersomes have been used to deliver serum albumin to the skin. The transfersome-mediated delivery of serum albumin has been shown to be as effective as subcutaneous injection of a solution containing serum albumin.
  • Surfactants find wide application in formulations such as emulsions (including microemulsions) and liposomes.
  • HLB hydrophile/lipophile balance
  • the nature of the hydrophilic group also known as the "head" provides the most useful means for categorizing the different surfactants used in formulations (Rieger, in Pharmaceutical Dosage Forms, Marcel Dekker, Inc., New York, NY, 1988, p. 285)
  • the surfactant molecule is not ionized, it is classified as a nonionic surfactant.
  • Nonionic surfactants find wide application in pharmaceutical and cosmetic products and are usable over a wide range of pH values. In general their HLB values range from 2 to about 18 depending on their structure.
  • Nonionic surfactants include nonionic esters such as ethylene glycol esters, propylene glycol esters, glyceryl esters, polyglyceryl esters, sorbitan esters, sucrose esters, and ethoxylated esters.
  • Nonionic alkanolamides and ethers such as fatty alcohol ethoxylates, propoxylated alcohols, and ethoxylated/propoxylated block polymers are also included in this class.
  • the polyoxyethylene surfactants are the most popular members of the nonionic surfactant class.
  • Anionic surfactants include carboxylates such as soaps, acyl lactylates, acyl amides of amino acids, esters of sulfuric acid such as alkyl sulfates and ethoxylated alkyl sulfates, sulfonates such as alkyl benzene sulfonates, acyl isethionates, acyl taurates and sulfosuccinates, and phosphates.
  • the most important members of the anionic surfactant class are the alkyl sulfates and the soaps.
  • Cationic surfactants include quaternary ammonium salts and ethoxylated amines. The quaternary ammonium salts are the most used members of this class.
  • the surfactant molecule has the ability to carry either a positive or negative charge, the surfactant is classified as amphoteric.
  • Amphoteric surfactants include acrylic acid derivatives, substituted alkylamides, N-alkylbetaines and phosphatides.
  • the use of surfactants in drug products, formulations and in emulsions has been reviewed (Rieger, in Pharmaceutical Dosage Forms, Marcel Dekker, Inc., New York, NY, 1988, p. 285).
  • Penetration Enhancers include acrylic acid derivatives, substituted alkylamides, N-alkylbetaines and phosphatides.
  • the present invention employs various penetration enhancers to effect the efficient delivery of nucleic acids particularly oligonucleotides, to the skin of animals.
  • Most drugs are present in solution in both ionized and nonionized forms. However, usually only lipid soluble or lipophilic drugs readily cross cell membranes. It has been discovered that even non-1 ipophilic drugs may cross cell membranes if the membrane to be crossed is treated with a penetration enhancer. In addition to aiding the diffusion of non-lipophilic drugs across cell membranes, penetration enhancers also enhance the permeability of lipophilic drugs.
  • Penetration enhancers may be classified as belonging to one of five broad categories, i.e., surfactants, fatty acids, bile salts, chelating agents, and non- chelating nonsurfactants (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991 , p.92). Each of the above mentioned classes of penetration enhancers are described below in greater detail.
  • surfactants are chemical entities which, when dissolved in an aqueous solution, reduce the surface tension of the solution or the interfacial tension between the aqueous solution and another liquid, with the result that abso ⁇ tion of oligonucleotides through the mucosa. is enhanced.
  • these penetration enhancers include, for example, sodium lauryl sulfate, polyoxyethylene- 9-lauryl ether and polyoxyethylene-20-cetyl ether) (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, p.92); and perfluorochemical emulsions, such as FC-43. Takahashi et al., J. Pharm. Pharmacol, 1988, 40, 252).
  • Various fatty acids and their derivatives which act as penetration enhancers include, for example, oleic acid, lauric acid, capric acid (n-decanoic acid), myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, dicaprate, tricaprate, monoolein (l-monooleoyl-.rac-glycerol), dilaurin, caprylic acid, arachidonic acid, glycerol 1 -monocaprate, 1 -dodecylazacycloheptan-2-one, acylcarnitines, acylcholines, C,. ⁇ o alkyl esters thereof (e.g., methyl, isopropyl and t- butyl), and mono- and di-glycerides thereof (i.e., oleate, laurate, caprate, myristate, palmitate, stearate, linoleate,
  • Bile salts [0094] The physiological role of bile includes the facilitation of dispersion and abso ⁇ tion of lipids and fat-soluble vitamins (Brunton, Chapter 38 in: Goodman & Gilman's The Pharmacological Basis of Therapeutics, 9th Ed., Hardman et al. Eds. McGraw-Hill, New York, 1996, pp. 934-935).
  • Various natural bile salts, and their synthetic derivatives act as penetration enhancers.
  • the term "bile salts" includes any of the naturally occurring components of bile as well as any of their synthetic derivatives.
  • the bile salts of the invention include, for example, cholic acid (or its pharmaceutically acceptable sodium salt, sodium cholate), dehydrocholic acid (sodium dehydrocholate), deoxycholic acid (sodium deoxycholate), glucholic acid (sodium glucholate), glycholic acid (sodium glycocholate), glycodeoxycholic acid (sodium glycodeoxycholate), taurocholic acid (sodium taurocholate), taurodeoxycholic acid (sodium taurodeoxycholate), chenodeoxycholic acid (sodium chenodeoxycholate), ursodeoxycholic acid (UDCA), sodium tauro-24,25-dihydro-fusidate (STDHF), sodium glycodihydrofusidate'and polyoxyethylene-9-lauryl ether (POE) (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, page 92; Swinyard, Chapter 39 In: Remington 's Pharmaceutical
  • Chelating agents can be defined as compounds that remove metallic ions from solution by forming complexes therewith, with the result that abso ⁇ tion of oligonucleotides through the mucosa is enhanced.
  • chelating agents have the added advantage of also serving as DNase inhibitors, as most characterized DNA nucleases require a divalent metal ion for catalysis and are thus inhibited by chelating agents (Jarrett, J. Chromatogr., 1993, 618, 315-339).
  • Chelating agents of the invention include but are not limited to disodium.
  • ethylenediaminetetraacetate citric acid
  • salicylates e.g., sodium salicylate, 5-methoxysalicylate and homovanilate
  • N-acyl derivatives of collagen e.g., laureth-9 and N-amino acyl derivatives of beta-diketones (enamines)(Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, page 92;
  • nonchelating non-surfactant penetration enhancing compounds can be defined as compounds that demonstrate insignificant activity as chelating agents or as surfactants but that nonetheless enhance abso ⁇ tion of oligonucleotides through the alimentary mucosa (Muranishi, Critical Reviews in Therapeutic Drug Carrier Systems, 1990, 7, 1-33).
  • This class of penetration enhancers include, for example, unsaturated cyclic ureas, 1 -alkyl- and 1- alkenylazacyclo-alkanone derivatives (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991 , page 92); and non-steroidal anti-inflammatory agents such as diclofenac sodium, indomethacin and phenylbutazone (Yamashita et al., J. Pharm. Pharmacol, 1987, 39, 621-626).
  • Agents that enhance uptake of oligonucleotides at the cellular level may also be added to the pharmaceutical and other compositions of the present invention.
  • cationic lipids such as lipofectin (Junichi et al, U.S. Patent No. 5,705,188), cationic glycerol derivatives, and polycationic molecules, such as polylysine (Lollo et al., PCT Application WO 97/30731), are also known to enhance the cellular uptake of oligonucleotides.
  • nucleic acids may be utilized to enhance the penetration of the administered nucleic acids, including glycols such as ethylene glycol and propylene glycol, pyrrols such as 2-pyrrol, azones, and te ⁇ enes such as limonene and menthone.
  • glycols such as ethylene glycol and propylene glycol
  • pyrrols such as 2-pyrrol
  • azones such as 2-pyrrol
  • te ⁇ enes such as limonene and menthone.
  • compositions of the present invention also inco ⁇ orate carrier compounds in the formulation.
  • carrier compound or “carrier” can refer to a nucleic acid, or analog thereof, which is inert (i.e., does not possess biological activity per se) but is recognized as a nucleic acid by in vivo processes that reduce the bioavailability of a nucleic acid having biological activity by, for example, degrading the biologically active nucleic acid or promoting its removal from circulation.
  • a nucleic acid and a carrier compound can result in a substantial reduction of the amount of nucleic acid recovered in the liver, kidney or other extracirculatory reservoirs, presumably due to competition between the carrier compound and the nucleic acid for a common receptor.
  • the recovery of a partially phosphorothioate oligonuclectide in hepatic tissue can be reduced when it is coadministered with polyinosinic acid, dextran sulfate, polycytidic acid or 4- acetamido-4'isothiocyano-stilbene-2,2'disulfonic acid (Miyao et al., Antisense Res. Dev., 1995, 5, 1 15-121 ; Takakura et al, Antisense & Nucl. Acid Drug Dev., 1996, 6, 177-183).
  • a “pharmaceutical carrier” or “excipient” is a pharmaceutically acceptable solvent, suspending agent or any other pharmacologically inert vehicle for delivering one or more nucleic acids to an animal.
  • the excipient may be liquid or solid and is selected, with the planned manner of administration in mind, so as to provide for the desired bulk, consistency, etc., when combined with a nucleic acid and the other components of a given pharmaceutical composition.
  • Typical pharmaceutical carriers include, but are not limited to, binding agents (e.g., pregelatinized maize starch, polyvinylpyrrolidone or hydroxypropyl methylcellulose, etc.); fillers (e.g., lactose and other sugars, microcrystalline cellulose, pectin, gelatin, calcium sulfate, ethyl cellulose, polyacrylates or calcium hydrogen phosphate, etc.); lubricants (e.g., magnesium stearate, talc, silica, colloidal silicon dioxide, stearic acid, metallic stearates, hydrogenated vegetable oils, corn starch, polyethylene glycols, sodium benzoate, sodium acetate, etc.); disintegrants (e.g., starch, sodium starch glycolate, etc.); and wetting agents (e.g., sodium lauryl sulphate, etc.).
  • binding agents e.g., pregelatinized maize starch, polyvinylpyrrolidone or hydroxyprop
  • compositions of the present invention Pharmaceutically acceptable organic or inorganic excipient suitable for non-parenteral administration which do not deleteriously react with nucleic acids can also be used to formulate the compositions of the present invention.
  • suitable pharmaceutically acceptable carriers include, but are not limited to, water, salt solutions, alcohols, polyethylene glycols, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethyl cellulose, polyvinylpyrrolidone and the like.
  • Formulations for topical administration of nucleic acids may include sterile and non-sterile aqueous solutions, non-aqueous solutions in common solvents such as alcohols, or solutions of the nucleic acids in liquid or solid oil bases.
  • the solutions may also contain buffers, diluents and other suitable additives.
  • Pharmaceutically acceptable organic or inorganic excipients suitable for non- parenteral administration which do not deleteriously react with nucleic acids can be used.
  • Suitable pharmaceutically acceptable excipients include, but are not limited to, water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethyl cellulose, polyvinylpyrrolidone and the like.
  • Other Components include, but are not limited to, water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethyl cellulose, polyvinylpyrrolidone and the like.
  • Other Components may additionally contain other adjunct components conventionally found in pharmaceutical compositions, at their art-established usage levels.
  • compositions may contain additional, compatible, pharmaceutically-active materials such as, for example, antipruritics, astringents, local anesthetics or anti -inflammatory agents, or may contain additional materials useful in physically formulating various dosage forms of the compositions of the present invention, such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickening agents and stabilizers.
  • additional materials useful in physically formulating various dosage forms of the compositions of the present invention, such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickening agents and stabilizers.
  • Aqueous suspensions may contain substances which increase the viscosity of the suspension including, for example, sodium carboxymethyl cellulose, sorbitol and/or dextran.
  • the suspension may also contain stabilizers.
  • compositions containing (a) one or more antisense compounds and (b) one or more other chemotherapeutic agents which function by a non-antisense mechanism.
  • chemotherapeutic agents include, but are not limited to, anticancer drugs such as daunorubicin, dactinomycin, doxorubicin, bleomycin, mitomycin, nitrogen mustard, chlorambucil, melphalan, cyclophosphamide, 6- mercaptopurine, 6-thioguanine, cytarabine (CA), 5-fluorouracil (5-FU), floxuridine (5-FUdR), methotrexate (MTX), colchicine, vincristine, vinblastine, etoposide, teniposide, cisplatin and diethylstilbestrol (DES).
  • anticancer drugs such as daunorubicin, dactinomycin, doxorubicin, bleomycin, mitomycin, nitrogen mustard, chlorambuci
  • Anti-inflammatory drugs including but not limited to nonsteroidal anti-inflammatory drugs and corticosteroids, and antiviral drugs, including but not limited to ribivirin, vidarabine, acyclovir and ganciclovir, may also be combined in compositions of the invention.
  • compositions of the invention may contain one or more antisense compounds, particularly oligonucleotides, targeted to a first nucleic acid and one or more additional antisense compounds targeted to a second nucleic acid target. Numerous examples of antisense compounds are known in the art. Two or more combined compounds may be used together or sequentially.
  • compositions and their subsequent administration is believed to be within the skill of those in the art. Dosing is dependent on severity and responsiveness of the disease state to be treated, with the course of treatment lasting from several days to several months, or until a cure is effected or a diminution of the disease state is achieved. Optimal dosing schedules can be calculated from measurements of drug accumulation in the body of the patient. Persons of ordinary skill can easily determine optimum dosages, dosing methodologies and repetition rates. Optimum dosages may vary depending on the relative potency of individual oligonucleotides, and can generally be estimated based on EC 5 os found to be effective in in vitro and in vivo animal models.
  • dosage is from 0.01 ⁇ g to 100 g per kg of body weight, and may be given once or more daily, weekly, monthly or yearly, or even once every 2 to 20 years. Persons of ordinary skill in the art can easily estimate repetition rates for dosing based on measured residence times and concentrations of the drug in bodily fluids or tissues. Following successful treatment, it may be desirable to have the patient undergo maintenance therapy to prevent the recurrence of the disease state, wherein the oligonucleotide is administered in maintenance doses, ranging from 0.01 ⁇ g to 100 g per kg of body weight, once or more daily, to once every 20 years.
  • 2'-Deoxy and 2'-methoxy beta-cyanoethyldiisopropyl phosphoramidites are available from commercial sources (e.g. Chemgenes, Needham MA or Glen Research, Inc. Sterling VA).
  • Other 2'-O-alkoxy substituted nucleoside amidites are prepared as described in U.S. Patent 5,506,351, herein inco ⁇ orated by reference.
  • the standard cycle for unmodified oligonucleotides is utilized, except the wait step after pulse delivery of tetrazole and base is increased to 360 seconds.
  • Oligonucleotides containing 5-methyl-2'-deoxycytidine (5-Me-C) nucleotides are synthesized according to published methods [Sanghvi, et. al., Nucleic Acids Research, 1993, 21 , 3197-3203] using commercially available phosphoramidites (Glen Research, Sterling VA or ChemGenes, Needham MA). 2'-Fluoro amidites 2'-Fluorodeoxyadenosine amidites
  • 2'-fluoro oligonucleotides are synthesized as described previously [Kawasaki, et. al., J. Med. Chem., 1993, 36, 831-841 ] and United States patent 5,670,633, herein inco ⁇ orated by reference. Briefly, the protected nucleoside N6- benzoyl-2'-deoxy-2'-fluoroadenosine is synthesized utilizing commercially available 9-beta-D-arabinofuranosyladenine as starting material and by modifying literature procedures whereby the 2'-alpha-fluoro atom is introduced by a S N 2- displacement of a 2'-beta-trityl group.
  • N6-benzoyl-9-beta-D- arabinofuranosyladenine is selectively protected in moderate yield as the 3 ',5'- ditetrahydropyranyl (THP) intermediate.
  • THP 3 ',5'- ditetrahydropyranyl
  • Deprotection of the THP and N6-benzoyl groups is accomplished using standard methodologies and standard methods are used to obtain the 5'-dimethoxytrityl-(DMT) and 5'-DMT-3'-phosphoramidite intermediates.
  • Synthesis of 2 '-deoxy-2 '-fluorouridine is accomplished by the modification of a literature procedure in which 2,2'anhydro-l-beta-D- arabinofuranosyluracil is treated with 70% hydrogen fluoride-pyridine. Standard procedures are used to obtain the 5'-DMT and 5'-DMT-3'-phosphoramidites. 2'-Fluorodeoxycytidine
  • 2 '-deoxy-2 '-fluorocytidine is synthesized via amination of 2'-deoxy-2'- fluorouridine, followed by selective protection to give N4-benzoyl-2 '-deoxy-2 '- fluorocytidine. Standard procedures are used to obtain the 5'-DMT and 5'-DMT- 3 'phosphoramidites.
  • 2'-O-Methoxyethyl-substituted nucleoside amidites are prepared as follows, or alternatively, as per the methods of Martin, P., Helvetica Chimica Acta, 1995, 78, 486-504.
  • the ether is decanted and the residue is dissolved in a minimum amount of methanol (ca. 400 mL).
  • the solution is poured into fresh ether (2.5 L) to yield a stiff gum.
  • the ether is decanted and the gum is dried in a vacuum oven (60°C at 1 mm Hg for 24 h) to give a solid that is crushed to a light tan powder.
  • the material is used as is for further reactions (or it can be purified further by column chromatography using a gradient of methanol in ethyl acetate (10-25%) to give a white solid.
  • a first solution is prepared by dissolving 3'-O-acetyl-2'-O- methoxyethyl-5 '-O-dimethoxytrityl-5-methyluridine (96 g, 0.144 M) in CH 3 CN (700 mL) and set aside. Triethylamine (189 mL, 1.44 M) is added to a solution of triazole (90 g, 1.3 M) in CH 3 CN (1 L), cooled to -5°C and stirred for 0.5 h using an overhead stirrer.
  • POCl 3 is added dropwise, over a 30 minute period, to the stirred solution maintained at 0-10 ° C, and the resulting mixture stirred for an additional 2 hours.
  • the first solution is added dropwise, over a 45 minute period, to the latter solution.
  • the resulting reaction mixture is stored overnight in a cold room. Salts are filtered from the reaction mixture and the solution is evaporated. The residue is dissolved in EtOAc (1 L) and the insoluble solids are removed by filtration. The filtrate is washed with 1x300 mL of NaHCO 3 and 2x300 mL of saturated NaCl, dried over sodium sulfate and evaporated. The residue is triturated with EtOAc to give the title compound.
  • N4-Benzoyl-2'-O-methoxyethyl-5'-O-dimethoxytrityl-5-methyIcytidine-3'- amidite N4-Benzoyl-2'-O-methoxyethyl-5'-O-dimethoxytrityl-5-methylcytidine (74 g, 0.10 M) is dissolved in CH 2 C1 2 (1 L) Tetrazole diisopropylamine (7.1 g) and 2-cyanoethoxy-tetra(isopropyl)phosphite (40.5 mL, 0.123 M) are added with stirring, under a nitrogen atmosphere.
  • the resulting mixture is stirred for 20 hours at room temperature (TLC showed the reaction to be 95% complete).
  • the reaction mixture is extracted with saturated NaHCO 3 (1x300 mL) and saturated NaCl (3x300 mL).
  • the aqueous washes are back-extracted with CH C1 2 (300 mL), and the extracts are combined, dried over MgSO 4 and concentrated.
  • the residue obtained is chromatographed on a 1.5 kg silica column using EtOAc/hexane (3:1) as the eluting solvent. The pure fractions were combined to give the title compound.
  • 2'-(Dimethylaminooxyethoxy) nucleoside amidites [also known in the art as 2'-O-(dimethylaminooxyethyl) nucleoside amidites] are prepared as described in the following paragraphs.
  • Adenosine, cytidine and guanosine nucleoside amidites are prepared similarly to the thymidine (5-methyluridine) except the exocyclic amines are protected with a benzoyl moiety in the case of adenosine and cytidine and with isobutyryl in the case of guanosine.
  • reaction vessel is cooled to ambient and opened.
  • TLC Rf 0.67 for desired product and Rf 0.82 for ara-T side product, ethyl acetate
  • 70%> conversion to the product In order to avoid additional side product formation, the reaction is stopped, concentrated under reduced pressure (10 to 1mm, Hg) in a warm water bath (40-100°C) with the more extreme conditions used to remove the ethylene glycol. [Alternatively, once the low boiling solvent is gone, the remaining solution can be partitioned between ethyl acetate and water. The product will be in the organic phase.] The residue is purified by column chromatography (2kg silica gel, ethyl acetate-hexanes gradient 1 :1 to 4:1).
  • reaction mixture is flushed with argon and dry THF (369.8mL, Aldrich, sure seal bottle) is added to get a clear solution.
  • Diethyl- azodicarboxylate (6.98mL, 44.36mmol) is added dropwise to the reaction mixture. The rate of addition is maintained such that resulting deep red coloration is just discharged before adding the next drop.
  • the reaction is stirred for 4 hrs. By that time TLC showed the completion of the reaction (ethylacetate:hexane, 60:40). The solvent is evaporated in vacuum.
  • Residue obtained is placed on a flash column and eluted with ethyl acetate :hexane (60:40), to get 2'-O-([2-phthalimidoxy)ethyl]-5'-t-butyldiphenylsilyl-5-methyluridine as white foam.
  • Aqueous NaHCO 3 solution (5%, lOmL) is added and extracted with ethyl acetate (2x20mL). Ethyl acetate phase is dried over anhydrous Na 2 SO 4 , evaporated to dryness.
  • Residue is dissolved in a solution of IM PPTS in MeOH (30.6mL).
  • Formaldehyde (20% w/w, 30mL, 3.37mmol) is added and the reaction mixture is stirred at room temperature for 10 minutes.
  • reaction mixture is removed from the ice bath and stirred at room temperature for 2 hrs.
  • 5% NaHCO 3 (25mL) solution is added and extracted with ethyl acetate (2x25mL).
  • Ethyl acetate layer is dried over anhydrous Na 2 SO 4 and evaporated to dryness.
  • the residue obtained is purified by flash column chromatography and eluted with 5% MeOH in CH 2 C1 2 to get 5'-O- tertbutyldiphenylsilyl-2'-O-[N,N-dimethylaminooxyethyl]-5- methyluridine as a white foam.
  • Triethylamine trihydrofluoride (3.91 mL, 24.0mmol) is dissolved in dry THF and triethylamine (1.67mL, 12mmol, dry, kept over KOH). This mixture of triethylamine-2HF is then added to 5'-O-tert-butyldiphenylsilyl-2'-O-[N,N- dimethylaminooxyethyl]-5-methyluridine (1.40g, 2.4mmol) and stirred at room temperature for 24 hrs. Reaction is monitored by TLC (5% MeOH in CH C1 2 ). Solvent is removed under vacuum and the residue placed on a flash column and eluted with 10% MeOH in CH 2 C1 2 to get 2'-O-(dimethylaminooxyethyl)-5- methyluridine.
  • reaction mixture is dissolved in anhydrous acetonitrile (8.4mL) and 2-cyanoethyl-N,N,N',N'-tetraisopropylphosphoramidite (2.12mL, 6.08mmol) is added.
  • the reaction mixture is stirred at ambient temperature for 4 hrs under inert atmosphere.
  • the progress of the reaction is monitored by TLC (hexane:ethyl acetate 1 : 1).
  • the solvent is evaporated, then the residue is dissolved in ethyl acetate (70mL) and washed with 5% aqueous NaHC0 (40mL). Ethyl acetate layer is dried over anhydrous Na SO 4 and concentrated.
  • Residue obtained is chromatographed (ethyl acetate as eluent) to get 5'-O-DMT-2'- O-(2-N,N-dimethylaminooxyethyl)-5-methyluridine-3'-[(2-cyanoethyl)-N,N- diisopropylphosphoramidite] as a foam.
  • 2'-(Aminooxyethoxy) nucleoside amidites [also known in the art as 2'- O-(aminooxyethyl) nucleoside amidites] are prepared as described in the following paragraphs. Adenosine, cytidine and thymidine nucleoside amidites are prepared similarly. N2-isobutyryl-6-O-diphenylcarbamoyl-2'-O-(2-ethylacetyl)-5'-O-(4,4'- dimethoxytrityl)guanosine-3'-[(2-cyanoethyl)-N,N- diisopropylphosphoramidite]
  • the 2'-O-aminooxyethyl guanosine analog may be obtained by selective 2'-0-alkylation of diaminopurine riboside.
  • Multigram quantities of diaminopurine riboside may be purchased from Schering AG (Berlin) to provide 2'-O-(2- ethylacetyl) diaminopurine riboside along with a minor amount of the 3'-O-isomer.
  • 2'-O-(2-ethylacetyl) diaminopurine riboside may be resolved and converted to 2'- O-(2ethylacetyl)guanosine by treatment with adenosine deaminase.
  • Standard protection procedures should afford 2'-O-(2-ethylacetyl)-5 '-O-(4,4'- dimethoxytrityl)guanosine and 2-N-isobutyryl-6-O-diphenylcarbamoyl-2 '-O-(2- ethylacetyl)-5'-O-(4,4'-dimethoxytrityl)guanosine which may be reduced to provide 2-N-isobutyryl-6-O-diphenylcarbamoyl-2'-O-(2-ethylacetyl)-5'-O-(4,4'- dimethoxytrityl)guanosine.
  • the hydroxyl group may be displaced by N- hydroxyphthalimide via a Mitsunobu reaction, and the protected nucleoside may phosphitylated as usual to yield 2-N-isobutyryl-6-O-diphenylcarbamoyl-2'-O-(2- ethylacetyl)-5'-O-(4,4'-dimethoxytrityl)guanosine-3'-[(2-cyanoethyl)-N,N- diisopropylphosphoramiditel.
  • 2'-dimethyIaminoethoxyethoxy (2'-DMAEOE) nucleoside amidites [00137] 2 '-dimethylaminoethoxyethoxy nucleoside amidites (also known in the art as 2'-O-dimethylaminoethoxyethyl, i.e., 2O-CH 2 -O-CH 2 -N(CH2) 2 , or 2'- DMAEOE nucleoside amidites) are prepared as follows. Other nucleoside amidites are prepared similarly.
  • the bomb is cooled to room temperature and opened.
  • the crude solution is concentrated and the residue partitioned between water (200 mL) and hexanes (200 mL).
  • the excess phenol is extracted into the hexane layer.
  • the aqueous layer is extracted with ethyl acetate (3x200 mL) and the combined organic layers are washed once with water, dried over anhydrous sodium sulfate and concentrated.
  • the residue is columned on silica gel using methanol/methylene chloride 1 :20 (which has 2%> triethylamine) as the eluent. As the column fractions are concentrated a colorless solid forms which is collected to give the title compound as a white solid.
  • the thiation wait step is increased to 68 sec and is followed by the capping step.
  • the oligonucleotides are purified by precipitating twice with 2.5 volumes of ethanol from a 0.5 M NaCl solution.
  • Phosphinate oligonucleotides are prepared as described in U.S. Patent
  • Alkyl phosphonate oligonucleotides are prepared as described in U.S.
  • 3 '-Deoxy-3 '-methylene phosphonate oligonucleotides are prepared as described in U.S. Patents 5,610,289 or 5,625,050, herein inco ⁇ orated by reference.
  • Phosphoramidite oligonucleotides are prepared as described in U.S.
  • Alkylphosphonothioate oligonucleotides are prepared as described in WO 94/17093 and WO 94/02499 herein inco ⁇ orated by reference.
  • 3 '-Deoxy-3 '-amino phosphoramidate oligonucleotides are prepared as described in U.S. Patent 5,476,925, herein inco ⁇ orated by reference.
  • Phosphotriester oligonucleotides are prepared as described in U.S.
  • Formacetal and thioformacetal linked oligonucleosides are prepared as described in U.S. Patents 5,264,562 and 5,264,564, herein inco ⁇ orated by reference.
  • Ethylene oxide linked oligonucleosides are prepared as described in U.S. Patent 5,223,618, herein inco ⁇ orated by reference.
  • PNAs Peptide nucleic acids
  • PNA Peptide nucleic acids
  • Chimeric oligonucleotides, oligonucleosides or mixed oligonucleotides/oligonucleosides of the invention can be of several different types. These include a first type wherein the "gap" segment of linked nucleosides is positioned between 5' and 3' "wing" segments of linked nucleosides and a second "open end” type wherein the "gap” segment is located at either the 3' or the 5' terminus of the oligomeric compound. Oligonucleotides of the first type are also known in the art as “gapmers” or gapped oligonucleotides.
  • Oligonucleotides of the second type are also known in the art as “hemimers” or “wingmers”. [2'-O-Me]— [2'-deoxy]--[2'-O-Me] Chimeric Phosphorothioate Oligonucleotides [00155] Chimeric oligonucleotides having 2'-O-alkyl phosphorothioate and 2'- deoxy phosphorothioate oligonucleotide segments are synthesized using an Applied Biosystems automated DNA synthesizer Model 380B, as above.
  • Oligonucleotides are synthesized using the automated synthesizer and 2'-deoxy-5'-dimethoxytrityl- 3'-O-phosphoramidite for the DNA portion and 5'-dimethoxytrityl-2'-O-methyl-3'- O-phosphoramidite for 5' and 3' wings.
  • the standard synthesis cycle is modified by increasing the wait step after the delivery of tetrazole and base to 600 s repeated four times for RNA and twice for 2'-O-methyl.
  • the fully protected oligonucleotide is cleaved from the support and the phosphate group is deprotected in 3:1 ammonia/ethanol at room temperature overnight then lyophilized to dryness.
  • [00156] [2'-O-(2-methoxyethyl)]-[2'-deoxy]—[-2'-O-(methoxyethyl)] chimeric phosphorothioate oligonucleotides are prepared as per the procedure above for the 2'-O-methyl chimeric oligonucleotide, with the substitution of phorothioate oligonucleotides are prepared as per the procedure abo 2'-O-(methoxyethyl) amidites for the 2'-O-methyl amidites.
  • oligonucleotides or oligonucleosides are purified by precipitation twice out of 0.5 M NaCl with 2.5 volumes ethanol. Synthesized oligonucleotides are analyzed by polyacrylamide gel electrophoresis on denaturing gels and judged to be at least 85% full length material. The relative amounts of phosphorothioate and phosphodiester linkages obtained in synthesis are periodically checked by P nuclear magnetic resonance spectroscopy, and for some studies oligonuclectides are purified by HPLC, as described by Chiang et al., J. Biol. Chem. 1991, 266, 18162- 18171.
  • Oligonucleotides are synthesized via solid phase P(III) phosphoramidite chemistry on an automated synthesizer capable of assembling 96 sequences simultaneously in a standard 96 well format.
  • Phosphodiester internucleotide linkages are afforded by oxidation with aqueous iodine.
  • Phosphorothioate internucleotide linkages are generated by sulfurization utilizing 3,H-1,2 benzodithiole-3-one 1 ,1 dioxide (Beaucage Reagent) in anhydrous acetonitrile.
  • Standard base -protected beta-cyanoethyldiisopropyl phosphoramidites can be purchased from commercial vendors (e.g.
  • Non-standard nucleosides are synthesized as per known literature or patented methods. They are utilized as base protected betacyanoethyldiisopropyl phosphoramidites.
  • Oligonucleotides are cleaved from support and deprotected with concentrated NH OH at elevated temperature (55-60°C) for 12-16 hours and the released product then dried in vacuo. The dried product is then re-suspended in sterile water to afford a master plate from which all analytical and test plate samples are then diluted utilizing robotic pipettors.
  • oligonucleotide concentration is assessed by dilution of samples and UV absorption spectroscopy.
  • the full-length integrity of the individual products is evaluated by capillary electrophoresis (CE) in either the 96 well format (Beckman P/ACETM MDQ) or, for individually prepared samples, on a commercial CE apparatus (e.g., Beckman P/ACETM 5000, ABI 270). Base and backbone composition is confirmed by mass analysis of the compounds utilizing electrospray-mass spectroscopy. All assay test plates are diluted from the master plate using single and multi-channel robotic pipettors. Plates are judged to be acceptable if at least 85% of the compounds on the plate are at least 85% full length.
  • the effect of antisense compounds on target nucleic acid expression can be tested in any of a variety of cell types provided that the target nucleic acid is present at measurable levels. This can be routinely determined using, for example, PCR or Northern blot analysis. The following 6 cell types are provided for illustrative pu ⁇ oses, but other cell types can be routinely used, provided that the target is expressed in the cell type chosen. This can be readily determined by methods routine in the art, for example Northern blot analysis, Ribonuclease protection assays, or RT-PCR.
  • T-24 cells The human transitional cell bladder carcinoma cell line T-24 is obtained from the American Type Culture Collection (ATCC) (Manassas, VA). T-24 cells are routinely cultured in complete McCoy's 5A basal media (Gibco/Life Technologies, Gaithersburg, MD) supplemented with 10% fetal calf serum (Gibco/Life Technologies, Gaithersburg, MD), penicillin 100 units per mL, and streptomycin 100 micrograms per mL (Gibco/Life Technologies, Gaithersburg, MD). Cells are routinely passaged by trypsinization and dilution when they reached 90% confluence.
  • ATCC American Type Culture Collection
  • Cells are seeded into 96-well plates (Falcon-Primaria #3872) at a density of 7000 cells/well for use in RT-PCR analysis.
  • cells may be seeded onto 100 mm or other standard tissue culture plates and treated similarly, using appropriate volumes of medium and oligonucleotide.
  • A549 cells [00166] The human lung carcinoma cell line A549 can be obtained from the American Type Culture Collection (ATCC) (Manassas, VA). A549 cells are routinely cultured in DMEM basal media (Gibco/Life Technologies, Gaithersburg, MD) supplemented with 10% fetal calf serum (Gibco/Life Technologies, Gaithersburg, MD), penicillin 100 units per mL, and streptomycin 100 micrograms per mL (Gibco/Life Technologies, Gaithersburg, MD). Cells are routinely passaged by trypsinization and dilution when they reached 90% confluence.
  • ATCC American Type Culture Collection
  • Human neonatal dermal fibroblast can be obtained from the Clonetics Co ⁇ oration (Walkersville MD). NHDFs are routinely maintained in Fibroblast Growth Medium (Clonetics Co ⁇ oration, Walkersville MD) supplemented as recommended by the supplier. Cells are maintained for up to 10 passages as recommended by the supplier.
  • HEK Human embryonic keratinocytes
  • Clonetics Corporation Walkersville MD
  • HEKs are routinely maintained in Keratinocyte Growth Medium (Clonetics Corporation, Walkersville MD) formulated as recommended by the supplier.
  • Cells are routinely maintained for up to 10 passages as recommended by the supplier.
  • MCF-7 cells [00169] The human breast carcinoma cell line MCF-7 is obtained from the American Type Culure Collection (Manassas, VA). MCF-7 cells are routinely cultured in DMEM low glucose (Gibco/Life Technologies, Gaithersburg, MD) supplemented with 10%> fetal calf serum (Gibco/Life Technologies, Gaithersburg, MD). Cells are routinely passaged by trypsinization and dilution when they reached 90% confluence. Cells are seeded into 96-well plates (Falcon-Primaria #3872) at a density of 7000 cells/well for use in RT-PCR analysis.
  • cells may be seeded onto 100 mm or other standard tissue culture plates and treated similarly, using appropriate volumes of medium and oligonucleotide.
  • the mouse lung epithelial cell line LA4 is obtained from the American Type Culure Collection (Manassas, VA). LA4 cells are routinely cultured in F12K medium (Gibco/Life Technologies, Gaithersburg, MD) supplemented with 15% fetal calf serum (Gibco/Life Technologies, Gaithersburg, MD). Cells are routinely passaged by trypsinization and dilution when they reached 90% confluence. Cells are seeded into 96-well plates (Falcon-Primaria #3872) at a density of 3000-6000 cells/ well for use in RT-PCR analysis.
  • cells may be seeded onto 100 mm or other standard tissue culture plates and treated similarly, using appropriate volumes of medium and oligonucleotide.
  • Treatment with antisense compounds [00174] When cells reached 80% confluency, they are treated with oligonucleotide. For cells grown in 96-well plates, wells are washed once with 200 ⁇ L OPTl-MEM ,m -] reduced-serum medium (Gibco BRL) and then treated with 130 ⁇ L of OPTI-MEMTMTM-l containing 3.75 ⁇ g/mL LIPOFECT1NTM (Gibco BRL) and the desired concentration of oligonucleotide. After 4-7 hours of treatment, the medium is replaced with fresh medium. Cells are harvested 16-24 hours after oligonucleotide treatment.
  • the concentration of oligonucleotide used varies from cell line to cell line. To determine the optimal oligonucleotide concentration for a particular cell line, the cells are treated with a positive control oligonucleotide at a range of concentrations. If 80% inhibition is not achieved, the lowest concentration of positive control oligonucleotide that results in 60% inhibition of H-ras or c-raf mRNA is then utilized as the oligonucleotide screening concentration in subsequent experiments for that cell line. If 60% inhibition is not achieved, that particular cell line is deemed as unsuitable for oligonucleotide transfection experiments.
  • Antisense modulation of GR expression can be assayed in a variety of ways known in the art.
  • GR mRNA levels can be quantitated by, e.g., Northern blot analysis, competitive polymerase chain reaction (PCR), or real-time PCR (RT-PCR). Real-time quantitative PCR is presently preferred.
  • RNA analysis can be performed on total cellular RNA or poly(A)+ mRNA. Methods of RNA isolation are taught in, for example, Ausubel, F.M. et al., Current Protocols in Molecular Biology, Volume 1, pp. 4.1.1-4.2.9 and 4.5.1-4.5.3, John Wiley & Sons, Inc., 1993.
  • mRNA isolated from untreated cells is serially diluted. Each dilution is amplified in the presence of primer-probe sets specific for GAPDH only, target gene only ("single-plexing"), or both (multiplexing). Following PCR amplification, standard curves of GAPDH and target mRNA signal as a function of dilution are generated from both the single- plexed and multiplexed samples. If both the slope and correlation coefficient of the GAPDH and target signals generated from the multiplexed samples fall within 10% of their corresponding values generated from the single-plexed samples, the primer- probe set specific for that target is deemed as multiplexable. Other methods of PCR are also known in the art.
  • Protein levels of GR can be quantitated in a variety of ways well known in the art, such as immunoprecipitation, Western blot analysis (immunoblotting), ELISA or fluorescence-activated cell sorting (FACS).
  • Antibodies directed to GR can be identified and obtained from a variety of sources, such as the MSRS catalog of antibodies (Aerie Corporation, Birmingham, MI), or can be prepared via conventional antibody generation methods. Methods for preparation of polyclonal antisera are taught in, for example, Ausubel, F.M. et al., Current Protocols in Molecular Biology, Volume 2, pp. 11.12.1-11.12.9, John Wiley & Sons, Inc., 1997. Preparation of monoclonal antibodies is taught in, for example, Ausubel, F.M. et al., Current Protocols in Molecular Biology, Volume 2, pp. 11.4.1 -11.11.5, John Wiley Sons, Inc., 1997.
  • Immunoprecipitation methods are standard in the art and can be found at, for example, Ausubel, F.M. et al., Current Protocols in Molecular Biology, Volume 2, pp. 10.16.1 10.16.11, John Wiley & Sons, Inc., 1998.
  • Western blot (immunoblot) analysis is standard in the art and can be found at, for example, Ausubel, F.M. et al., Current Protocols in Molecular Biology, Volume 2, pp. 10.8.1-10.8.21 , John Wiley Sons, Inc., 1997.
  • Enzyme-linked immunosorbent assays ELISA are standard in the art and can be found at, for example, Ausubel, F.M. et al., Current Protocols in Molecular Biology, Volume 2, pp. 1 1.2.1-11.2.22, John Wiley & Sons, Inc., 1991.
  • Poly(A)+ mRNA isolation [00179] Poly(A)+ mRNA is isolated according to Miura et al., Clin. Chem., 1996, 42, 1758-1764. Other methods for poly(A)+ mRNA isolation are taught in, for example, Ausubel, F.M. et al., Current Protocols in Molecular Biology, Volume 1 , pp. 4.5.1-4.5.3, John Wiley & Sons, Inc., 1993. Briefly, for cells grown on 96- well plates, growth medium is removed from the cells and each well is washed with 200 ⁇ L cold PBS.
  • lysis buffer (10 mM Tris-HCl, pH 7.6, 1 mM EDTA, 0.5 M NaCl, 0.5%) NP-40, 20 mM vanadyl-ribonucleoside complex
  • NP-40 20 mM vanadyl-ribonucleoside complex
  • 55 ⁇ L of lysate is transferred to Oligo d(T) coated 96-well plates (AGCT Inc., Irvine CA). Plates are incubated for 60 minutes at room temperature, washed 3 times with 200 ⁇ L of wash buffer (10 mM Tris-HCl pH 7.6, 1 mM EDTA, 0.3 M NaCl).
  • the plate is blotted on paper towels to remove excess wash buffer and then air-dried for 5 minutes.
  • 60 pL of elution buffer (5 mM Tris- HCl pH 7.6), preheated to 70°C is added to each well, the plate is incubated on a 90°C hot plate for 5 minutes, and the eluate is then transferred to a fresh 96-well plate.
  • Total mRNA is isolated using an RNEASY 96TM kit and buffers purchased from Qiagen Inc. (Valencia CA) following the manufacturer's recommended procedures. Briefly, for cells grown on 96-well plates, growth medium is removed from the cells and each well is washed with 200 ⁇ L cold PBS. 100 ⁇ L Buffer RLT is added to each well and the plate vigorously agitated for 20 seconds. 100 ⁇ L of 70%> ethanol is then added to each well and the contents mixed by pipetting three times up and down. The samples are then transferred to the RNEASY 96TM well plate attached to a QIA VAC TM manifold fitted with a waste collection tray and attached to a vacuum source. Vacuum is applied for 15 seconds.
  • Buffer RW1 1 mL of Buffer RW1 is added to each well of the RNEASY 96TM plate and the vacuum again applied for 15 seconds.
  • 1 mL of Buffer RPE is then added to each well of the RNEASY 96 plate and the vacuum applied for a period of 15 seconds.
  • the Buffer RPE wash is then repeated and the vacuum is applied for an additional 10 minutes.
  • the plate is then removed from the QIAVACTM manifold and blotted dry on paper towels.
  • the plate is then re-attached to the QIAVAC TM manifold fitted with a collection tube rack containing 1.2 mL collection tubes. RNA is then eluted by pipetting 60 ⁇ L water into each well, incubating I minute, and then applying the vacuum for 30 seconds.
  • the elution step is repeated with an additional 60 ⁇ L water.
  • the repetitive pipetting and elution steps may be automated using a QIAGEN Bio-Robot 9604 (Qiagen, Inc., Valencia CA). Essentially, after lysing of the cells on the culture plate, the plate is transferred to the robot deck where the pipetting, DNase treatment and elution steps are carried out.
  • Quantitation of GR mRNA levels is determined by real-time quantitative PCR using the ABI PRISM TM 7700 Sequence Detection System (PE-Applied Biosystems, Foster City, CA) according to manufacturer's instructions. This is a closed-tube, non-gel-based, fluorescence detection system which allows high- throughput quantitation of polymerase chain reaction (PCR) products in real-time. As opposed to standard PCR, in which amplification products are quantitated after the PCR is completed, products in real-time quantitative PCR are quantitated as they accumulate.
  • ABI PRISM TM 7700 Sequence Detection System PE-Applied Biosystems, Foster City, CA
  • reporter dye e.g., JOE, FAM, or VIC, obtained from either Operon Technologies Inc., Alameda, CA or PE- Applied Biosystems, Foster City, CA
  • a quencher dye e.g., TAMRA, obtained from either Operon Technologies Inc., Alameda, CA or PE-Applied Biosystems, Foster City, CA
  • annealing of the probe to the target sequence creates a substrate that can be cleaved by the 5'- exonuclease activity of Taq polymerase.
  • cleavage of the probe by Taq polymerase releases the reporter dye from the remainder of the probe (and hence from the quencher moiety) and a sequence-specific fluorescent signal is generated.
  • additional reporter dye molecules are cleaved from their respective probes, and the fluorescence intensity is monitored at regular intervals by laser optics built into the
  • ABI PRISM 7700 Sequence Detection System In each assay, a series of parallel reactions containing serial dilutions of mRNA from untreated control samples generates a standard curve that is used to quantitate the percent inhibition after antisense oligonucleotide treatment of test samples.
  • PCR reagents can be obtained from PE-Applied Biosystems, Foster City, CA. RT-PCR reactions are carried out by adding 25 ⁇ L PCR cocktail (lx
  • TAQMAN TM buffer A 5.5 MM MgCl 2 , 300 ⁇ M each of dATP, dCTP and dGTP, 600 ⁇ M of dUTP, 100 nM each of forward primer, reverse primer, and probe, 20
  • RNAse inhibitor 1.25 Units AMPLITAQ GOLD , and 12.5 Units MuLV reverse transcriptase
  • the RT reaction is carried out by incubation for 30 minutes at 48°C.
  • 40 cycles of a two- step PCR protocol are carried out: 95°C for 15 seconds (denaturation) followed by 60°C for 1.5 minutes (annealing/extension).
  • Probes and primers to human GR were designed to hybridize to a human GR sequence shown in Figure 1 , using published sequence information (GenBank accession number NM_000176, inco ⁇ orated herein).
  • the PCR primers were: forward primer: AGGTTGTGCAAATTAACAGTCCTAACT SEQ ID NO : 4774 reverse primer: TAGTCTTTTGCAACCATCATCCA SEQ ID NO : 4775 and the PCR probe is: FA -AGCACCTAGTCCAGTGACCTGCTGGGTAAA-TAMPA SEQ ID
  • Probes and primers to mouse GR were designed to hybridize to a mouse GR sequence shown in Figure 2, using published sequence information (GenBank NM_008173 inco ⁇ orated herein). For mouse GR the PCR primers were:
  • forward primer CGGGACCACCTCCCAAAC SEQ ID NO : 4780
  • reverse primer GCACCCCATAATGGCATACC SEQ ID NO : 4781
  • the PCR probe is: FAM-TGCCTGGTGTGCTCCGATGAAGC- TAMPA SEQ ID NO : 4782 where FAM
  • PCR primers were: forward primer: AGAGAAATTTGAGGATGAGAACTTCAT SEQ ID NO : 4783 reverse primer: TTGTGTTTGGTCCAGCATTTG SEQ ID NO : 4784 and the PCR probe is: JOE- AAGCATACAGGTCCTGGCATCTTGTCCAT- TAMRA SEQ ID NO : 4785 where JOE (PE-Applied Biosystems, Foster City, CA) is the fluorescent reporter dye) and TAMRA (PEApplied Biosystems, Foster City, CA) is the quencher dye.
  • oligonucleotides are designed to target different regions of the human GR RNA, using published sequences (GenBank accession number NM_000176.1). The oligonucleotides are shown in Table 1. "Target site” indicates the first (5 '-most) nucleotide number on the particular target sequence to which the oligonucleotide binds. The indicated parameters for each oligo were predicted using RNAstructure 3.7 by David H.
  • RNA folding algorithm is based on the following research:D.H. Mathews, et al., Journal of Molecular Biology, 288, 91 1 -940, (1999). OligoWalk is based on the following research: D.H. Mathews, et al., RNA, 5, 1458-1469 (1999).
  • the Dynalign algorithm is based on the following research: D.H.
  • the oligos are sorted by the predicted free energy of the oligo interupting the predicted secondary structure of the messenger RNA.
  • All compounds in Table 1 are chimeric oligonucleotides ("gapmers") 20 nucleotides in length, composed of a central "gap" region consisting often 2'deoxynucleotides, which is flanked on both sides (5' and 3' directions) by four-nucleotide "wings".
  • the wings are composed of 2 '-methoxy ethyl (2'-MOE) nucleotides.
  • Cytidine residues in the 2'-MOE wings are 5-methylcytidines. All cytidine residues are 5-methylcytidines.
  • Target total duplex Tm of target molecular molecu Site oligo binding formation Duplex structure oligo oligo
  • Target total duplex Tm of target molecular imolecul Site oligo binding formation
  • Duplex structure oligo oligo
  • CTCCACCCCAGAGCAAATGC 1945 SEQ ID NO: 63 -30.8 -40.5 91.4 -9.7 0 -4.5
  • AAGACATTTTCGATAGCGGC 1559 SEQ ID NO: 76 -30.4 -33.6 82.5 -3.2 0 -4.5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol kcal/mol
  • Target total duplex Tm of target molecular molecul Site oligo binding formation
  • Duplex structure oligo oligo
  • Target total duplex Tm of target molecular imolecul Site oligo binding formation
  • Duplex structure oligo oligo
  • Target total duplex Tm of target molecular imolecul Site oligo binding formation
  • Duplex structure oligo oligo
  • GCGTAGTCATGATCCTCCAA 1800 SEQ ID NO: 216 -27.7 -37.2 86.3 -8.8 0 -9.1
  • GTCTTCGCTGCTTGGAGTCT 294 SEQ ID NO -.219 -27.6 -38.9 89.1 -8.5 -2.8 -10.4
  • ATGACGACTCAACTGCTTCT 2639 SEQ ID NO: 221 -27.6 -35.5 84.4 -7.9 0 -2.6
  • GGTTTAGGTGCCATCCTTCT 2756 SEQ ID NO: 222 -27.6 -37.3 88.2 -6.8 -2.9 -11.8
  • GCCACCGTTGGTGCCAGTCT 650 SEQ ID NO: 225 -27.5 -43 95.2 -12.1 -3.2 -14.2
  • TTGGGCACTGGTGGTTTAGG 2768 SEQ ID NO: 229 -27.5 -39 91.7 -9.4 -0.9 -12.3
  • GATCTCCACCCCAGAGCAAA 1948 SEQ ID NO:233 -27.4 -39.8 90 -11.9 -0.1 -4.2
  • AGAATACTGGAGATTTGAGT 3194 SEQ ID NO: 238 -27.3 -32 79.7 -4.7 0 -6.6
  • GCCTCAGCAACCTTCACTGC 4301 SEQ ID NO: 241 -27.3 -40.5 91.6 -12.4 -0.6 -3.8
  • GCAGCCGAGATAAACAACTT 54 SEQ ID NO: 242 -27.2 -34.1 83.9 -6.9 0 -3.2
  • TTCTGTTTTCACTTGGGGCA 950 SEQ ID NO: 244 -27.2 -35.5 86.3 -7.8 -0.1 -6.4
  • TGTCATCTCCAGATCCTTGG 1233 SEQ ID NO: 246 -27.2 -37.8 86.5 -9.5 -1 -6.2
  • GTGAAGACGCAGAAACCTTC 249 SEQ ID NO: 249 -27.1 -35.3 84.7 -7 -1.1 -4.8
  • GAGGAGCTGGATGGAGGAGA 1342 SEQ ID NO: 251 -27.1 -42.7 92.3 -15.6 0 -5.3
  • TCTCCACTGAAGCAGATATA 3250 SEQ ID NO: 253 -27.1 -34.8 83.5 -7.2 -0.1 -7.2
  • TAGGTACAGACAGGGCCTCT 3386 SEQ ID NO: 254 -27.1 -41.2 93.5 -12.2 -1.9 -10.7
  • AACTGTTTGGTTTCTGAGAC 4242 SEQ ID NO: 286 -26.8 -32.3 81.2 -4.9 0 -8.5
  • GGACTAGGTGCTCTATACCA 3303 SEQ ID NO: 292 -26.7 -38.2 89.4 -8.6 -2.9 -11.8
  • AATTGGTGACAGATGGGAAT 3630 SEQ ID NO: 294 -26.7 -33.9 82.1 -4.1 -3.1 -6.2
  • GCCACCTTGAATAGAAATCA 3904 SEQ ID NO: 295 -26.7 -32.9 81.5 -6.2 0 -1.3
  • CTGTTTGGTTTCTGAGACCA 4240 SEQ ID NO: 296 -26.7 -34.6 84.3 -2.7 -3.4 -18.5
  • CAGAGTTCGATGAAATCTTC 973 SEQ ID NO: 300 -26.6 -31.2 77.6 -3.6 -0.3 -9.7
  • TCTTGACAATGGCTTTTCCT 2223 SEQ ID NO: 306 -26.6 -33.5 82.6 -6.9 0 -3.8
  • TTTTGACAAGAATACTGGAG 3202 SEQ ID NO: 308 -26.6 -30.3 78.2 -3.7 0 -5.5
  • CAGGTCACTGGACTAGGTGC 3312 SEQ ID NO: 309 -26.6 -40.9 92.2 -11.3 -3 -10.7
  • Target total duplex Tm of target molecular imolecu Site oligo binding formation Duplex structure oligo oligo

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Abstract

The present invention provides compositions and methods for modulating the expression of mammalian glucocorticoid receptor. In particular, this invention relates to antisense compounds, particularly oligonucleotides, specifically hybridizable with nucleic acids encoding human glucocorticoid receptor.

Description

ANTISENSE MODULATION OF GLUCOCORTICOID RECEPTOR
EXPRESSION
The present application claimes priority under Title 35, United States Code, §119 to United States Provisional application Serial No. 60/381 ,857, filed May 20, 2002, which is incorporated by reference in entirety as if written herein.
FIELD OF THE INVENTION
[001] The present invention provides compositions and methods for modulating the expression of mammalian glucocorticoid receptor. In particular, this invention relates to antisense compounds, particularly oligonucleotides, specifically hybridizable with nucleic acids encoding human glucocorticoid receptor. Such oligonucleotides may modulate the expression of glucocorticoid receptor.
BACKGROUND OF THE INVENTION
[002] Non insulin-dependent diabetes mellitus (type II diabetes), a debilitating disease characterized by abnormal elevation of blood glucose levels, is driven by three factors: increased hepatic glucose production, inadequate clearance of glucose via insulin mediated pathways, and decreased uptake of circulating glucose by tissues. (Diabetes Review 5(3), 177-269, (1997)). Administration of agents which decrease hepatic glucose production are a fundamental approach to controlling blood glucose levels (Am. J. Physiol. 257, E35-E42 (1989). J. Clin. Endocrinol Metab. 11, 1 180-1183 (1994) and J Clin. Invest., 92, 2283-2290 (1993)).
[003] Glucocorticoids have been shown to have major influences on glucose production. Glucocorticoid excess aggravates established diabetes and glucocorticoid deficiency reduces blood glucose and improves glucose control in diabetic mice. (Diabetes Review, 1(3), 301-308, (1993). Diabetologia, 9, 376-379 (1973). Horm. Metab. Res., 9, 152-156 (1977)). The underlying mechanism responsible for these effects is believed to be glucocorticoid-induced upregulation of hepatic enzymes required for gluconeogenesis. (Recent Prog. Horm. Res., 26, 41 1-457 (1970). Physiol. Rev., 64, 170-259 (1984). [004] The glucocorticoids are lipid soluble hormones synthesized in the adrenal cortex. They readily pass through cell membranes, enter the cytoplasm of target tissues, and bind to glucocorticoid receptors in the cytoplasm by complexation with heat shock proteins. Upon binding of the hormone to its receptor, the receptor undergoes a conformational change which results in dissociation of heat shock proteins and translocation of the ligand-bound glucocorticoid receptor into the nucleus where it can either initiate or repress specific gene transcription. Transcriptional activation occurs when the ligand bound receptor complex homodimerizes, and the homodimeric receptor ligand complex binds to chromosomal DNA at sequence-specific sites in the promoter region of regulated genes. (Cell, 56, 335-344 (1989). Annu. Rev. Genet., 19, 209-215 (1989)). Among the genes which glucocorticoids up-regulate are those which play key roles in gluconeogenesis and glycogenolysis, particularly PEPCK and glucose-6- phosphatase. (Advanced Enzymology., Meister, Ed. New York, John Wiley and Sons, Inc., 203-281 (1994). and Diabetes 45, 1563-1571(1996)).
[005] Phosphoenolpyruvatecarboxy kinase (PEPCK) catalyzes the conversion of oxaloacetate to phosphoenolpyruvate and glucose-6- phosphatase catalyzes the conversion of glucose-6-phosphate into glucose, both of which are required for the synthesis of glucose from oxaloacetate in the liver. Recently, it has been shown that Aventis®(mifepristone), a potent glucocorticoid receptor antagonist, reduces mRNA levels of PEPCK and glucose-6-phosphate in the liver and causes a 50% reduction of plasma glucose levels in obese diabetic db/db transgenic mice. (J Biol. Chem. 272(50), 31475-31481 (1997)). While steroid-based glucocorticoid receptor antagonists have been useful in demonstrating efficacy for in vivo glucose lowering effects, the utility of such agents is limited due to side effects resulting from potent cross-reactivity with other steroid receptors, in particular progesterone receptor (PR) and mineralocorticoid receptor (MR).
[006] Because agents which function as glucocorticoid antagonists represent a novel approach to controlling type II diabetes, agents which antagonize the glucocorticoid receptor have been the subject of active current research for their clinical potential. Reference is made to U.S. Pat. No. 5,929,058, which discloses a method for treating type II diabetes comprising administering a combination of nonselective steroidal agents exhibiting mineralcorticoid receptor agonist activity and glucocorticoid receptor antagonist activity.
1007] Thus, it would be an important contribution to the art to provide non- steroidal, glucocorticoid-selective agents which antagonize the glucocorticoid receptor. These compounds would be particularly useful for treating type II diabetes and the symptoms thereof, such as hyperglycemia, inadequate glucose clearance, obesity, hyperinsulinemia, hypertriglyceridemia, and high circulating glucocorticoid levels. [008] Antisense technology is emerging as an effective means for reducing the expression of specific gene products and may therefore prove to be uniquely useful in a number of therapeutic, diagnostic, and research applications for the modulation of GR expression. Systemically administered antisense has been shown to accumulate and have its effect predominately in liver and to a lessor extent in fat (R. S. Geary, R. Z. Yu, and A. A. Levin, "Pharmacokinetics of phosphorothioate antisense oligodeoxynucleotides," Curr.Opin.Investig.Drugs Volume 2, Issue 4, pp. 562-573). This relative tissue specificity would be useful in the treatment of metabolic disorders, while sparing other roles of glucocorticoid receptor, such as that of immune function.
DETAILED DESCRIPTION OF THE INVENTION
[009] Figure 1 shows the cDNA sequence of human glucocorticoid receptor and the encoded protein sequence.
[0010] Figure 2 shows the cDNA sequence of murine glucocorticoid receptor and the encoded protein sequence.
SUMMARY OF THE INVENTION
[0011] The present invention is directed to antisense compounds, particularly oligonucleotides, which are targeted to a nucleic acid encoding glucocorticoid receptor, and which modulate the expression of mammalian glucocorticoid receptor (GR). Pharmaceutical and other compositions comprising the antisense compounds of the invention are also provided. Further provided are methods of modulating the expression of glucocorticoid receptor in cells or tissues comprising contacting said cells or tissues with one or more of the antisense compounds or compositions of the invention. Further provided are methods of treating an animal, particularly a human, suspected of having or being prone to a disease or condition which may be treatable by modulation of glucocorticoid receptor activity by administering a therapeutically or prophylactically effective amount of one or more of the antisense compounds or compositions of the invention.
DETAILED DESCRIPTION OF THE INVENTION
[0012] The present invention employs oligomeric antisense compounds, particularly oligonucleotides, for use in modulating the function of nucleic acid molecules encoding glucocorticoid receptor, ultimately modulating the amount of glucocorticoid receptor produced. This is accomplished by providing antisense compounds, which specifically hybridize with one or more nucleic acids encoding glucocorticoid receptor. As used herein, the terms "target nucleic acid" and "nucleic acid encoding GR" encompass DNA encoding GR, RNA (including pre-mRNA and mRNA) transcribed from such DNA, and also cDNA derived from such RNA. The specific hybridization of an oligomeric compound with its target nucleic acid interferes with the normal function of the nucleic acid. This modulation of function of a target nucleic acid by compounds, which specifically hybridize to it, is generally referred to as "antisense". The functions of DNA to be interfered with include replication and transcription. The functions of RNA to be interfered with include all vital functions such as, for example, translocation of the RNA to the site of protein translation, translation of protein from the RNA, splicing of the RNA to yield one or more mRNA species, and catalytic activity which may be engaged in or facilitated by the RNA. The overall effect of such interference with target nucleic acid function is modulation of the expression of GR. In the context of the present invention, "modulation" means either an increase (stimulation) or a decrease (inhibition) in the expression of a gene. In the context of the present invention, inhibition is the preferred form of modulation, of gene expression and mRNA is a preferred target.
[0013] It is preferred to target specific nucleic acids for antisense. "Targeting" an antisense compound to a particular nucleic acid, in the context of this invention, is a multistep process. The process usually begins with the identification of a nucleic acid sequence whose function is to be modulated. This may be, for example, a cellular gene (or mRNA transcribed from the gene) whose expression is associated with a particular disorder or disease state, or a nucleic acid molecule from an infectious agent. In the present invention, the target is a nucleic acid molecule encoding GR. The targeting process also includes determination of a site or sites within this gene for the antisense interaction to occur such that the desired effect, e.g., detection or modulation of expression of the protein, will result. Within the context of the present invention, a preferred intragenic site is the region encompassing the translation initiation or termination codon of the open reading frame (ORF) of the gene. Since, as is known in the art, the translation initiation codon is typically 5'-AUG (in transcribed mRNA molecules; 5'-ATG in the corresponding DNA molecule), the translation initiation codon is also referred to as the "AUG codon," the "start codon" or the "AUG start codon". A minority of genes have a translation initiation codon having the RNA sequence 5'-GUG, 5'-UUG or 5'-CUG, and 5'-AUA, 5'-ACG and 5'-CUG have been shown to function in vivo. Thus, the terms "translation initiation codon" and "start codon" can encompass many codon sequences, even though the initiator amino acid in each instance is typically methionine (in eukaryotes) or formyl methionine (in prokaryotes). It is also known in the art that eukaryotic and prokaryotic genes may have two or more alternative start codons, any one of which may be preferentially utilized for translation initiation in a particular cell type or tissue, or under a particular set of conditions. In the context of the invention, "start codon" and "translation initiation codon" refer to the codon or codons that are used in vivo to initiate translation of an mRNA molecule transcribed from a gene encoding GR, regardless of the sequence(s) of such codons.
[0014] It is also known in the art that a translation termination codon (or "stop codon") of a gene may have one of three sequences, i.e. 5'-UAA, 5'-UAG and 5'- UGA (the corresponding DNA sequences are 5'-TAA, 5'-TAG and 5'-TGA, respectively). The terms "start codon region" and "translation initiation codon region "refer to a portion of such an mRNA or gene that encompasses from about 25 to about 50 contiguous nucleotides in either direction (i.e., 5' or 3') from a translation initiation codon. Similarly, the terms "stop codon region" and "translation termination codon region "refer to a portion of such an mRNA or gene that encompasses from about 25 to about 50 contiguous nucleotides in either direction (i.e., 5' or 3') from a translation termination codon. [0015] The open reading frame (ORF) or "coding region," which is known in the art to refer to the region between the translation initiation codon and the translation termination codon, is also a region which may be targeted effectively. Other target regions include the 5' untranslated region (5 'UTR), known in the art to refer to the portion of an mRNA in the 5' direction from the translation initiation codon, and thus including nucleotides between the 5' cap site and the translation initiation codon of an mRNA or corresponding nucleotides on the gene, and the 3' untranslated region (3 'UTR), known in the art to refer to the portion of an mRNA in the 3' direction from the translation termination codon, and thus including nucleotides between the translation termination codon and 3' end of an mRNA or corresponding nucleotides on the gene. The 5' cap of an mRNA comprises an N7- methylated guanosine residue joined to the 5 '-most residue of the mRNA via a 5 '-5' triphosphate linkage. The 5' cap region of an mRNA is considered to include the 5' cap structure itself as well as the first 50 nucleotides adjacent to the cap. The 5' cap region may also be a preferred target region.
[0016] Although some eukaryotic mRNA transcripts are directly translated, many contain one or more regions, known as "introns," which are excised from a transcript before it is translated. The remaining (and therefore translated) regions are known as "exons" and are spliced together to form a continuous mRNA sequence. mRNA splice sites, i.e., intron-exon junctions, may also be preferred target regions, and are particularly useful in situations where aberrant splicing is implicated in disease, or where an overproduction of a particular mRNA splice product is implicated in disease. Aberrant fusion junctions due to rearrangements or deletions are also preferred targets. It has also been found that introns can also be effective, and therefore preferred, target regions for antisense compounds targeted, for example, to DNA or pre-mRNA. [0017] Once one or more target sites have been identified, oligonucleotides are chosen which are sufficiently complementary to the target, i.e., hybridize sufficiently well and with sufficient specificity, to give the desired effect. |0018] In the context of this invention, "hybridization" means hydrogen bonding, which may be Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding, between complementary nucleoside or nucleotide bases. For example, adenine and thymine are complementary nucleobases, which pair through the formation of hydrogen bonds. "Complementary," as used herein, refers to the capacity for precise pairing between two nucleotides. For example, if a nucleotide at a certain position of an oligonucleotide is capable of hydrogen bonding with a nucleotide at the same position of a DNA or RNA molecule, then the oligonucleotide and the DNA or RNA are considered to be complementary to each other at that position. The oligonucleotide and the DNA or RNA are complementary to each other when a sufficient number of corresponding positions in each molecule are occupied by nucleotides which can hydrogen bond with each other. Thus, "specifically hybridizable" and "complementary" are terms which are used to indicate a sufficient degree of complementarity or precise pairing such that stable and specific binding occurs between the oligonucleotide and the DNA or RNA target. It is understood in the art that the sequence of an antisense compound need not be 100% complementary to that of its target nucleic acid to be specifically hybridizable. An antisense compound is specifically hybridizable when binding of the compound to the target DNA or RNA molecule interferes with the normal function of the target DNA or RNA to cause a loss of utility, and there is a sufficient degree of complementarity to avoid non-specific binding of the antisense compound to non-target sequences under conditions in which specific binding is desired, i.e., under physiological conditions in the case of in vivo assays or therapeutic treatment, and in the case of in vitro assays, under conditions in which the assays are performed. |0019] Antisense compounds are commonly used as research reagents and diagnostics. For example, antisense oligonucleotides, which are able to inhibit gene expression with exquisite specificity, are often used by those of ordinary skill to elucidate the function of particular genes. Antisense compounds are also used, for example, to distinguish between functions of various members of a biological pathway. Antisense modulation has, therefore, been harnessed for research use. [0020] The specificity and sensitivity of antisense is also harnessed by those of skill in the art for therapeutic uses. Antisense oligonucleotides have been employed as therapeutic moieties in the treatment of disease states in animals and man.
Antisense oligonucleotides have been safely and effectively administered to humans and numerous clinical trials are presently underway. It is thus established that oligonucleotides can be useful therapeutic modalities that can be configured to be useful in treatment regimes for treatment of cells, tissues and animals, especially humans. In the context of this invention, the term "oligonucleotide" refers to an oligomer or polymer of ribonucleic acid (RNA) or deoxyribonucleic acid (DNA) or mimetics thereof. This term includes oligonucleotides composed of naturally occurring nucleobases, sugars and covalent internucleoside (backbone) linkages as well as oligonucleotides having non-naturally occurring portions which function similarly. Such modified or substituted oligonucleotides are often preferred over native forms because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for nucleic acid target and increased stability in the presence of nucleases. [0021] While antisense oligonucleotides are a preferred form of antisense compound, the present invention comprehends other oligomeric antisense compounds, including but not limited to oligonuclectide mimetics such as are described below. The antisense compounds in accordance with this invention preferably comprise 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, or 50 nucleobases. The antisense compounds in accordance with this invention preferably comprise from about 8 to about 30 nucleobases (i.e. from about 8 to about 30 linked nucleo sides). Particularly preferred antisense compounds are antisense oligonuclectides, even more preferably those comprising from about 12 to about 25 nucleobases. The antisense compounds in accordance with this invention preferably comprise a nucleic acid sequence selected from the group consisiting of SEQ ID NO: 1 -SEQ ID NO:4769, and a fragment of at least eight contiguous nucleotides of SEQ ID NO: 1 -SEQ ID NO:4769. As is known in the art, a nucleoside is a base-sugar combination. The base portion of the nucleoside is normally a heterocyclic base. The two most common classes of such heterocyclic bases are the purines and the pyrimidines. Nucleotides are nucleosides that further include a phosphate group covalently linked to the sugar portion of the nucleoside. For those nucleosides that include a pentofuranosyl sugar, the phosphate group can be linked to either the 2', 3' or 5' hydroxyl moiety of the sugar. In forming oligonucleotides, the phosphate groups covalently link adjacent nucleosides to one another to form a linear polymeric compound. In turn the respective ends of this linear polymeric structure can be further joined to form a circular structure, however, open linear structures are generally preferred. Within the oligonucleotide structure, the phosphate groups are commonly referred to as forming the internucleoside backbone of the oligonucleotide. The normal I linkage or backbone of RNA and DNA is a 3' to 5' phosphodiester linkage. [0022] Specific examples of preferred antisense compounds useful in this invention include oligonucleotides containing modified backbones or non-natural internucleoside linkages. As defined in this specification, oligonucleotides having modified backbones include those that retain a phosphorus atom in the backbone and those that do not have a phosphorus atom in the backbone. For the purposes of this specification, and as sometimes referenced in the art, modified oligonucleotides that do not have a phosphorus atom in their internucleoside backbone can also be considered to be oligonucleosides.
[0023] Preferred modified oligonucleotide backbones include, for example, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkylphosphotriesters, methyl and other alkyl phosphonates including 3 'alkylene phosphonates and chiral phosphonates, phosphinates, phosphoramidates including 3 '-amino phosphoramidate and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotriesters, and boranophosphates having normal 3'-5' linkages, 2'-5' linked analogs of these, and those having inverted polarity wherein the adjacent pairs of nucleoside units are linked 3'-5' to 5'-3' or 2'-5' to 5'-2'. Various salts, mixed salts and free acid forms are also included.
[0024] Representative United States patents that teach the preparation of the above phosphorus-containing linkages include, but are not limited to, U.S.: 3,687,808; 4,469,863; 4,476,301 ; 5,023,243; 5,177,196; 5,188,897; 5,264,423; 5,276,019; 5,278,302; 5,286,717; 5,321,131 ; 5,399,676; 5,405,939; 5,453,496; 5,455,233; 5,466,677; 5,476,925; 5,519,126; 5,536,821; 5,541 ,306; 5,550,1 1 1; 5,563,253; 5,571,799; 5,587,361 ; and 5,625,050, each of which is herein incoφorated by reference. [0025] Preferred modified oligonucleotide backbones that do not include a phosphorus atom therein have backbones that are formed by short chain alkyl or cycloalkyl internucleoside linkages, mixed heteroatom and alkyl or cycloalkyl internucleoside linkages, or one or more short chain heteroatomic or heterocyclic internucleoside linkages. These include those having morpholino linkages (formed in part from the sugar portion of a nucleoside); siloxane backbones; sulfide, sulfoxide and sulfone backbones; formacetyl and thioformacetyl backbones; methylene formacetyl and thioformacetyl backbones; alkene containing backbones; sulfamate backbones; methyleneimino and methylenehydrazino backbones; sulfonate and sulfonamide backbones; amide backbones; and others having mixed N, O, S and CH component parts.
[0026] Representative United States patents that teach the preparation of the above oligonucleosides include, but are not limited to, U.S.: 5,034,506; 5,166,315; 5,185,444; 5,214,134; 5,216,141; 5,235,033; 5,264,562; 5,264,564; 5,405,938; 5,434,257; 5,466,677; 5,470,967; 5,489,677; 5,541 ,307; 5,561,225; 5,596,086; 5,602,240; 5,610,289; 5,602,240; 5,608,046; 5,610,289; 5,618,704; 5,623,070; 5,663,312; 5,633,360; 5,677,437; and 5,677,439, each of which is herein incorporated by reference.
[0027] In other preferred oligonucleotide mimetics, both the sugar and the internucleoside linkage, i.e., the backbone, of the nucleotide units are replaced with novel groups. The base units are maintained for hybridization with an appropriate nucleic acid target compound, one such oligomeric compound, an oligonucleotide mimetic that has been shown to have excellent hybridization properties, is referred to as a peptide nucleic acid (PNA). In PNA compounds, the sugar-backbone of an oligonucleotide is replaced with an amide containing backbone, in particular an aminoethylglycine backbone. The nucleobases are retained and are bound directly or indirectly to aza nitrogen atoms of the amide portion of the backbone. Representative United States patents that teach the preparation of PNA compounds include, but are not limited to, U.S.: 5,539,082; 5,714,331 ; and 5,719,262, each of which is herein incorporated by reference. Further teaching of PNA compounds can be found in Nielsen et al., Science, 1991 , 254, 1497-1500.
[0028] Most preferred embodiments of the invention are oligonucleotides with phosphorothioate backbones and oligonucleosides with heteroatom backbones, and in particular -CH2-NH-O-CH2-, -CH2-N (CH3) -O-CH2- [known as a methylene (methylimino) or MMI backbone] , - CH2-O-N (CH3) -CH2-, -CH2N(CH3)-N(CH3)- CH2- and -O-N(CH3)-CH -CH2- [wherein the native phosphodiester backbone is represented as -O-P-O-CH2-] of the above referenced U.S. patent 5,489,677, and the amide backbones of the above referenced U.S. patent 5,602,240. Also preferred are oligonucleotides having morpholino backbone structures of the above-referenced U.S. patent 5,034,506.
[0029] Modified oligonucleotides may also contain one or more substituted sugar moieties. Preferred oligonucleotides comprise one of the following at the 2' position: OH; F; O-, S-, or N-alkyl; O-, S-, or N-alkenyl; O-, S- or N-alkynyl; or O- alkyl-O-alkyl, wherein the alkyl, alkenyl and alkynyl may be substituted or unsubstituted C\ to Cι0 alkyl or C2 to Cio alkenyl and alkynyl. Particularly preferred are O[(CH2)nO]mCH3, O(CH2)n,OCH3, O(CH2)nNH2, O(CH2)nCH3, O(CH2)nONH2, and O(CH2nON[(CH2)nCH3)]2 where n and m are from 1 to about 10. Other preferred oligonucleotides comprise one of the following at the 2' position: Ci to Cio, ( lower alkyl, substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH, SCH3, OCN, Cl, Br, CN, CF3, OCF3, SOCH3, SO2CH3, ON02, NO2, N3, NH2, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino, substituted silyl, an RNA cleaving group, a reporter group, an intercalator, a group for improving the pharmacokinetic properties of an oligonucleotide, or a group for improving the pharmacodynamic properties of an oligonucleotide, and other substituents having similar properties. A preferred modification includes 2' - methoxyethoxy (2' -O-CH2CH2OCH3, also known as 2'-O- (2-methoxyethyl) or 2'- MOE) (Martin et al., Helv. Chim. Acta, 1995, 78, 486-504) i.e., an alkoxyalkoxy group. A further preferred modification includes 2'-dimethylaminooxyethoxy, i.e., a O(CH2)2ON(CH )2 group, also known as 2'-DMAOE, as described in examples herein below, and 2'-dimethylaminoethoxyethoxy (also known in the art as 2'-O- dimethyla inoethoxyethyl or 2'-DMAEOE), i.e., 2'-O-CH2-O-CH.-N (CH2)2, also described in examples herein below.
π [0030] Other preferred modifications include 2'-methoxy (2'-O CH3) , 2'- aminopropoxy (2'-O CH2 CH2 CH2NH2) and 2'-fluoro (2'-F). Similar modifications may also be made at other positions on the oligonucleotide, particularly the 3' position of the sugar on the 3' terminal nucleotide or in 2 '-5' linked oligonucleotides and the 5' position of 5' terminal nucleotide. Oligonucleotides may also have sugar mimetics such as cyclobutyl moieties in place of the pentofuranosyl sugar. Representative United States patents that teach the preparation of such modified sugar structures include, but are not limited to, U.S.: 4,981,957; 5,118,800; 5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786; 5,514,785; 5,519,134; 5,567,81 1 ; 5,576,427; 5,591 ,722; 5,597,909; 5,610,300; 5,627,053; 5,639,873; 5,646,265; 5,658,873; 5,670,633; and 5,700,920, each of which is herein incoφorated by reference in its entirety.
[0031] Oligonucleotides may also include nucleobase (often referred to in the art simply as "base") modifications or substitutions. As used herein, "unmodified" or "natural" nucleobases include the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C) and uracil (U). Modified nucleobases include other synthetic and natural nucleobases such as 5- methylcytosine (5-me-C), 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2- aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2- propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2- thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylquanine and 7-methyladenine, 8-azaguanine and 8- azaadenine, 7-deazaguanine and 7-deazaadenine and 3 -deaza guanine and 3- deazaadenine. Further nucleobases include those disclosed in United States Patent No. 3,687,808, those disclosed in The Concise Encyclopedia Of Polymer Science And Engineering, pages 858-859, Kroschwitz, J.I., ed. John Wiley & Sons, 1990, those disclosed by Englisch et al., Angewandte Chemie, International Edition, 1991 , 30, 613, and those disclosed by Sanghvi, Y.S., Chapter 15, Antisense Research and Applications, pages 289-302, Crooke, S.T. and Lebleu, B. ed., CRC Press, 1993. Certain of these nucleobases are particularly useful for increasing the binding affinity of the oligomeric compounds of the invention. These include 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and O-6 substituted purines, including 2-aminopropyladenine, 5-propynyluracil and 5-propynylcytosine. 5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2°C (Sanghvi, Y.S., Crooke, S.T. and Lebleu, B., eds, Antisense Research and Applications, CRC Press, Boca Raton, 1993, pp. 276-278) and are presently preferred base substitutions, even more particularly when combined with 2'-O- methoxyethyl sugar modifications. [0032] Representative United States patents that teach the preparation of certain of the above noted modified nucleobases as well as other modified nucleobases include, but are not limited to, the above noted U.S. 3,687,808, as well as U.S.: 4,845,205; 5,130,302; 5,134,066; 5,175,273; 5,367,066; 5,432,272; 5,457,187; 5,459,255; 5,484,908; 5,502,177; 5,525,71 1; 5,552,540; 5,587,469; 5,594,12', 5,596,091; 5,614,617; 5,750,692; and 5,681 ,941 , each of which is herein incoφorated by reference.
[0033] Another modification of the oligonucleotides of the invention involves chemically linking to the oligonucleotide one or more moieties or conjugates, which enhance the activity, cellular distribution or cellular uptake of the oligonucleotide. Such moieties include but are not limited to lipid moieties such as a cholesterol moiety (Letsinger et al., Proc. Natl. Acad. Sci. USA, 1989, 86, 6553-6556), cholic acid (Manoharan et al., Bioorg. Med. Chem. Let., 1994, 4, 1053-1060), a thioether, e.g., hexyl-S-tritylthiol (Manoharan et a\., Ann. N. Y. Acαd. Sci., 1992, 660, 306-309; Manoharan et al., Bioorg. Med. Chem. Let., 1993, 3, 2765-2770), a thiocholesterol (Oberhauser et al., Nucl. Acids Res., 1992, 20, 533-538), an aliphatic chain, e.g., dodecandiol or undecyl residues (Saison-Behmoaras et al., EMBO J., 1991 , 10, 1 11 1-1 1 18; 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 triethylammonium 1 ,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate (Manoharan et al., Tetrahedron Lett., 1995, 36, 365'-3654; Shea et al., Nucl. Acids Res., 1990, 18, 3777-3783), a polyamine or a polyethylene glycol chain (Mancharan et al., Nucleosides & Nucleotides, 1995, 14, 969-973), or adamantane acetic acid (Manoharan et al., Tetrahedron Lett., 1995, 36, 365'-3654), a palmityl moiety (Mishra et al., Biochim. Biophys. Acta, 1995, 1264, 229-237), or an octadecylamine or hexylamino-carbonyl-oxycholesterol moiety (Crooke et al., J Pharmacol. Exp. Ther., 1996, 277, 923-937).
[0034] Representative United States patents that teach the preparation of such oligonucleotide conjugates include, but are not limited to, U.S.: 4,828,979; 4,948,882; 5,218,105; 5,525,465; 5,541 ,313; 5,545,730; 5,552,538; 5,578,717, 5,580,731; 5,580,731 ; 5,591 ,584; 5,109,124; 5,1 18,802; 5,138,045; 5,414,077; 5,486,603; 5,512,439; 5,578,718; 5,608,046; 4,587,044; 4,605,735; 4,667,025; 4,762,779; 4,789,737; 4,824,941 ; 4,835,263; 4,876,335; 4,904,582; 4,958,013; 5,082,830; 5,112,963; 5,214,136; 5,082,830; 5,112,963; 5,214,136; 5,245,022; 5,254,469; 5,258,506; 5,262,536; 5,272,250; 5,292,873; 5,317,098; 5,371 ,241, 5,391,723; 5,416,203, 5,451,463; 5,510,475; 5,512,667; 5,514,785; 5,565,552; 5,567,810; 5,574,142; 5,585,481 ; 5,587,371 ; 5,595,726; 5,597,696; 5,599,923; 5,599,928 and 5,688,941 , each of which is herein incoφorated by reference. [0035] It is not necessary for all positions in a given compound to be uniformly modified, and in fact more than one of the aforementioned modifications may be incoφorated in a single compound or even at a single nucleoside within an oligonucleotide. The present invention also includes antisense compounds, which are chimeric compounds. "Chimeric" antisense compounds or "chimeras," in the context of this invention, are antisense compounds, particularly oligonucleotides, which contain two or more chemically distinct regions, each made up of at least one monomer unit, i.e., a nucleotide in the case of an oligonucleotide compound. These oligonucleotides typically contain at least one region wherein the oligonucleotide is modified so as to confer upon the oligonucleotide increased resistance to nuclease degradation, increased cellular uptake, and/or increased binding affinity for the target nucleic acid. An additional region of the oligonucleotide may serve as a substrate for enzymes capable of cleaving RNA:DNA or RNA:RNA hybrids. By way of example, RNase H is a cellular endonuclease, which cleaves the RNA strand of RNA:DNA duplex. Activation of RNase H, therefore, results in cleavage of the RNA target, thereby greatly enhancing the efficiency of oligonucleotide inhibition of gene expression. Consequently, comparable results can often be obtained with shorter oligonucleotides when chimeric oligonucleotides are used, compared to phosphorothioate deoxyoligonucleotides hybridizing to the same target region. Cleavage of the RNA target can be routinely detected by gel electrophoresis and, if necessary, associated nucleic acid hybridization techniques known in the art. [0036] Chimeric antisense compounds of the invention may be formed as composite structures of two or more oligonucleotides, modified oligonucleotides, oligonucleosides and/or oligonucleotide mimetics as described above. Such compounds have also been referred to in the art as hybrids or gapmers. Representative United States patents that teach the preparation of such hybrid structures include, but are not limited to, U.S.: 5,013,830; 5,149,797; 5,220,007; 5,256,775; 5,366,878; 5,403,71 1 ; 5,491,133; 5,565,350; 5,623,065; 5,652,355; 5,652,356; and 5,700,922, each of which is herein incoφorated by reference in its entirety.
[0037] The antisense compounds used in accordance with this invention may be conveniently, and routinely made through the well-known technique of solid phase synthesis. Equipment for such synthesis is sold by several vendors including, for example, Applied Biosystems (Foster City, CA). Any other means for such synthesis known in the art may additionally or alternatively be employed. It is well known to use similar techniques to prepare oligonucleotides such as the phosphorothioates and alkylated derivatives. [0038] The antisense compounds of the invention are synthesized in vitro and do not include antisense compositions of biological origin, or genetic vector constructs designed to direct the in vivo synthesis of antisense molecules. The compounds of the invention may also be admixed, encapsulated, conjugated or otherwise associated with other molecules, molecule structures or mixtures of compounds, as for example, liposomes, receptor targeted molecules, oral, rectal, topical or other formulations, for assisting in uptake, distribution and/or absoφtion. Representative United States patents that teach the preparation of such uptake, distribution and/or absoφtion assisting formulations include, but are not limited to, U.S.: 5,108,921 ; 5,354,844; 5,416,016; 5,459,127; 5,521 ,291 ; 5,543,158; 5,547,932; 5,583,020; 5,591 ,721 ; 4,426,330; 4,534,899; 5,013,556; 5,108,921 ; 5,213,804; 5,227,170; 5,264,221 ; 5,356,633; 5,395,619; 5,416,016; 5,417,978; 5,462,854; 5,469,854; 5,512,295; 5,527,528; 5,534,259; 5,543,152; 5,556,948; 5,580,575; and 5,595,756, each of which is herein incoφorated by reference. [0039] The antisense compounds of the invention encompass any pharmaceutically acceptable salts, esters, or salts of such esters, or any other compound which, upon administration to an animal including a human, is capable of providing (directly or indirectly) the biologically active metabolite or residue thereof. Accordingly, for example, the disclosure is also drawn to prodrugs and pharmaceutically acceptable salts of the compounds of the invention, pharmaceutically acceptable salts of such prodrugs, and other bioequivalents. [0040] The term "prodrug" indicates a therapeutic agent that is prepared in an inactive form that is converted to an active form (i.e., drug) within the body or cells thereof by the action of endogenous enzymes or other chemicals and/or conditions. In particular, prodrug versions of the oligonuclectides of the invention are prepared as SATE [(S-acetyl-2-thioethyl) phosphate] derivatives according to the methods disclosed in WO 93/24510 to Gosselin et al., published December 9, 1993 or in WO 94/26764 to Imbach et al. [0041] The term "pharmaceutically acceptable salts" refers to physiologically and pharmaceutically acceptable salts of the compounds of the invention: i.e., salts that retain the desired biological activity of the parent compound and do not impart undesired toxicological effects thereto. [0042] Pharmaceutically acceptable base addition salts are formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Examples of metals used as cations are sodium, potassium, magnesium, calcium, and the like. Examples of suitable amines are N, N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, dicyclohexylamine, ethylenediamine, N-methylglucamine, and procaine (see, for example, Berge et al., "Pharmaceutical Salts," J. of Pharma Sci., 1977, 66, 119). The base addition salts of said acidic compounds are prepared by contacting the free acid form with a sufficient amount of the desired base to produce the salt in the conventional manner. The free acid form may be regenerated by contacting the salt form with an acid and isolating the free acid in the conventional manner. The free acid forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free acid for puφoses of the present invention. As used herein, a "pharmaceutical addition salt" includes a pharmaceutically acceptable salt of an acid form of one of the components of the compositions of the invention. These include organic or inorganic acid salts of the amines. Preferred acid salts are the hydrochlorides, acetates, salicylates, nitrates and phosphates. Other suitable pharmaceutically acceptable salts are well known to those skilled in the art and include basic salts of a variety of inorganic and organic acids, such as, for example, with inorganic acids, such as for example hydrochloric acid, hydrobromic acid, sulfuric acid or phosphoric acid; with organic carboxylic, sulfonic, sulfo or phospho acids or N-substituted sulfamic acids, for example acetic acid, propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid, methylmaleic acid, fumaric acid, malic acid, tartaric acid, lactic acid, oxalic acid, gluconic acid, glucaric acid, glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-aminosalicylic acid, 2-phenoxybenzoic acid, 2- acetoxybenzoic acid, embonic acid, nicotinic acid or isonicotinic acid; and with amino acids, such as the 20 alpha-amino acids involved in the synthesis of proteins in nature, for example glutamic acid or aspartic acid, and also with phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, ethane-l,2-disulfonic acid, benzenesulfonic acid, 4-methylbenzenesulfoic acid, naphthalene-2-sulfonic acid, naphthalene- 1 ,5-disulfonic acid, 2- or 3- phosphogly cerate, glucose-6-phosphate, N-cyclohexylsulfamic acid (with the formation of cyclamates), or with other acid organic compounds, such as ascorbic acid. Pharmaceutically acceptable salts of compounds may also be prepared with a pharmaceutically acceptable cation. Suitable pharmaceutically acceptable cations are well known to those skilled in the art and include alkaline, alkaline earth, ammonium and quaternary ammonium cations. Carbonates or hydrogen carbonates are also possible. [0043] For oligonucleotides, preferred examples of pharmaceutically acceptable salts include but are not limited to (a) salts formed with cations such as sodium, potassium, ammonium, magnesium, calcium, polyamines such as spermine and spermidine, etc.; (b) acid addition salts formed with inorganic acids, for example hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid and the like; (c) salts formed with organic acids such as, for example, acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, methanesulfonic acid, p- toluenesulfonic acid, naphthalenedisulfonic acid, polygalacturonic acid, and the like; and (d) salts formed from elemental anions such as chlorine, bromine, and iodine.
[0044] The antisense compounds of the present invention can be utilized for diagnostics, therapeutics, prophylaxis and as research reagents and kits. For therapeutics, an animal, preferably a human, suspected of having a disease or disorder, which can be treated by modulating the expression of GR, is treated by administering antisense compounds in accordance with this invention. The compounds of the invention can be utilized in pharmaceutical compositions by adding an effective amount of an antisense compound to a suitable pharmaceutically acceptable diluent or carrier. Use of the antisense compounds and methods of the invention may also be useful prophylactically, e.g., to prevent or delay infection, inflammation or tumor formation, for example. [0045] The antisense compounds of the invention are useful for research and diagnostics, because these compounds hybridize to nucleic acids encoding GR, enabling sandwich and other assays to easily be constructed to exploit this fact. Hybridization of the antisense oligonucleotides of the invention with a nucleic acid encoding GR can be detected by means known in the art. Such means may include conjugation of an enzyme to the oligonucleotide, radiolabelling of the oligonucleotide or any other suitable detection means. Kits using such detection means for detecting the level of GR in a sample may also be prepared. [0046] The present invention also includes pharmaceutical compositions and formulations, which include the antisense compounds of the invention. The pharmaceutical compositions of the present invention may be administered in a number of ways depending upon whether local or systemic treatment is desired and upon the area to be treated. Administration may be topical (including ophthalmic and to mucous membranes including vaginal and rectal delivery), pulmonary, e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, intranasal, epidermal and transdermal), oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial, e.g., intrathecal or intraventricular, administration. Oligonucleotides with at least one 2'-O- methoxyethyl modification are believed to be particularly useful for oral administration.
[0047] Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable. Coated condoms, gloves and the like may also be useful. [0048] Compositions and formulations for oral administration include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets or tablets. Thickeners, flavoring agents, diluents, emulsifiers, dispersing aids or binders may be desirable.
[0049] Compositions and formulations for parenteral, intrathecal or intraventricular administration may include sterile aqueous solutions, which may also contain buffers, diluents and other suitable additives such as, but not limited to, penetration enhancers, carrier compounds and other pharmaceutically acceptable carriers or excipients.
[0050] Pharmaceutical compositions of the present invention include, but are not limited to, solutions, emulsions, and liposome-containing formulations. These compositions may be generated from a variety of components that include, but are not limited to, preformed liquids, self-emulsifying solids and self-emulsifying semisolids.
[0051] The pharmaceutical formulations of the present invention, which may conveniently be presented in unit dosage form, may be prepared according to conventional techniques well known in the pharmaceutical industry. Such techniques include the step of bringing into association the active ingredients with the pharmaceutical carrier(s) or excipient(s). In general the formulations are prepared by uniformly and intimately bringing into association the active ingredients with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product. [0052] The compositions of the present invention may be formulated into any of many possible dosage forms such as, but not limited to, tablets, capsules, liquid syrups, soft gels, suppositories, and enemas. The compositions of the present invention may also be formulated as suspensions in aqueous, non-aqueous or mixed media. Aqueous suspensions may further contain substances, which increase the viscosity of the suspension including, for example, sodium carboxymethylcellulose, sorbitol and/or dextran. The suspension may also contain stabilizers. [0053] In one embodiment of the present invention the pharmaceutical compositions may be formulated and used as foams. Pharmaceutical foams include formulations such as, but not limited to, emulsions, microemulsions, creams, jellies and liposomes. While basically similar in nature these formulations vary in the components and the consistency of the final product. The preparation of such compositions and formulations is generally known to those skilled in the pharmaceutical and formulation arts and may be applied to the formulation of the compositions of the present invention. Emulsions
[0054] The compositions of the present invention may be prepared and formulated as emulsions. Emulsions are typically heterogenous systems of one liquid dispersed in another in the form of droplets usually exceeding 0.1 μm in diameter. (Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199; Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., Volume 1, p. 245; Block in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 2, p. 335; Higuchi et al., in Remington 's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA, 1985, p. 301). Emulsions are often biphasic systems comprising of two immiscible liquid phases intimately mixed and dispersed with each other. In general, emulsions may be either water-in-oil (w/o) or of the oil-in-water (o/w) variety. When an aqueous phase is finely divided into and dispersed as minute droplets into a bulk oily phase the resulting composition is called a water-in-oil (w/o) emulsion. Alternatively, when an oily phase is finely divided into and dispersed as minute droplets into a bulk aqueous phase the resulting composition is called an oil-in-water (o/w) emulsion. Emulsions may contain additional components in addition to the dispersed phases and the active drug, which may be present as a solution in either the aqueous phase, oily phase or itself as a separate phase. Pharmaceutical excipients such as emulsifiers, stabilizers, dyes, and anti-oxidants may also be present in emulsions as needed. Pharmaceutical emulsions may also be multiple emulsions that are comprised of more than two phases such as, for example, in the case of oil-in-water-in-oil (o/w/o) and water-in- oil-in-water (w/o/w) emulsions. Such complex formulations often provide certain advantages that simple binary emulsions do not. Multiple emulsions in which individual oil droplets of an o/w emulsion enclose small water droplets constitute a w/o/w emulsion. Likewise a system of oil droplets enclosed in globules of water stabilized in an oily continuous provides an o/w/o emulsion. [0055] Emulsions are characterized by little or no thermodynamic stability. Often, the dispersed or discontinuous phase of the emulsion is well dispersed into the external or continuous phase and maintained in this form through the means of emulsifiers or the viscosity of the formulation. Either of the phases of the emulsion may be a semisolid or a solid, as is the case of emulsion-style ointment bases and creams. Other means of stabilizing emulsions entail the use of emulsifiers that may be incorporated into either phase of the emulsion. Emulsifiers may broadly be classified into four categories: synthetic surfactants, naturally occurring emulsifiers, absoφtion bases, and finely dispersed solids (Idson, in Pharmaceutical Dosaqe Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199).
[0056] Synthetic surfactants, also known as surface active agents, have found wide applicability in the formulation of emulsions and have been reviewed in the literature (Rieger, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 285; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), Marcel Dekker, Inc., New York, N.Y., 1988, volume 1, p. 199). Surfactants are typically amphiphilic and comprise a hydrophilic and a hydrophobic portion. The ratio of the hydrophilic to the hydrophobic nature of the surfactant has been termed the hydrophile/lipophile balance (HLB) and is a valuable tool in categorizing and selecting surfactants in the preparation of formulations. Surfactants may be classified into different classes based on the nature of the hydrophilic group: nonionic, anionic, cationic and amphoteric (Rieger, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1 , p. 285).
[0057] Naturally occurring emulsifiers used in emulsion formulations include lanolin, beeswax, phosphatides, lecithin and acacia. Absoφtion bases possess hydrophilic properties such that they can soak up water to form w/o emulsions yet retain their semisolid consistencies, such as anhydrous lanolin and hydrophilic petrolatum. Finely divided solids have also been used as good emulsifiers especially in combination with surfactants and in viscous preparations. These include polar inorganic solids, such as heavy metal hydroxides, nonswelling clays such as bentonite, attapulgite, hectorite, kaolin, montmorillonite, colloidal aluminum silicate and colloidal magnesium aluminum silicate, pigments and nonpolar solids such as carbon or glyceryl tristearate. [0058] A large variety of non-emulsifying materials are also included in emulsion formulations and contribute to the properties of emulsions. These include fats, oils, waxes, fatty acids, fatty alcohols, fatty esters, humectants, hydrophilic colloids, preservatives, and antioxidants (Block, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1 , p. 335; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1 , p. 199).
[0059] Hydrophilic colloids or hydrocolloids include naturally occurring gums and synthetic polymers such as polysaccharides (for example, acacia, agar, alginic acid, carrageenan, guar gum, karaya gum, and tragacanth), cellulose derivatives (for example, carboxymethyl cellulose and carboxypropylcellulose), and synthetic polymers (for example, carbomers, cellulose ethers, and carboxyvinyl polymers). These disperse or swell in water to form colloidal solutions that stabilize emulsions by forming strong interfacial films around the dispersedphase droplets and by increasing the viscosity of the external phase. [0060] Since emulsions often contain a number of ingredients such as carbohydrates, proteins, sterols and phosphatides that may readily support the growth of microbes, these formulations often incoφorate preservatives. Commonly used preservatives included in emulsion formulations include methyl paraben, propyl paraben, quaternary ammonium salts, benzalkonium chloride, esters of p- hydroxybenzoic acid, and boric acid. Antioxidants are also commonly added to emulsion formulations to prevent deterioration of the formulation. Antioxidants used may be free radical scavengers such as tocopherols, alkyl gallates, butylated hydroxyanisole, butylated hydroxytoluene, or reducing agents such as ascorbic acid and sodium metabisulfite, and antioxidant synergists such as citric acid, tartaric acid, and lecithin.
[0061] The application of emulsion formulations via dermatological, oral and parenteral routes and methods for their manufacture have been reviewed in the literature (Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1 , p. 199). Emulsion formulations for oral delivery have been very widely used because of reasons of ease of formulation, efficacy from an absoφtion and bioavailability standpoint. (Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1 , p. 245; Idson, in
Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199). Mineral-oil base laxatives, oil-soluble vitamins and high fat nutritive preparations are among the materials that have commonly been administered orally as o/w emulsions. [0062] In one embodiment of the present invention, the compositions of oligonucleotides and nucleic acids are formulated as microemulsions. A microemulsion may be defined as a system of water, oil and amphiphile, which is a single optically isotropic, and thermodynamically stable liquid solution (Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1 , p. 245). Typically microemulsions are systems that are prepared by first dispersing an oil in an aqueous surfactant solution and then adding a sufficient amount of a fourth component, generally an intermediate chain-length alcohol to form a transparent system. Therefore, microemulsions have also been described as thermodynamically stable, isotropically clear dispersions of two immiscible liquids that are stabilized by interfacial films of surface-active molecules (Leung and Shah, in: Controlled Release of Drugs: Polymers and Aggregate Systems, Rosoff, M., Ed., 1989, VCH Publishers, New York, pages 1852 '5). Microemulsions commonly are prepared via a combination of three to five components that include oil, water, surfactant, cosurfactant and electrolyte. Whether the microemulsion is of the water-in-oil (w/o) or an oil-in-water (o/w) type is dependent on the properties of the oil and surfactant used and on the structure and geometric packing of the polar heads and hydrocarbon tails of the surfactant molecules (Schott, in Remington 's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA, 1985, p. 271).
[0063] The phenomenological approach utilizing phase diagrams has been extensively studied and has yielded a comprehensive knowledge, to one skilled in the art, of how to formulate microemulsions (Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 245; Block, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1 , p. 335). Compared to conventional emulsions, microemulsions offer the advantage of solubilizing water-insoluble drugs in a formulation of thermodynamically stable droplets that are formed spontaneously. [0064] Surfactants used in the preparation of microemulsions include, but are not limited to, ionic surfactants, non-ionic surfactants, Brij 96, polyoxyethylene oleyl ethers, polyglycerol fatty acid esters, tetraglycerol monolaurate (ML310), tetraglycerol monooleate (MO310), hexaglycerol monooleate (PO310), hexaglycerol pentaoleate (PO500), decaglycerol monocaprate (MCA750), decaglycerol monooleate (MO750), decaglycerol sequioleate (S0750), decaglycerol decaoleate (DAO750), alone or in combination with cosurfactants. The cosurfactant, usually a short-chain alcohol such as ethanol, 1-propanol, and 1- butanol, serves to increase the interfacial fluidity by penetrating into the surfactant film and consequently creating a disordered film because of the void space generated among surfactant molecules. Microemulsions may, however, be prepared without the use of cosurfactants and alcohol-free self-emulsifying microemulsion systems are known in the art. The aqueous phase may typically be, but is not limited to, water, an aqueous solution of the drug, glycerol, PEG300, PEG400, polyglycerols, propylene glycols, and derivatives of ethylene glycol. The oil phase may include, but is not limited to, materials such as Captex 300, Captex 355, Capmul MCM, fatty acid esters, medium chain (C8-C12) mono, di, and triglycerides, polyoxyethylated glyceryl fatty acid esters, fatty alcohols, polyglycolized glycerides, saturated polyglycolized C8-C10 glycerides, vegetable oils and silicone oil.
[0065] Microemulsions are particularly of interest from the standpoint of drug solubilization and the enhanced absoφtion of drugs. Lipid based microemulsions (both o/w and w/o) have been proposed to enhance the oral bioavailability of drugs, including peptides (Constantinides et al., Pharmaceutical Research, 1994, 1 1, 1385-1390; Ritschel, Meth. Find. Exp. Clin. Pharmacol, 1993, 13, 205). Microemulsions afford advantages of improved drug solubilization, protection of drug from enzymatic hydrolysis, possible enhancement of drug absoφtion due to surfactant-induced alterations in membrane fluidity and permeability, ease of preparation, ease of oral administration over solid dosage forms, improved clinical potency, and decreased toxicity (Constantinides et al., Pharmaceutical Research, 1994, 11, 1385; Ho et al., J Pharm. Sci., 1996, 85, 138-143). Often microemulsions may form spontaneously when their components are brought together at ambient temperature. This may be particularly advantageous when formulating thermolabile drugs, peptides or oligonucleotides. Microemulsions have also been effective in the transdermal delivery of active components in both cosmetic and pharmaceutical applications. It is expected that the microemulsion compositions and formulations of the present invention will facilitate the increased systemic absoφtion of oligonucleotides and nucleic acids from the gastrointestinal tract, as well as improve the local cellular uptake of oligonucleotides and nucleic acids within the gastrointestinal tract, vagina, buccal cavity and other areas of administration. [0066] Microemulsions of the present invention may also contain additional components and additives such as sorbitan monostearate (Grill 3), Labrasol, and penetration enhancers to improve the properties of the formulation and to enhance the absoφtion of the oligonucleotides and nucleic acids of the present invention. Penetration enhancers used in the microemulsions of the present invention may be classified as belonging to one of five broad categories - surfactants, fatty acids, bile salts, chelating agents, and non-chelating non-surfactants (Lee et al., Critical
Reviews in Therapeutic Drug Carrier Systems, 1991 , p. 92). Each of these classes has been discussed above.
Liposomes [0067] There are many organized surfactant structures besides microemulsions that have been studied and used for the formulation of drugs. These include monolayers, micelles, bilayers and vesicles. Vesicles, such as liposomes, have attracted great interest because of their specificity and the duration of action they offer from the standpoint of drug delivery. As used in the present invention, the term "liposome" means a vesicle composed of amphiphilic lipids arranged in a spherical bilayer or bilayers.
[0068] Liposomes are unilamellar or multilamellar vesicles which have a membrane formed from a lipophilic material and an aqueous interior. The aqueous portion contains the composition to be delivered. Cationic liposomes possess the advantage of being able to fuse to the cell wall. Noncationic liposomes, although not able to fuse as efficiently with the cell wall, are taken up by macrophages in vivo. [0069] In order to cross intact mammalian skin, lipid vesicles must pass through a series of fine pores, each with a diameter less than 50 nm, under the influence of a suitable transdermal gradient. Therefore, it is desirable to use a liposome, which is highly deformable and able to pass through such fine pores. [0070] Further advantages of liposomes include; liposomes obtained from natural phospholipids are biocompatible and biodegradable; liposomes can incoφorate a wide range of water and lipid soluble drugs; liposomes can protect encapsulated drugs in their internal compartments from metabolism and degradation (Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, P. 245). Important considerations in the preparation of liposome formulations are the lipid surface charge, vesicle size and the aqueous volume of the liposomes. [0071] Liposomes are useful for the transfer and delivery of active ingredients to the site of action. Because the liposomal membrane is structurally similar to biological membranes, when liposomes are applied to a tissue, the liposomes start to merge with the cellular membranes. As the merging of the liposome and cell progresses, the liposomal contents are emptied into the cell where the active agent may act.
[0072] Liposomal formulations have been the focus of extensive investigation as the mode of delivery for many drugs. There is growing evidence that for topical administration, liposomes present several advantages over other formulations. Such advantages include reduced side-effects related to high systemic absoφtion of the administered drug, increased accumulation of the administered drug at the desired target, and the ability to administer a wide variety of drugs, both hydrophilic and hydrophobic, into the skin.
[0073] Several reports have detailed the ability of liposomes to deliver agents including high-molecular weight DNA into the skin. Compounds including analgesics, antibodies, hormones and high-molecular weight DNAs have been administered to the skin. The majority of applications resulted in the targeting of the upper epidermis.
[0074] Liposomes fall into two broad classes. Cationic liposomes are positively charged liposomes, which interact with the negatively charged DNA molecules to form a stable complex. The positively charged DNA/liposome complex binds to the negatively charged cell surface and is internalized in an endosome. Due to the acidic pH within the endosome, the liposomes are ruptured, releasing their contents into the cell cytoplasm (Wang et al., Biochem. Biophys. Res. Commun., 1987, 147, 980 - 985) [0075] Liposomes, which are pH-sensitive or negatively-charged, entrap DNA rather than complex with it. Since both the DNA and the lipid are similarly charged, repulsion rather than complex formation occurs. Nevertheless, some DNA is entrapped within the aqueous interior of these liposomes. pH-sensitive liposomes have been used to deliver DNA encoding the thymidine kinase gene to cell monolayers in culture. Expression of the exogenous gene was detected in the target cells (Zhou et al., Journal of Controlled Release, 1992, 19, 269-274). [0076] One major type of liposomal composition includes phospholipids other than naturally-derived phosphatidylcholine. Neutral liposome compositions, for example, can be formed from dimyristoyl phosphatidylcholine (DMPC) or dipalmitoyl phosphatidylcholine (DPPC). Anionic liposome compositions generally are formed from dimyristoyl phosphatidylglycerol, while anionic fusogenic liposomes are formed primarily from dioleoyl phosphatidylethanolamine (DOPE). Another type of liposomal composition is formed from phosphatidylcholine (PC) such as, for example, soybean PC, and egg PC. Another type is formed from mixtures of phospholipid and/or phosphatidylcholine and/or cholesterol.
[0077] Several studies have assessed the topical delivery of liposomal drug formulations to the skin. Application of liposomes containing interferon to guinea pig skin resulted in a reduction of skin heφes sores while delivery of interferon via other means (e.g. as a solution or as an emulsion) were ineffective (Weiner et al., Journal of Drug Targeting, 1992, 2, 405-410). Further, an additional study tested the efficacy of interferon administered as part of a liposomal formulation to the administration of interferon using an aqueous system, and concluded that the liposomal formulation was superior to aqueous administration (du Plessis et al., Antiviral Research, 1992, 18, 259-265).
[0078] Non-ionic liposomal systems have also been examined to determine their utility in the delivery of drugs to the skin, in particular systems comprising non-ionic surfactant and cholesterol. Non-ionic liposomal formulations comprising Novasome ™ I (glyceryl dilaurate/cholesterol/polyoxyethylene-10-stearyl ether) and Novasome™ II (glyceryl distearate/ cholesterol/polyoxyethylene-10-stearyl ether) were used to deliver cyclosporin-A into the dermis of mouse skin. Results indicated that such non-ionic liposomal systems were effective in facilitating the deposition of cyclosporin-A into different layers of the skin (Hu et al. S.TP.Pharma. Sci., 1994, 4, 6, 466).
[0079] Liposomes also include "sterically stabilized" liposomes, a term which, as used herein, refers to liposomes comprising one or more specialized lipids that, when incoφorated into liposomes, result in enhanced circulation lifetimes relative to liposomes lacking such specialized lipids. Examples of sterically stabilized liposomes are those in which part of the vesicle-forming lipid portion of the liposome (A) comprises one or more glycolipids, such as monosialoganglioside GM I , or (B) is derivatized with one or more hydrophilic polymers, such as a polyethylene glycol (PEG) moiety. While not wishing to be bound by any particular theory, it is thought in the art that, at least for sterically stabilized liposomes containing gangliosides, sphingomyelin, or PEG-derivatized lipids, the enhanced circulation half-life of these sterically stabilized liposomes derives from a reduced uptake into cells of the reticuloendothelial system (RES) (Allen et al., FEBS Letters, 1987, 223, 42; Wu et al., Cancer Research, 1993, 53, 3765). [0080] Various liposomes comprising one or more glycolipids are known in the art. Papahadjopoulos et al. (Ann. N.Y. Acad. Sci, 1987, 507, 64) reported the ability of monosialoganglioside GM1, galactocerebroside sulfate and phosphatidylinositol to improve blood half-lives of liposomes. These findings were expounded upon by Gabizon et al. (Proc. Natl. Acad. Sci. U.S.A., 1988, 85, 6949). U.S. Patent No. 4,837,028 and WO 88/04924, both to Allen et al., disclose liposomes comprising (1) sphingomyelin and (2) the ganglioside Gjor a galactocerebroside sulfate ester. U.S. Patent No. 5,543,152 (Webb et al.) discloses liposomes comprising sphingomyelin. Liposomes comprising 1 ,2-sn-dimyristoylphosphatidylcholine are disclosed in WO 97/13499 (Lim et al.).
[0081] Many liposomes comprising lipids derivatized with one or more hydrophilic polymers, and methods of preparation thereof, are known in the art. Sunamoto et al. (Bull. Chem. Soc. Jpn., 1980, 53, 2778) described liposomes comprising a nonionic detergent, 2C1215G, that contains a PEG moiety. Ilium et al. (FEBS Lett., 1984, 167, 79) noted that hydrophilic coating of polystyrene particles with polymeric glycols results in significantly enhanced blood half-lives. Synthetic phospholipids modified by the attachment of carboxylic groups of polyalkylene glycols (e.g., PEG) are described by Sears (U.S. Patent Nos. 4,426,330 and 4,534,899). Klibanov et al. (FEBS Lett., 1990, 268, 235) described experiments demonstrating that liposomes comprising phosphatidylethanolamine (PE) derivatized with PEG or PEG stearate have significant increases in blood circulation half-lives. Blume et al. (Biochimica et Biophysica Acta, 1990, 1029, 91) extended such observations to other PEGderivatized phospholipids, e.g., DSPE-PEG, formed from the combination of distearoylphosphatidylethanolamine (DSPE) and PEG. Liposomes having covalently bound PEG moieties on their external surface are described in European Patent No. EP 0445 131 Bl and WO 90/04384 to Fisher. Liposome compositions containing 1-20 mole percent of PE derivatized with PEG, and methods of use thereof, are described by Woodle et al. (U.S. Patent Nos. 5,013,556 and 5,356,633) and Martin et al. (U.S. Patent No. 5,213,804 and
European Patent No. EP 0 496 813 Bl). Liposomes comprising a number of other lipid-polymer conjugates are disclosed in WO 91/05545 and U.S. Patent No. 5,225,212 (both to Martin et al.) and in WO 94/20073 (Zalipsky et al.) Liposomes comprising PEG-modified ceramide lipids are described in WO 96/10391 (Choi et al.). U.S. Patent Nos. 5,540,935 (Miyazaki et al.) and 5,556,948 (Tagawa et al.) describe PEG-containing liposomes that can be further derivatized with functional moieties on their surfaces. [0082] A limited number of liposomes comprising nucleic acids are known in the art. WO 96/40062 to Thierry et al. discloses methods for encapsulating high molecular weight nucleic acids in liposomes. U.S. Patent No. 5,264,221 to Tagawa et al. discloses protein-bonded liposomes and asserts that the contents of such liposomes may include an antisense RNA. U.S. Patent No. 5,665,710 to Rahman et al. describes certain methods of encapsulating oligodeoxynucleotides in liposomes. WO 97/04787 to Love et al. discloses liposomes comprising antisense oligonucleotides targeted to the raf gene. [0083] Transfersomes are yet another type of liposomes, and are highly deformable lipid aggregates which are attractive candidates for drug delivery vehicles. Transfersomes may be described as lipid droplets which are so highly deformable that they are easily able to penetrate through pores which are smaller than the droplet. Transfersomes are adaptable to the environment in which they are used, e.g. they are self-optimizing (adaptive to the shape of pores in the skin), self- repairing, frequently reach their targets without fragmenting, and often self-loading. To make transfersomes it is possible to add surface edge-activators, usually surfactants, to a standard liposomal composition. Transfersomes have been used to deliver serum albumin to the skin. The transfersome-mediated delivery of serum albumin has been shown to be as effective as subcutaneous injection of a solution containing serum albumin.
[0084] Surfactants find wide application in formulations such as emulsions (including microemulsions) and liposomes. The most common way of classifying and ranking the properties of the many different types of surfactants, both natural and synthetic, is by the use of the hydrophile/lipophile balance (HLB). The nature of the hydrophilic group (also known as the "head") provides the most useful means for categorizing the different surfactants used in formulations (Rieger, in Pharmaceutical Dosage Forms, Marcel Dekker, Inc., New York, NY, 1988, p. 285) [0085] If the surfactant molecule is not ionized, it is classified as a nonionic surfactant. Nonionic surfactants find wide application in pharmaceutical and cosmetic products and are usable over a wide range of pH values. In general their HLB values range from 2 to about 18 depending on their structure. Nonionic surfactants include nonionic esters such as ethylene glycol esters, propylene glycol esters, glyceryl esters, polyglyceryl esters, sorbitan esters, sucrose esters, and ethoxylated esters. Nonionic alkanolamides and ethers such as fatty alcohol ethoxylates, propoxylated alcohols, and ethoxylated/propoxylated block polymers are also included in this class. The polyoxyethylene surfactants are the most popular members of the nonionic surfactant class. |0086] If the surfactant molecule carries a negative charge when it is dissolved or dispersed in water, the surfactant is classified as anionic. Anionic surfactants include carboxylates such as soaps, acyl lactylates, acyl amides of amino acids, esters of sulfuric acid such as alkyl sulfates and ethoxylated alkyl sulfates, sulfonates such as alkyl benzene sulfonates, acyl isethionates, acyl taurates and sulfosuccinates, and phosphates. The most important members of the anionic surfactant class are the alkyl sulfates and the soaps.
[0087] If the surfactant molecule carries a positive charge when it is dissolved or dispersed in water, the surfactant is classified as cationic. Cationic surfactants include quaternary ammonium salts and ethoxylated amines. The quaternary ammonium salts are the most used members of this class.
[0088] If the surfactant molecule has the ability to carry either a positive or negative charge, the surfactant is classified as amphoteric. Amphoteric surfactants include acrylic acid derivatives, substituted alkylamides, N-alkylbetaines and phosphatides. [0089] The use of surfactants in drug products, formulations and in emulsions has been reviewed (Rieger, in Pharmaceutical Dosage Forms, Marcel Dekker, Inc., New York, NY, 1988, p. 285). Penetration Enhancers
[0090] In one embodiment, the present invention employs various penetration enhancers to effect the efficient delivery of nucleic acids particularly oligonucleotides, to the skin of animals. Most drugs are present in solution in both ionized and nonionized forms. However, usually only lipid soluble or lipophilic drugs readily cross cell membranes. It has been discovered that even non-1 ipophilic drugs may cross cell membranes if the membrane to be crossed is treated with a penetration enhancer. In addition to aiding the diffusion of non-lipophilic drugs across cell membranes, penetration enhancers also enhance the permeability of lipophilic drugs.
10091] Penetration enhancers may be classified as belonging to one of five broad categories, i.e., surfactants, fatty acids, bile salts, chelating agents, and non- chelating nonsurfactants (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991 , p.92). Each of the above mentioned classes of penetration enhancers are described below in greater detail.
Surfactants
[0092] In connection with the present invention, surfactants (or "surface-active agents") are chemical entities which, when dissolved in an aqueous solution, reduce the surface tension of the solution or the interfacial tension between the aqueous solution and another liquid, with the result that absoφtion of oligonucleotides through the mucosa. is enhanced. In addition to bile salts and fatty acids, these penetration enhancers include, for example, sodium lauryl sulfate, polyoxyethylene- 9-lauryl ether and polyoxyethylene-20-cetyl ether) (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, p.92); and perfluorochemical emulsions, such as FC-43. Takahashi et al., J. Pharm. Pharmacol, 1988, 40, 252).
Fatty acids
[0093] Various fatty acids and their derivatives which act as penetration enhancers include, for example, oleic acid, lauric acid, capric acid (n-decanoic acid), myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, dicaprate, tricaprate, monoolein (l-monooleoyl-.rac-glycerol), dilaurin, caprylic acid, arachidonic acid, glycerol 1 -monocaprate, 1 -dodecylazacycloheptan-2-one, acylcarnitines, acylcholines, C,.ιo alkyl esters thereof (e.g., methyl, isopropyl and t- butyl), and mono- and di-glycerides thereof (i.e., oleate, laurate, caprate, myristate, palmitate, stearate, linoleate, etc.) (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, p.92; Muranishi, Critical Reviews in Therapeutic Drug Carrier Systems, 1990, 7, 1-33; El Hariri et al., J. Pharm. Pharmacol, 1992, 44, 651 -654).
Bile salts [0094] The physiological role of bile includes the facilitation of dispersion and absoφtion of lipids and fat-soluble vitamins (Brunton, Chapter 38 in: Goodman & Gilman's The Pharmacological Basis of Therapeutics, 9th Ed., Hardman et al. Eds. McGraw-Hill, New York, 1996, pp. 934-935). Various natural bile salts, and their synthetic derivatives, act as penetration enhancers. Thus the term "bile salts" includes any of the naturally occurring components of bile as well as any of their synthetic derivatives. The bile salts of the invention include, for example, cholic acid (or its pharmaceutically acceptable sodium salt, sodium cholate), dehydrocholic acid (sodium dehydrocholate), deoxycholic acid (sodium deoxycholate), glucholic acid (sodium glucholate), glycholic acid (sodium glycocholate), glycodeoxycholic acid (sodium glycodeoxycholate), taurocholic acid (sodium taurocholate), taurodeoxycholic acid (sodium taurodeoxycholate), chenodeoxycholic acid (sodium chenodeoxycholate), ursodeoxycholic acid (UDCA), sodium tauro-24,25-dihydro-fusidate (STDHF), sodium glycodihydrofusidate'and polyoxyethylene-9-lauryl ether (POE) (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, page 92; Swinyard, Chapter 39 In: Remington 's Pharmaceutical Sciences, 18th Ed., Gennaro, ed., Mack Publishing Co., Easton, PA, 1990, pages 782-783; Muranishi, Critical Reviews in Therapeutic Drug Carrier Systems, 1990, 7, 1 -33; Yamamoto et al., J. Pharm. Exp. Ther., 1992, 263, 25; Yamashita et al., J. Pharm. Sci., 1990, 79, 579-583).
Chelating Agents
[0095] Chelating agents, as used in connection with the present invention, can be defined as compounds that remove metallic ions from solution by forming complexes therewith, with the result that absoφtion of oligonucleotides through the mucosa is enhanced. With regards to their use as penetration enhancers in the present invention, chelating agents have the added advantage of also serving as DNase inhibitors, as most characterized DNA nucleases require a divalent metal ion for catalysis and are thus inhibited by chelating agents (Jarrett, J. Chromatogr., 1993, 618, 315-339). Chelating agents of the invention include but are not limited to disodium. ethylenediaminetetraacetate (EDTA), citric acid, salicylates (e.g., sodium salicylate, 5-methoxysalicylate and homovanilate), N-acyl derivatives of collagen, laureth-9 and N-amino acyl derivatives of beta-diketones (enamines)(Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, page 92;
Muranishi, Critical Reviews in Therapeutic Drug Carrier Systems, 1990, 7, 1-33; Buur et al., J Control Rel, 1990, 14, 43-51). Non-chelating non-surfactants
[0096] As used herein, nonchelating non-surfactant penetration enhancing compounds can be defined as compounds that demonstrate insignificant activity as chelating agents or as surfactants but that nonetheless enhance absoφtion of oligonucleotides through the alimentary mucosa (Muranishi, Critical Reviews in Therapeutic Drug Carrier Systems, 1990, 7, 1-33). This class of penetration enhancers include, for example, unsaturated cyclic ureas, 1 -alkyl- and 1- alkenylazacyclo-alkanone derivatives (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991 , page 92); and non-steroidal anti-inflammatory agents such as diclofenac sodium, indomethacin and phenylbutazone (Yamashita et al., J. Pharm. Pharmacol, 1987, 39, 621-626).
[0097] Agents that enhance uptake of oligonucleotides at the cellular level may also be added to the pharmaceutical and other compositions of the present invention. For example, cationic lipids, such as lipofectin (Junichi et al, U.S. Patent No. 5,705,188), cationic glycerol derivatives, and polycationic molecules, such as polylysine (Lollo et al., PCT Application WO 97/30731), are also known to enhance the cellular uptake of oligonucleotides.
[0098] Other agents may be utilized to enhance the penetration of the administered nucleic acids, including glycols such as ethylene glycol and propylene glycol, pyrrols such as 2-pyrrol, azones, and teφenes such as limonene and menthone.
Carriers
[0099] Certain compositions of the present invention also incoφorate carrier compounds in the formulation. As used herein, "carrier compound" or "carrier" can refer to a nucleic acid, or analog thereof, which is inert (i.e., does not possess biological activity per se) but is recognized as a nucleic acid by in vivo processes that reduce the bioavailability of a nucleic acid having biological activity by, for example, degrading the biologically active nucleic acid or promoting its removal from circulation. The coadministration of a nucleic acid and a carrier compound, typically with an excess of the latter substance, can result in a substantial reduction of the amount of nucleic acid recovered in the liver, kidney or other extracirculatory reservoirs, presumably due to competition between the carrier compound and the nucleic acid for a common receptor. For example, the recovery of a partially phosphorothioate oligonuclectide in hepatic tissue can be reduced when it is coadministered with polyinosinic acid, dextran sulfate, polycytidic acid or 4- acetamido-4'isothiocyano-stilbene-2,2'disulfonic acid (Miyao et al., Antisense Res. Dev., 1995, 5, 1 15-121 ; Takakura et al, Antisense & Nucl. Acid Drug Dev., 1996, 6, 177-183).
Excipients
[00100] In contrast to a carrier compound, a "pharmaceutical carrier" or "excipient" is a pharmaceutically acceptable solvent, suspending agent or any other pharmacologically inert vehicle for delivering one or more nucleic acids to an animal. The excipient may be liquid or solid and is selected, with the planned manner of administration in mind, so as to provide for the desired bulk, consistency, etc., when combined with a nucleic acid and the other components of a given pharmaceutical composition. Typical pharmaceutical carriers include, but are not limited to, binding agents (e.g., pregelatinized maize starch, polyvinylpyrrolidone or hydroxypropyl methylcellulose, etc.); fillers (e.g., lactose and other sugars, microcrystalline cellulose, pectin, gelatin, calcium sulfate, ethyl cellulose, polyacrylates or calcium hydrogen phosphate, etc.); lubricants (e.g., magnesium stearate, talc, silica, colloidal silicon dioxide, stearic acid, metallic stearates, hydrogenated vegetable oils, corn starch, polyethylene glycols, sodium benzoate, sodium acetate, etc.); disintegrants (e.g., starch, sodium starch glycolate, etc.); and wetting agents (e.g., sodium lauryl sulphate, etc.). |00101] Pharmaceutically acceptable organic or inorganic excipient suitable for non-parenteral administration which do not deleteriously react with nucleic acids can also be used to formulate the compositions of the present invention. Suitable pharmaceutically acceptable carriers include, but are not limited to, water, salt solutions, alcohols, polyethylene glycols, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethyl cellulose, polyvinylpyrrolidone and the like.
|00102] Formulations for topical administration of nucleic acids may include sterile and non-sterile aqueous solutions, non-aqueous solutions in common solvents such as alcohols, or solutions of the nucleic acids in liquid or solid oil bases. The solutions may also contain buffers, diluents and other suitable additives. Pharmaceutically acceptable organic or inorganic excipients suitable for non- parenteral administration which do not deleteriously react with nucleic acids can be used. [00103] Suitable pharmaceutically acceptable excipients include, but are not limited to, water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethyl cellulose, polyvinylpyrrolidone and the like. Other Components [00104] The compositions of the present invention may additionally contain other adjunct components conventionally found in pharmaceutical compositions, at their art-established usage levels. Thus, for example, the compositions may contain additional, compatible, pharmaceutically-active materials such as, for example, antipruritics, astringents, local anesthetics or anti -inflammatory agents, or may contain additional materials useful in physically formulating various dosage forms of the compositions of the present invention, such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickening agents and stabilizers. However, such materials, when added, should not unduly interfere with the biological activities of the components of the compositions of the present invention.' The formulations can be sterilized and, if desired, mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorings, flavorings and/or aromatic substances and the like which do not deleteriously interact with the nucleic acid(s) of the formulation. [00105] Aqueous suspensions may contain substances which increase the viscosity of the suspension including, for example, sodium carboxymethyl cellulose, sorbitol and/or dextran. The suspension may also contain stabilizers.
[00106] Certain embodiments of the invention provide pharmaceutical compositions containing (a) one or more antisense compounds and (b) one or more other chemotherapeutic agents which function by a non-antisense mechanism. Examples of such chemotherapeutic agents include, but are not limited to, anticancer drugs such as daunorubicin, dactinomycin, doxorubicin, bleomycin, mitomycin, nitrogen mustard, chlorambucil, melphalan, cyclophosphamide, 6- mercaptopurine, 6-thioguanine, cytarabine (CA), 5-fluorouracil (5-FU), floxuridine (5-FUdR), methotrexate (MTX), colchicine, vincristine, vinblastine, etoposide, teniposide, cisplatin and diethylstilbestrol (DES). See, generally, The Merck Manual of Diagnosis and Therapy, 15th Ed., Berkow et al., eds., 1987, Rahway, N.J., pages 1206-1228). Anti-inflammatory drugs, including but not limited to nonsteroidal anti-inflammatory drugs and corticosteroids, and antiviral drugs, including but not limited to ribivirin, vidarabine, acyclovir and ganciclovir, may also be combined in compositions of the invention. See, generally, The Merck Manual of Diagnosis and Therapy, 15th Ed., Berkow et al., eds., 1987, Rahway, N.J., pages 2499-2506 and 46-49, respectively), other non-antisense chemotherapeutic agents are also within the scope of this invention. Two or more combined compounds may be used together or sequentially. [00107] In another related embodiment, compositions of the invention may contain one or more antisense compounds, particularly oligonucleotides, targeted to a first nucleic acid and one or more additional antisense compounds targeted to a second nucleic acid target. Numerous examples of antisense compounds are known in the art. Two or more combined compounds may be used together or sequentially. [00108] The formulation of therapeutic compositions and their subsequent administration is believed to be within the skill of those in the art. Dosing is dependent on severity and responsiveness of the disease state to be treated, with the course of treatment lasting from several days to several months, or until a cure is effected or a diminution of the disease state is achieved. Optimal dosing schedules can be calculated from measurements of drug accumulation in the body of the patient. Persons of ordinary skill can easily determine optimum dosages, dosing methodologies and repetition rates. Optimum dosages may vary depending on the relative potency of individual oligonucleotides, and can generally be estimated based on EC5os found to be effective in in vitro and in vivo animal models. In general, dosage is from 0.01 μg to 100 g per kg of body weight, and may be given once or more daily, weekly, monthly or yearly, or even once every 2 to 20 years. Persons of ordinary skill in the art can easily estimate repetition rates for dosing based on measured residence times and concentrations of the drug in bodily fluids or tissues. Following successful treatment, it may be desirable to have the patient undergo maintenance therapy to prevent the recurrence of the disease state, wherein the oligonucleotide is administered in maintenance doses, ranging from 0.01 μg to 100 g per kg of body weight, once or more daily, to once every 20 years.
[00109] The complete content of all publications, patents, and patent applications cited in this disclosure are herein incoφorated by reference as if each individual publication, patent, or patent application were specifically and individually indicated to be incoφorated by reference.
[00110] Although the foregoing invention has been described in some detail by way of illustration and example for the purposes of clarity of understanding, it will be readily apparent to one skilled in the art in light of the teachings of this invention that changes and modifications can be made without departing from the spirit and scope of the present invention. The following examples are provided for exemplification puφoses only and are not intended to limit the scope of the invention, which has been described in broad terms above.
EXAMPLES
Example 1 Nucleoside Phosphoramidites for Oligonucleotide Synthesis Deoxy and 2'- alkoxy amiditcs
[00111] 2'-Deoxy and 2'-methoxy beta-cyanoethyldiisopropyl phosphoramidites are available from commercial sources (e.g. Chemgenes, Needham MA or Glen Research, Inc. Sterling VA). Other 2'-O-alkoxy substituted nucleoside amidites are prepared as described in U.S. Patent 5,506,351, herein incoφorated by reference. For oligonucleotides synthesized using 2 '-alkoxy amidites, the standard cycle for unmodified oligonucleotides is utilized, except the wait step after pulse delivery of tetrazole and base is increased to 360 seconds. |00112] Oligonucleotides containing 5-methyl-2'-deoxycytidine (5-Me-C) nucleotides are synthesized according to published methods [Sanghvi, et. al., Nucleic Acids Research, 1993, 21 , 3197-3203] using commercially available phosphoramidites (Glen Research, Sterling VA or ChemGenes, Needham MA). 2'-Fluoro amidites 2'-Fluorodeoxyadenosine amidites
[00113] 2'-fluoro oligonucleotides are synthesized as described previously [Kawasaki, et. al., J. Med. Chem., 1993, 36, 831-841 ] and United States patent 5,670,633, herein incoφorated by reference. Briefly, the protected nucleoside N6- benzoyl-2'-deoxy-2'-fluoroadenosine is synthesized utilizing commercially available 9-beta-D-arabinofuranosyladenine as starting material and by modifying literature procedures whereby the 2'-alpha-fluoro atom is introduced by a SN2- displacement of a 2'-beta-trityl group. Thus N6-benzoyl-9-beta-D- arabinofuranosyladenine is selectively protected in moderate yield as the 3 ',5'- ditetrahydropyranyl (THP) intermediate. Deprotection of the THP and N6-benzoyl groups is accomplished using standard methodologies and standard methods are used to obtain the 5'-dimethoxytrityl-(DMT) and 5'-DMT-3'-phosphoramidite intermediates. 2'-Fluorodeoxyguanosine
[00114] The synthesis of 2'-deoxy-2'-fluoroguanosine is accomplished using tetraisopropyldisiloxanyl (TPDS) protected 9-beta-D-arabinofuranosylguanine as starting material, and conversion to the intermediate diisobutyrylarabinofuranosylguanosine. Deprotection of the TPDS group is followed by protection of the hydroxyl group with THP to give diisobutyryl di-THP protected arabinofuranosylguanine. Selective O-deacylation and triflation is followed by treatment of the crude product with fluoride, then deprotection of the THP groups. Standard methodologies are used to obtain the 5'-DMT- and 5'-DMT- 3 '-phosphoramidites. 2'-Fluorouridine
[00115] Synthesis of 2 '-deoxy-2 '-fluorouridine is accomplished by the modification of a literature procedure in which 2,2'anhydro-l-beta-D- arabinofuranosyluracil is treated with 70% hydrogen fluoride-pyridine. Standard procedures are used to obtain the 5'-DMT and 5'-DMT-3'-phosphoramidites. 2'-Fluorodeoxycytidine
[00116] 2 '-deoxy-2 '-fluorocytidine is synthesized via amination of 2'-deoxy-2'- fluorouridine, followed by selective protection to give N4-benzoyl-2 '-deoxy-2 '- fluorocytidine. Standard procedures are used to obtain the 5'-DMT and 5'-DMT- 3 'phosphoramidites.
2'-O-(2-Methoxyethyl) modified amidites
[00117] 2'-O-Methoxyethyl-substituted nucleoside amidites are prepared as follows, or alternatively, as per the methods of Martin, P., Helvetica Chimica Acta, 1995, 78, 486-504.
2,2'-Anhydro[]-(beta-D-arabinofuranosyl)-5-methyluridinel [00118] 5-Methyluridine (ribosylthymine, commercially available through Yamasa, Choshi, Japan) (72.0 g, 0.279 M), diphenyl carbonate (90.0 g, 0.420 M) and sodium bicarbonate (2.0 g, 0.024 M) are added to DMF (300 mL). The mixture is heated to reflux, with stirring, allowing the evolved carbon dioxide gas to be released in a controlled manner. After 1 hour, the slightly darkened solution is concentrated under reduced pressure. The resulting syrup is poured into diethylether (2.5 L), with stirring. The product formed a gum. The ether is decanted and the residue is dissolved in a minimum amount of methanol (ca. 400 mL). The solution is poured into fresh ether (2.5 L) to yield a stiff gum. The ether is decanted and the gum is dried in a vacuum oven (60°C at 1 mm Hg for 24 h) to give a solid that is crushed to a light tan powder. The material is used as is for further reactions (or it can be purified further by column chromatography using a gradient of methanol in ethyl acetate (10-25%) to give a white solid. 2'-O-MethoxyethyI-5-methyluridine
[00119] 2,2'-Anhydro-5-methyluridine (195 g, 0.81 M), tris(2- methoxyethyl)borate (231 g, 0.98 M) and 2-methoxyethanol (1.2 L) are added to a 2 L stainless steel pressure vessel and placed in a pre -heated oil bath at 160°C. After heating for 48 hours at 155-160°C, the vessel is opened and the solution evaporated to dryness and triturated with MeOH (200 mL). The residue is suspended in hot acetone (1 L). The insoluble salts are filtered, washed with acetone (150 mL) and the filtrate evaporated. The residue (280 g) is dissolved in CH3CN (600 mL) and evaporated. A silica gel column (3 kg) is packed in CH2C12 /acetone /MeOH (20:5:3) containing 0.5% Et3NH. The residue is dissolved in CH2C12 (250 mL) and adsorbed onto silica (150 g) prior to loading onto the column. The product is eluted with the packing solvent to give the title product. Additional material can be obtained by reworking impure fractions. 2'-O-Methoxyethy]-5'-O-dimethoxytπtyl-5-methy-uridine
|00120] 2'-O-Methoxyethyl-5-methyluridine (160 g, 0.506 M) is co-evaporated with pyridine (250 mL) and the dried residue dissolved in pyridine (1.3 L). A first aliquot of dimethoxytrityl chloride (94.3 g, 0.278 M) is added and the mixture stirred at room temperature for one hour. A second aliquot of dimethoxytrityl chloride (94.3 g, 0.278 M) is added and the reaction stirred for an additional one hour. Methanol (170 mL) is then added to stop the reaction.The solvent is evaporated and triturated with CH3CN (200 mL) The residue is dissolved in CHC1 (1.5 L) and extracted with 2x500 mL of saturated NaHCO3 and 2x500 mL of saturated NaCl. The organic phase is dried over Na2SO4, filtered, and evaporated. The residue is purified on a 3.5 kg silica gel column, packed and eluted with EtOAc/hexane/ acetone (5:5:1) containing 0-5% Et NH. The pure fractions are evaporated to give the title product. 3'-O-Acetyl-2'-O-methoxyethyl-5'-O-dimethoxytrityl-5-methy]uridine 100121] 2'-O-Methoxyethyl-5'-O-dimethoxytrityl-5-methyluridine (106 g, 0.167 M), DMF/pyridine (750 mL of a 3:1 mixture prepared from 562 mL of DMF and 188 mL of pyridine) and acetic anhydride (24.38 mL, 0.258 M) are combined and stirred at room temperature for 24 hours. The reaction is monitored by TLC by first quenching the TLC sample with the addition of MeOH. Upon completion of the reaction, as judged by TLC, MeOH (50 mL) is added and the mixture evaporated at 35°C. The residue is dissolved in CHC13 (800 mL) and extracted with 2x200 mL of saturated sodium bicarbonate and 2x200 mL of saturated NaCl. The water layers are back extracted with 200 mL of CHCI3. The combined organics are dried with sodium sulfate and evaporated to a residue. The residue is purified on a 3.5 kg silica gel column and eluted using EtOAc/hexane(4:l). Pure product fractions are evaporated to yield the title compounds.
3'-O-Acetyl-2'-O-methoxyethyl-5'-O-dimethoxytrityl-5-methyl-4- triazoleuridine [00122] A first solution is prepared by dissolving 3'-O-acetyl-2'-O- methoxyethyl-5 '-O-dimethoxytrityl-5-methyluridine (96 g, 0.144 M) in CH3CN (700 mL) and set aside. Triethylamine (189 mL, 1.44 M) is added to a solution of triazole (90 g, 1.3 M) in CH3CN (1 L), cooled to -5°C and stirred for 0.5 h using an overhead stirrer. POCl3 is added dropwise, over a 30 minute period, to the stirred solution maintained at 0-10°C, and the resulting mixture stirred for an additional 2 hours. The first solution is added dropwise, over a 45 minute period, to the latter solution. The resulting reaction mixture is stored overnight in a cold room. Salts are filtered from the reaction mixture and the solution is evaporated. The residue is dissolved in EtOAc (1 L) and the insoluble solids are removed by filtration. The filtrate is washed with 1x300 mL of NaHCO3 and 2x300 mL of saturated NaCl, dried over sodium sulfate and evaporated. The residue is triturated with EtOAc to give the title compound. 2'-O-Methoxyethyl-5'-O-dimethoxytrityl-5-methylcytidine [00123] A solution of 3'-O-acetyl-2'-O-methoxyethyl-5'-O-dimethoxytrityl-5- methyl-4-triazoleuridine (103 g, 0.141 M) in dioxane (500 mL) and NH4OH (30 mL) is stirred at room temperature for 2 hours. The dioxane solution is evaporated and the residue azeotroped with MeOH (2x200 mL). The residue is dissolved in MeOH (300 mL) and transferred to a 2 liter stainless steel pressure vessel. MeOH (400 mL) saturated with NH3 gas is added and the vessel heated to 100°C for 2 hours (TLC showed complete conversion). The vessel contents are evaporated to dryness and the residue is dissolved in EtOAc (500 mL) and washed once with saturated NaCl (200 mL). The organics are dried over sodium sulfate and the solvent is evaporated to give the title compound. N4-Benzoyl-2'-O-methoxyethyl-5'-O-dimethoxytιϊtyl- 5-methylcytidine
[00124] 2'-O-Methoxyethyl-5'-O-dimethoxytrityl-5-methylcytidine (85 g, 0.134 M) is dissolved in DMF (800 mL) and benzoic anhydride (37.2 g, 0.165 M) is added with stirring. After stirring for 3 hours, TLC showed the reaction to be approximately 95%> complete. The solvent-'is evaporated and the residue azeotroped with MeOH (200 mL). The residue is dissolved in CHC1 (700 mL) and extracted with saturated NaHCO, (2x300 mL) and saturated NaCl (2x300 mL) , dried over MgSO4 and evaporated to give a residue. The residue is chromatographed on a 1.5 kg silica column using EtOAc/hexane (1 :1) containing 0-5% Et NH as the eluting solvent. The pure product fractions are evaporated to give the title compound. N4-Benzoyl-2'-O-methoxyethyl-5'-O-dimethoxytrityl-5-methyIcytidine-3'- amidite [00125] N4-Benzoyl-2'-O-methoxyethyl-5'-O-dimethoxytrityl-5-methylcytidine (74 g, 0.10 M) is dissolved in CH2C12 (1 L) Tetrazole diisopropylamine (7.1 g) and 2-cyanoethoxy-tetra(isopropyl)phosphite (40.5 mL, 0.123 M) are added with stirring, under a nitrogen atmosphere. The resulting mixture is stirred for 20 hours at room temperature (TLC showed the reaction to be 95% complete). The reaction mixture is extracted with saturated NaHCO3 (1x300 mL) and saturated NaCl (3x300 mL). The aqueous washes are back-extracted with CH C12 (300 mL), and the extracts are combined, dried over MgSO4 and concentrated. The residue obtained is chromatographed on a 1.5 kg silica column using EtOAc/hexane (3:1) as the eluting solvent. The pure fractions were combined to give the title compound.
2'-O-(Aminooxyethyl) nucleoside amidites and 2'-O-(dimethylaminooxyethyI) nucleoside amidites
2'-(Dimethylaminooxyethoxy) nucleoside amidites
[00126] 2'-(Dimethylaminooxyethoxy) nucleoside amidites [also known in the art as 2'-O-(dimethylaminooxyethyl) nucleoside amidites] are prepared as described in the following paragraphs. Adenosine, cytidine and guanosine nucleoside amidites are prepared similarly to the thymidine (5-methyluridine) except the exocyclic amines are protected with a benzoyl moiety in the case of adenosine and cytidine and with isobutyryl in the case of guanosine. 5'-O-tert-ButyldiphenyIsilyl -O2 -2'-anhydro-5-methyIuridine [00127] O2 -2'-anhydro-5-methyluridine (Pro. Bio. Sint., Varese, Italy, lOO.Og, 0.4'6 mmol), dimethylaminopyridine (0.66g, 0.013eq, 0.0054mmol) are dissolved in dry pyridine (500 ml) at ambient temperature under an argon atmosphere and with mechanical stirring. tert-Butyldiphenylchlorosilane (125.8g, 1 19.0mL, l .leq, 0.458mmol) is added in one portion. The reaction is stirred for 16 h at ambient temperature. TLC (Rf 0.22, ethyl acetate) indicated a complete reaction. The solution is concentrated under reduced pressure to a thick oil. This is partitioned between dichloromethane (1 L) and saturated sodium bicarbonate (2x1 L) and brine (1 L). The organic layer is dried over sodium sulfate and concentrated under reduced pressure to a thick oil. The oil is dissolved in a 1 :1 mixture of ethyl acetate and ethyl ether (600mL) and the solution is cooled to -10°C. The resulting crystalline product is collected by filtration, washed with ethyl ether (3x200 mL), and dried (40°C, 1mm Hg, 24 h) to a white solid 5'-O-tert-ButyIdiphenylsilyl-2'-O-(2-hydroxyethyl)-5-methyluridine
[00128] In a 2 L stainless steel, unstirred pressure reactor is added borane in tetrahydrofuran (1.0 M, 2.0 eq, 622 mL). In the fume hood and with manual stirring, ethylene glycol (350 mL, excess) is added cautiously at first until the evolution of hydrogen gas subsides. 5'-O-tert-Butyldiphenylsilyl-O2-2'anhydro-5- methyluridine (149 g, 0.3' 1 mol) and sodium bicarbonate (0.074 g, 0.003 eq) are added with manual stirring. The reactor is sealed and heated in an oil bath until an internal temperature of 160°C is reached and then maintained for 16 h (pressure < 100 psig). The reaction vessel is cooled to ambient and opened. TLC (Rf 0.67 for desired product and Rf 0.82 for ara-T side product, ethyl acetate) indicated about 70%> conversion to the product. In order to avoid additional side product formation, the reaction is stopped, concentrated under reduced pressure (10 to 1mm, Hg) in a warm water bath (40-100°C) with the more extreme conditions used to remove the ethylene glycol. [Alternatively, once the low boiling solvent is gone, the remaining solution can be partitioned between ethyl acetate and water. The product will be in the organic phase.] The residue is purified by column chromatography (2kg silica gel, ethyl acetate-hexanes gradient 1 :1 to 4:1). The appropriate fractions are combined, stripped and dried to product as a white crisp foam, contaminated starting material, and pure reusable starting material. 2'-O-([2-phthalimidoxy)ethyl]-5'-t-butyldiphenylsilyl-5-methyluridine [00129] 5'-O-tert-Butyldiphenylsilyl-2'-O-(2-hydroxyethyl)-5- methyluridine (20g, 36.98mmol) is mixed with triphenylphosphine (11.63g, 44.36mmol) and N- hydroxyphthalimide (7.24g, 44.36mmol). It is then dried over P2O under high vacuum for two days at 40°C. The reaction mixture is flushed with argon and dry THF (369.8mL, Aldrich, sure seal bottle) is added to get a clear solution. Diethyl- azodicarboxylate (6.98mL, 44.36mmol) is added dropwise to the reaction mixture. The rate of addition is maintained such that resulting deep red coloration is just discharged before adding the next drop. After the addition is complete, the reaction is stirred for 4 hrs. By that time TLC showed the completion of the reaction (ethylacetate:hexane, 60:40). The solvent is evaporated in vacuum. Residue obtained is placed on a flash column and eluted with ethyl acetate :hexane (60:40), to get 2'-O-([2-phthalimidoxy)ethyl]-5'-t-butyldiphenylsilyl-5-methyluridine as white foam.
5'-O-tert-butyldiphenylsilyl-2'-O-[(2-formadoximinooxy)ethyl]-5- methyluridine [00130] 2'-O-([2-phthalimidoxy)ethyl]-5'-t-butyldiphenylsilyl-5- methyluridine (3.1g, 4.5mmol) is dissolved in dry CH2CI2 (4.5mL) and methylhydrazine (300mL, 4.64mmol) is added dropwise at -10°C to O°C. After 1 h the mixture is filtered, the filtrate is washed with ice cold CH2C12 and the combined organic phase is washed with water, brine and dried over anhydrous Na2SO4. The solution is concentrated to get 2'-O(aminooxyethyl) thymidine, which is then dissolved in MeOH (67.5mL). To this formaldehyde (20% aqueous solution, w/w, 1.1 eq.) is added and the resulting mixture is strirred for 1 h. Solvent is removed under vacuum; residue chromatographed to get 5'-O-tert-butyldiphenylsilyl-2'-O-[(2-formadoximinooxy) ethyl]-5-methyluridine as white foam. 5'-O-tert-ButyldiphenyIsilyl-2'-O-[N,N-dimethylaminooxyethyl]-5- methyluridine
[00131] 5'-O-tert-butyldiphenylsilyl-2'-O-[(2- formadoximinooxy)ethyl]-5- methyluridine (1.77g, 3.12mmol) is dissolved in a solution of IM pyridinium p- toluenesulfonate (PPTS) in dry MeOH (30.6mL). Sodium cyanoborohydride (0.39g, 6.13mmol) is added to this solution at 10°C under inert atmosphere. The reaction mixture is stirred for 10 minutes at 10°C. After that the reaction vessel is removed from the ice bath and stirred at room temperature for 2 h, the reaction monitored by TLC (5% MeOH in CH2C12). Aqueous NaHCO3 solution (5%, lOmL) is added and extracted with ethyl acetate (2x20mL). Ethyl acetate phase is dried over anhydrous Na2SO4, evaporated to dryness. Residue is dissolved in a solution of IM PPTS in MeOH (30.6mL). Formaldehyde (20% w/w, 30mL, 3.37mmol) is added and the reaction mixture is stirred at room temperature for 10 minutes. Reaction mixture cooled to 10°C in an ice bath, sodium cyanoborohydride (0.39g, 6.13mmol) is added, and reaction mixture stirred at 10°C for 10 minutes. After 10 minutes, the reaction mixture is removed from the ice bath and stirred at room temperature for 2 hrs. To the reaction mixture 5% NaHCO3 (25mL) solution is added and extracted with ethyl acetate (2x25mL). Ethyl acetate layer is dried over anhydrous Na2SO4 and evaporated to dryness. The residue obtained is purified by flash column chromatography and eluted with 5% MeOH in CH2C12 to get 5'-O- tertbutyldiphenylsilyl-2'-O-[N,N-dimethylaminooxyethyl]-5- methyluridine as a white foam.
2'-O-(dimethylaminooxyethyl)-5-methyluridine [00132] Triethylamine trihydrofluoride (3.91 mL, 24.0mmol) is dissolved in dry THF and triethylamine (1.67mL, 12mmol, dry, kept over KOH). This mixture of triethylamine-2HF is then added to 5'-O-tert-butyldiphenylsilyl-2'-O-[N,N- dimethylaminooxyethyl]-5-methyluridine (1.40g, 2.4mmol) and stirred at room temperature for 24 hrs. Reaction is monitored by TLC (5% MeOH in CH C12). Solvent is removed under vacuum and the residue placed on a flash column and eluted with 10% MeOH in CH2C12 to get 2'-O-(dimethylaminooxyethyl)-5- methyluridine.
5'-O-DMT-2'-O-(dimethylaminooxyethyl)-5-methyluridine [00133] 2'-O-(dimethylaminooxyethyl)-5-methyluridine (750mg, 2.17mmol) is dried over P2O5 under high vacuum overnight at 40°C. It is then co-evaporated with anhydrous pyridine (20mL). The residue obtained is dissolved in pyridine (1 lmL) under argon atmosphere. 4-dimethylaminopyridine (26.5mg, 2.60mmol), 4,4'- dimethoxytrityl chloride (880mg, 2.60mmol) is added to the mixture and the reaction mixture is stirred at room temperature until all of the starting material disappeared. Pyridine is removed under vacuum and the residue chromatographed and eluted with 10% MeOH in CH2C12 (containing a few drops of pyridine) to get 5'-O-DMT-2'-0(dimethylamino-oxyethyl)-5-methyluridine. 5'-O-DMT-2'-O-(2-N,N-dimethylaminooxyethyl)-5-methyluridine-3'-[(2- cyanoethyl)-N,N- diisopropylphosphoramidite] [00134] 5'-O-DMT-2'-O-(dimethylaminooxyethyl)-5-methyluridine (1.08g, 1.67mmol) is co-evaporated with toluene (20mL). To the residue N,N- diisopropylamine tetrazonide (0.29g, 1.67mmol) is added and dried over P20, under high vacuum overnight at 40°C. Then the reaction mixture is dissolved in anhydrous acetonitrile (8.4mL) and 2-cyanoethyl-N,N,N',N'-tetraisopropylphosphoramidite (2.12mL, 6.08mmol) is added. The reaction mixture is stirred at ambient temperature for 4 hrs under inert atmosphere. The progress of the reaction is monitored by TLC (hexane:ethyl acetate 1 : 1). The solvent is evaporated, then the residue is dissolved in ethyl acetate (70mL) and washed with 5% aqueous NaHC0 (40mL). Ethyl acetate layer is dried over anhydrous Na SO4 and concentrated. Residue obtained is chromatographed (ethyl acetate as eluent) to get 5'-O-DMT-2'- O-(2-N,N-dimethylaminooxyethyl)-5-methyluridine-3'-[(2-cyanoethyl)-N,N- diisopropylphosphoramidite] as a foam. 2'-(Aminooxyethoxy) nucleoside amidites
[00135] 2'-(Aminooxyethoxy) nucleoside amidites [also known in the art as 2'- O-(aminooxyethyl) nucleoside amidites] are prepared as described in the following paragraphs. Adenosine, cytidine and thymidine nucleoside amidites are prepared similarly. N2-isobutyryl-6-O-diphenylcarbamoyl-2'-O-(2-ethylacetyl)-5'-O-(4,4'- dimethoxytrityl)guanosine-3'-[(2-cyanoethyl)-N,N- diisopropylphosphoramidite]
[00136] The 2'-O-aminooxyethyl guanosine analog may be obtained by selective 2'-0-alkylation of diaminopurine riboside. Multigram quantities of diaminopurine riboside may be purchased from Schering AG (Berlin) to provide 2'-O-(2- ethylacetyl) diaminopurine riboside along with a minor amount of the 3'-O-isomer. 2'-O-(2-ethylacetyl) diaminopurine riboside may be resolved and converted to 2'- O-(2ethylacetyl)guanosine by treatment with adenosine deaminase. (McGee, D. P. C, Cook, P. D., Guinosso, C. J., WO 94/02501 Al 940203.) Standard protection procedures should afford 2'-O-(2-ethylacetyl)-5 '-O-(4,4'- dimethoxytrityl)guanosine and 2-N-isobutyryl-6-O-diphenylcarbamoyl-2 '-O-(2- ethylacetyl)-5'-O-(4,4'-dimethoxytrityl)guanosine which may be reduced to provide 2-N-isobutyryl-6-O-diphenylcarbamoyl-2'-O-(2-ethylacetyl)-5'-O-(4,4'- dimethoxytrityl)guanosine. As before the hydroxyl group may be displaced by N- hydroxyphthalimide via a Mitsunobu reaction, and the protected nucleoside may phosphitylated as usual to yield 2-N-isobutyryl-6-O-diphenylcarbamoyl-2'-O-(2- ethylacetyl)-5'-O-(4,4'-dimethoxytrityl)guanosine-3'-[(2-cyanoethyl)-N,N- diisopropylphosphoramiditel. 2'-dimethyIaminoethoxyethoxy (2'-DMAEOE) nucleoside amidites [00137] 2 '-dimethylaminoethoxyethoxy nucleoside amidites (also known in the art as 2'-O-dimethylaminoethoxyethyl, i.e., 2O-CH2-O-CH2-N(CH2)2, or 2'- DMAEOE nucleoside amidites) are prepared as follows. Other nucleoside amidites are prepared similarly. 2'-O-[2(2-N,N-dimethylaminoethoxy)ethyl]-5-methyl uridine [00138] 2[2-(Dimethylamino)ethoxylethanol (Aldrich, 6.66 g, 50 mmol) is slowly added to a solution of borane in tetrahydrofuran (1 M, 10 mL, 10 mmol) with stirring in a 100 mL bomb. Hydrogen gas evolves as the solid dissolves. O -, 2' - anhydro-5-methyluridine (1.2 g, 5 mmol), and sodium bicarbonate (2.5 mg) are added and the bomb is sealed, plaped in an oil bath and heated to 155°C for 26 hours. The bomb is cooled to room temperature and opened. The crude solution is concentrated and the residue partitioned between water (200 mL) and hexanes (200 mL). The excess phenol is extracted into the hexane layer. The aqueous layer is extracted with ethyl acetate (3x200 mL) and the combined organic layers are washed once with water, dried over anhydrous sodium sulfate and concentrated. The residue is columned on silica gel using methanol/methylene chloride 1 :20 (which has 2%> triethylamine) as the eluent. As the column fractions are concentrated a colorless solid forms which is collected to give the title compound as a white solid.
5'-O-dimethoxytrityl-2'-O-[2(2-N,N-dimethylaminoethoxy) ethyl)]-5-methyl uridine
[00139] To 0.5 g (1.3 mmol) of 2'-O-[2(2-N,N-dimethylaminoethoxy)ethyl)l-5- methyl uridine in anhydrous pyridine (8 mL), triethylamine (0.36 mL) and dimethoxytrityl chloride (DMT-Cl, 0.87 g, 2 eq.) are added and stirred for 1 hour. The reaction mixture is poured into water (200 mL) and extracted with CH2C12 (2x200 mL) . The combined CH2CI2 layers are washed with saturated NaHCO3 solution, followed by saturated NaCl solution and dried over anhydrous sodium sulfate. Evaporation of the solvent followed by silica gel chromatography using MeOH: CH2Cl2:Et3N (20: 1 , v/v, with 1% triethylamine) gives the title compound. 5'-O-Dimethoxytrityl-2'-O-[2(2-N,N-dimethylaminoethoxy)ethyl)]-5-nιethyl uridine-3'-O-(cyanoethyl-N,N-diisopropyl)phosphoramidite [00140] Diisopropylaminotetrazolide (0.6 g) and 2-cyanoethoxyN,N-diisopropyl phosphoramidite (1.1 mL, 2 eq.) are added to a solution of 5'-O-dimethoxytrityl-2'- 0-[2(2-N,N-dimethylaminoethoxy)ethyl)]-5-methyluridine (2.17 g, 3 mmol) dissolved in CH2CI2 (20 mL) under an atmosphere of argon. The reaction mixture is stirred overnight and the solvent evaporated. The resulting residue is purified by silica gel flash column chromatography with ethyl acetate as the eluent to give the title compound.
Example 2 Oligonucleotide synthesis
[00141] Unsubstituted and substituted phosphodiester (P=O) oligonucleotides are synthesized on an automated DNA synthesizer (Applied Biosystems model 380B) using standard phosphoramidite chemistry with oxidation by iodine.
[00142] Phosphorothioates (P=S) are synthesized as for the phosphodiester oligonucleotides except the standard oxidation bottle is replaced by 0.2 M solution of 3H-l,2-benzodithiole-3-one 1,1 -dioxide in acetonitrile for the stepwise thiation of the phosphite linkages. The thiation wait step is increased to 68 sec and is followed by the capping step. After cleavage from the CPG column and deblocking in concentrated ammonium hydroxide at 55°C (18 h), the oligonucleotides are purified by precipitating twice with 2.5 volumes of ethanol from a 0.5 M NaCl solution. Phosphinate oligonucleotides are prepared as described in U.S. Patent
5,508,270, herein incoφorated by reference.
[00143] Alkyl phosphonate oligonucleotides are prepared as described in U.S.
Patent 4,469,863, herein incoφorated by reference. [00144] 3 '-Deoxy-3 '-methylene phosphonate oligonucleotides are prepared as described in U.S. Patents 5,610,289 or 5,625,050, herein incoφorated by reference.
[00145] Phosphoramidite oligonucleotides are prepared as described in U.S.
Patent, 5,256,775 or U.S. Patent 5,366,878, herein incoφorated by reference.
[00146] Alkylphosphonothioate oligonucleotides are prepared as described in WO 94/17093 and WO 94/02499 herein incoφorated by reference.
[00147] 3 '-Deoxy-3 '-amino phosphoramidate oligonucleotides are prepared as described in U.S. Patent 5,476,925, herein incoφorated by reference.
[00148] Phosphotriester oligonucleotides are prepared as described in U.S.
Patent 5,023,243, herein incorporated by reference. |00149] Borano phosphate oligonucleotides are prepared as described in U.S.
Patents 5,130,302 and 5,177,198, both herein incoφorated by reference. Example 3
Oligonucleoside Synthesis
[00150] Methylenernethylimino linked oligonucleosides, also identified as MMI linked oligonucleosides, methylenedimethylhydrazo linked oligonucleosides, also identified as MDH linked oligonucleosides, and methylenecarbonylamino linked oligonucleosides, also identified as amide-3 linked oligonucleosides, and methyl eneaminocarbonyl linked oligonucleosides, also identified as amide-4 linked oligonucleosides, as well as mixed backbone compounds having, for instance, alternating MMI and P=O or P=S linkages are prepared as described in U.S. Patents 5,378,825; 5,386,023; 5,489,677; 5,602,240; and 5,610,289, all of which are herein incoφorated by reference.
[00151] Formacetal and thioformacetal linked oligonucleosides are prepared as described in U.S. Patents 5,264,562 and 5,264,564, herein incoφorated by reference. [00152] Ethylene oxide linked oligonucleosides are prepared as described in U.S. Patent 5,223,618, herein incoφorated by reference.
Example 4 PNA Synthesis [00153] Peptide nucleic acids (PNAs) are prepared in accordance with any of the various procedures referred to in Peptide Nucleic Acids (PNA): Synthesis, Properties and Potential Applications, Bioorganic & Medicinal Chemistry, 1996, 4, 523. They may also be prepared in accordance with U.S. Patents 5,539,082; 5,700,922; and 5,719,262, herein incoφorated by reference.
Example 5
Synthesis of Chimeric Oligonucleotides
[00154] Chimeric oligonucleotides, oligonucleosides or mixed oligonucleotides/oligonucleosides of the invention can be of several different types. These include a first type wherein the "gap" segment of linked nucleosides is positioned between 5' and 3' "wing" segments of linked nucleosides and a second "open end" type wherein the "gap" segment is located at either the 3' or the 5' terminus of the oligomeric compound. Oligonucleotides of the first type are also known in the art as "gapmers" or gapped oligonucleotides. Oligonucleotides of the second type are also known in the art as "hemimers" or "wingmers". [2'-O-Me]— [2'-deoxy]--[2'-O-Me] Chimeric Phosphorothioate Oligonucleotides [00155] Chimeric oligonucleotides having 2'-O-alkyl phosphorothioate and 2'- deoxy phosphorothioate oligonucleotide segments are synthesized using an Applied Biosystems automated DNA synthesizer Model 380B, as above. Oligonucleotides are synthesized using the automated synthesizer and 2'-deoxy-5'-dimethoxytrityl- 3'-O-phosphoramidite for the DNA portion and 5'-dimethoxytrityl-2'-O-methyl-3'- O-phosphoramidite for 5' and 3' wings. The standard synthesis cycle is modified by increasing the wait step after the delivery of tetrazole and base to 600 s repeated four times for RNA and twice for 2'-O-methyl. The fully protected oligonucleotide is cleaved from the support and the phosphate group is deprotected in 3:1 ammonia/ethanol at room temperature overnight then lyophilized to dryness. Treatment in methanolic ammonia for 24 hrs at room temperature is then done to deprotect all bases and sample is again lyophilized to dryness. The pellet is resuspended in IM TBAF in THF for 24 hrs at room temperature to deprotect the 2' positions. The reaction is then quenched with IM TEAA and the sample is then reduced to 1/2 volume by rotovac before being desalted on a G25 size exclusion column. The oligo recovered is then analyzed spectrophotometrically for yield and for purity by capillary electrophoresis and by mass spectrometry.
[2'-O-(2-Methoxyethyl)]-[2'-deoxy]--[2'-O-(Methoxyethyl)] Chimeric Phosphorothioate Oligonucleotides
[00156] [2'-O-(2-methoxyethyl)]-[2'-deoxy]—[-2'-O-(methoxyethyl)] chimeric phosphorothioate oligonucleotides are prepared as per the procedure above for the 2'-O-methyl chimeric oligonucleotide, with the substitution of phorothioate oligonucleotides are prepared as per the procedure abo 2'-O-(methoxyethyl) amidites for the 2'-O-methyl amidites.
[2'-O-(2-Methoxyethyl)Phosphodiester]--[2'-deoxy Phosphorothioate]--[2'-O- (2-Methoxyethyl)] Phosphodiester] Chimeric Oligonucleotides [00157] [2'-O-(2-methoxyethyl phosphodiester]~[2'-deoxy phosphorothioate]— [2'-O-(methcixyethyl) phosphodiester] chimeric oligonucleotides are prepared as per the above procedure for the 2'-O-methyl chimeric oligonucleotide with the substitution of 2'-O-(methoxyethyl) amidites for the 2'-O-methyl amidites, oxidization with iodine to generate the phosphodiester internucleotide linkages within the wing portions of the chimeric structures and sulfurization utilizing 3,H- 1 ,2 benzodithiole-3-one 1 ,1 dioxide (Beaucage Reagent) to generate the phosphorothioate internucleotide linkages for the center gap. [00158] Other chimeric oligonucleotides, chimeric oligonucleosides and mixed chimeric oligonucleotides/oligonucleosides are synthesized according to United States patent 5,623,065, herein incoφorated by reference.
Example 6 Oligonucleotide Isolation
[00159] After cleavage from the controlled pore glass column (Applied Biosystems) and deblocking in concentrated ammonium hydroxide at 55°C for 18 hours, the oligonucleotides or oligonucleosides are purified by precipitation twice out of 0.5 M NaCl with 2.5 volumes ethanol. Synthesized oligonucleotides are analyzed by polyacrylamide gel electrophoresis on denaturing gels and judged to be at least 85% full length material. The relative amounts of phosphorothioate and phosphodiester linkages obtained in synthesis are periodically checked by P nuclear magnetic resonance spectroscopy, and for some studies oligonuclectides are purified by HPLC, as described by Chiang et al., J. Biol. Chem. 1991, 266, 18162- 18171.
Example 7
Oligonucleotide Synthesis - 96 Well Plate Format
[00160] Oligonucleotides are synthesized via solid phase P(III) phosphoramidite chemistry on an automated synthesizer capable of assembling 96 sequences simultaneously in a standard 96 well format. Phosphodiester internucleotide linkages are afforded by oxidation with aqueous iodine. Phosphorothioate internucleotide linkages are generated by sulfurization utilizing 3,H-1,2 benzodithiole-3-one 1 ,1 dioxide (Beaucage Reagent) in anhydrous acetonitrile. Standard base -protected beta-cyanoethyldiisopropyl phosphoramidites can be purchased from commercial vendors (e.g. PE-Applied Biosystems, Foster City, CA, or Pharmacia, Piscataway, NJ). Non-standard nucleosides are synthesized as per known literature or patented methods. They are utilized as base protected betacyanoethyldiisopropyl phosphoramidites.
[00161] Oligonucleotides are cleaved from support and deprotected with concentrated NH OH at elevated temperature (55-60°C) for 12-16 hours and the released product then dried in vacuo. The dried product is then re-suspended in sterile water to afford a master plate from which all analytical and test plate samples are then diluted utilizing robotic pipettors.
Example 8 Oligonucleotide Analysis - 96 Well Plate Format
[00162] The concentration of oligonucleotide in each well is assessed by dilution of samples and UV absorption spectroscopy. The full-length integrity of the individual products is evaluated by capillary electrophoresis (CE) in either the 96 well format (Beckman P/ACE™ MDQ) or, for individually prepared samples, on a commercial CE apparatus (e.g., Beckman P/ACE™ 5000, ABI 270). Base and backbone composition is confirmed by mass analysis of the compounds utilizing electrospray-mass spectroscopy. All assay test plates are diluted from the master plate using single and multi-channel robotic pipettors. Plates are judged to be acceptable if at least 85% of the compounds on the plate are at least 85% full length.
Example 9
Cell culture and oligonucleotide treatment
[00163] The effect of antisense compounds on target nucleic acid expression can be tested in any of a variety of cell types provided that the target nucleic acid is present at measurable levels. This can be routinely determined using, for example, PCR or Northern blot analysis. The following 6 cell types are provided for illustrative puφoses, but other cell types can be routinely used, provided that the target is expressed in the cell type chosen. This can be readily determined by methods routine in the art, for example Northern blot analysis, Ribonuclease protection assays, or RT-PCR.
T-24 cells: [00164] The human transitional cell bladder carcinoma cell line T-24 is obtained from the American Type Culture Collection (ATCC) (Manassas, VA). T-24 cells are routinely cultured in complete McCoy's 5A basal media (Gibco/Life Technologies, Gaithersburg, MD) supplemented with 10% fetal calf serum (Gibco/Life Technologies, Gaithersburg, MD), penicillin 100 units per mL, and streptomycin 100 micrograms per mL (Gibco/Life Technologies, Gaithersburg, MD). Cells are routinely passaged by trypsinization and dilution when they reached 90% confluence. Cells are seeded into 96-well plates (Falcon-Primaria #3872) at a density of 7000 cells/well for use in RT-PCR analysis. [00165] For Northern blotting or other analysis, cells may be seeded onto 100 mm or other standard tissue culture plates and treated similarly, using appropriate volumes of medium and oligonucleotide.
A549 cells: [00166] The human lung carcinoma cell line A549 can be obtained from the American Type Culture Collection (ATCC) (Manassas, VA). A549 cells are routinely cultured in DMEM basal media (Gibco/Life Technologies, Gaithersburg, MD) supplemented with 10% fetal calf serum (Gibco/Life Technologies, Gaithersburg, MD), penicillin 100 units per mL, and streptomycin 100 micrograms per mL (Gibco/Life Technologies, Gaithersburg, MD). Cells are routinely passaged by trypsinization and dilution when they reached 90% confluence.
NHDF cells:
[00167] Human neonatal dermal fibroblast (NHDF) can be obtained from the Clonetics Coφoration (Walkersville MD). NHDFs are routinely maintained in Fibroblast Growth Medium (Clonetics Coφoration, Walkersville MD) supplemented as recommended by the supplier. Cells are maintained for up to 10 passages as recommended by the supplier.
HEK cells:
|00168] Human embryonic keratinocytes (HEK) can be obtained from the Clonetics Corporation (Walkersville MD). HEKs are routinely maintained in Keratinocyte Growth Medium (Clonetics Corporation, Walkersville MD) formulated as recommended by the supplier. Cells are routinely maintained for up to 10 passages as recommended by the supplier.
MCF-7 cells: [00169] The human breast carcinoma cell line MCF-7 is obtained from the American Type Culure Collection (Manassas, VA). MCF-7 cells are routinely cultured in DMEM low glucose (Gibco/Life Technologies, Gaithersburg, MD) supplemented with 10%> fetal calf serum (Gibco/Life Technologies, Gaithersburg, MD). Cells are routinely passaged by trypsinization and dilution when they reached 90% confluence. Cells are seeded into 96-well plates (Falcon-Primaria #3872) at a density of 7000 cells/well for use in RT-PCR analysis.
[00170] For Northern blotting or other analyses, cells may be seeded onto 100 mm or other standard tissue culture plates and treated similarly, using appropriate volumes of medium and oligonucleotide.
LA4 cells:
[00171] The mouse lung epithelial cell line LA4 is obtained from the American Type Culure Collection (Manassas, VA). LA4 cells are routinely cultured in F12K medium (Gibco/Life Technologies, Gaithersburg, MD) supplemented with 15% fetal calf serum (Gibco/Life Technologies, Gaithersburg, MD). Cells are routinely passaged by trypsinization and dilution when they reached 90% confluence. Cells are seeded into 96-well plates (Falcon-Primaria #3872) at a density of 3000-6000 cells/ well for use in RT-PCR analysis. [00172] For Northern blotting or other analyses, cells may be seeded onto 100 mm or other standard tissue culture plates and treated similarly, using appropriate volumes of medium and oligonucleotide. [00173] Treatment with antisense compounds: [00174] When cells reached 80% confluency, they are treated with oligonucleotide. For cells grown in 96-well plates, wells are washed once with 200 μL OPTl-MEM,m-] reduced-serum medium (Gibco BRL) and then treated with 130 μL of OPTI-MEMTM™-l containing 3.75 μg/mL LIPOFECT1N™ (Gibco BRL) and the desired concentration of oligonucleotide. After 4-7 hours of treatment, the medium is replaced with fresh medium. Cells are harvested 16-24 hours after oligonucleotide treatment.
[00175] The concentration of oligonucleotide used varies from cell line to cell line. To determine the optimal oligonucleotide concentration for a particular cell line, the cells are treated with a positive control oligonucleotide at a range of concentrations. If 80% inhibition is not achieved, the lowest concentration of positive control oligonucleotide that results in 60% inhibition of H-ras or c-raf mRNA is then utilized as the oligonucleotide screening concentration in subsequent experiments for that cell line. If 60% inhibition is not achieved, that particular cell line is deemed as unsuitable for oligonucleotide transfection experiments.
Example 10
Analysis of oligonucleotide inhibition of GR expression
[00176] Antisense modulation of GR expression can be assayed in a variety of ways known in the art. For example, GR mRNA levels can be quantitated by, e.g., Northern blot analysis, competitive polymerase chain reaction (PCR), or real-time PCR (RT-PCR). Real-time quantitative PCR is presently preferred. RNA analysis can be performed on total cellular RNA or poly(A)+ mRNA. Methods of RNA isolation are taught in, for example, Ausubel, F.M. et al., Current Protocols in Molecular Biology, Volume 1, pp. 4.1.1-4.2.9 and 4.5.1-4.5.3, John Wiley & Sons, Inc., 1993. Northern blot analysis is routine in the art and is taught in, for example, Ausubel, F.M. et al., Current Protocols in Molecular Biology, Volume 1, pp. 4.2.1- 4.2.9, John Wiley & Sons, Inc., 1996. Real-time quantitative (PCR) can be conveniently accomplished using the commercially available ABI PRISM™ 7700 Sequence Detection System, available from PE-Applied Biosystems, Foster City, CA and used according to manufacturer's instructions. Prior to quantitative PCR analysis, primer-probe sets specific to the target gene being measured are evaluated for their ability to be "multiplexed" with a GAPDH amplification reaction. In multiplexing, both the target gene and the internal standard gene GAPDH are amplified concurrently in a single sample. In this analysis, mRNA isolated from untreated cells is serially diluted. Each dilution is amplified in the presence of primer-probe sets specific for GAPDH only, target gene only ("single-plexing"), or both (multiplexing). Following PCR amplification, standard curves of GAPDH and target mRNA signal as a function of dilution are generated from both the single- plexed and multiplexed samples. If both the slope and correlation coefficient of the GAPDH and target signals generated from the multiplexed samples fall within 10% of their corresponding values generated from the single-plexed samples, the primer- probe set specific for that target is deemed as multiplexable. Other methods of PCR are also known in the art.
[00177] Protein levels of GR can be quantitated in a variety of ways well known in the art, such as immunoprecipitation, Western blot analysis (immunoblotting), ELISA or fluorescence-activated cell sorting (FACS). Antibodies directed to GR can be identified and obtained from a variety of sources, such as the MSRS catalog of antibodies (Aerie Corporation, Birmingham, MI), or can be prepared via conventional antibody generation methods. Methods for preparation of polyclonal antisera are taught in, for example, Ausubel, F.M. et al., Current Protocols in Molecular Biology, Volume 2, pp. 11.12.1-11.12.9, John Wiley & Sons, Inc., 1997. Preparation of monoclonal antibodies is taught in, for example, Ausubel, F.M. et al., Current Protocols in Molecular Biology, Volume 2, pp. 11.4.1 -11.11.5, John Wiley Sons, Inc., 1997.
[00178] Immunoprecipitation methods are standard in the art and can be found at, for example, Ausubel, F.M. et al., Current Protocols in Molecular Biology, Volume 2, pp. 10.16.1 10.16.11, John Wiley & Sons, Inc., 1998. Western blot (immunoblot) analysis is standard in the art and can be found at, for example, Ausubel, F.M. et al., Current Protocols in Molecular Biology, Volume 2, pp. 10.8.1-10.8.21 , John Wiley Sons, Inc., 1997. Enzyme-linked immunosorbent assays (ELISA) are standard in the art and can be found at, for example, Ausubel, F.M. et al., Current Protocols in Molecular Biology, Volume 2, pp. 1 1.2.1-11.2.22, John Wiley & Sons, Inc., 1991.
Example 11
Poly(A)+ mRNA isolation [00179] Poly(A)+ mRNA is isolated according to Miura et al., Clin. Chem., 1996, 42, 1758-1764. Other methods for poly(A)+ mRNA isolation are taught in, for example, Ausubel, F.M. et al., Current Protocols in Molecular Biology, Volume 1 , pp. 4.5.1-4.5.3, John Wiley & Sons, Inc., 1993. Briefly, for cells grown on 96- well plates, growth medium is removed from the cells and each well is washed with 200 μL cold PBS. 60μL lysis buffer (10 mM Tris-HCl, pH 7.6, 1 mM EDTA, 0.5 M NaCl, 0.5%) NP-40, 20 mM vanadyl-ribonucleoside complex) is added to each well, the plate is gently agitated and then incubated at room temperature for five minutes. 55 μL of lysate is transferred to Oligo d(T) coated 96-well plates (AGCT Inc., Irvine CA). Plates are incubated for 60 minutes at room temperature, washed 3 times with 200 μL of wash buffer (10 mM Tris-HCl pH 7.6, 1 mM EDTA, 0.3 M NaCl). After the final wash, the plate is blotted on paper towels to remove excess wash buffer and then air-dried for 5 minutes. 60 pL of elution buffer (5 mM Tris- HCl pH 7.6), preheated to 70°C is added to each well, the plate is incubated on a 90°C hot plate for 5 minutes, and the eluate is then transferred to a fresh 96-well plate.
[00180] Cells grown on 100 mm or other standard plates may be treated similarly, using appropriate volumes of all solutions.
Example 12
Total RNA Isolation
[00181] Total mRNA is isolated using an RNEASY 96™ kit and buffers purchased from Qiagen Inc. (Valencia CA) following the manufacturer's recommended procedures. Briefly, for cells grown on 96-well plates, growth medium is removed from the cells and each well is washed with 200 μL cold PBS. 100 μL Buffer RLT is added to each well and the plate vigorously agitated for 20 seconds. 100 μL of 70%> ethanol is then added to each well and the contents mixed by pipetting three times up and down. The samples are then transferred to the RNEASY 96™ well plate attached to a QIA VAC™ manifold fitted with a waste collection tray and attached to a vacuum source. Vacuum is applied for 15 seconds. 1 mL of Buffer RW1 is added to each well of the RNEASY 96™ plate and the vacuum again applied for 15 seconds. 1 mL of Buffer RPE is then added to each well of the RNEASY 96 plate and the vacuum applied for a period of 15 seconds. The Buffer RPE wash is then repeated and the vacuum is applied for an additional 10 minutes. The plate is then removed from the QIAVAC™ manifold and blotted dry on paper towels. The plate is then re-attached to the QIAVAC™ manifold fitted with a collection tube rack containing 1.2 mL collection tubes. RNA is then eluted by pipetting 60μL water into each well, incubating I minute, and then applying the vacuum for 30 seconds. The elution step is repeated with an additional 60μL water. [00182] The repetitive pipetting and elution steps may be automated using a QIAGEN Bio-Robot 9604 (Qiagen, Inc., Valencia CA). Essentially, after lysing of the cells on the culture plate, the plate is transferred to the robot deck where the pipetting, DNase treatment and elution steps are carried out.
Example 13
Real-time Quantitative PCR Analysis of GR mRNA Levels [00183] Quantitation of GR mRNA levels is determined by real-time quantitative PCR using the ABI PRISM™ 7700 Sequence Detection System (PE-Applied Biosystems, Foster City, CA) according to manufacturer's instructions. This is a closed-tube, non-gel-based, fluorescence detection system which allows high- throughput quantitation of polymerase chain reaction (PCR) products in real-time. As opposed to standard PCR, in which amplification products are quantitated after the PCR is completed, products in real-time quantitative PCR are quantitated as they accumulate. This is accomplished by including in the PCR reaction an oligonucleotide probe that anneals specifically between the forward and reverse PCR primers, and contains two fluorescent dyes. A reporter dye (e.g., JOE, FAM, or VIC, obtained from either Operon Technologies Inc., Alameda, CA or PE- Applied Biosystems, Foster City, CA) is attached to the 5' end of the probe and a quencher dye (e.g., TAMRA, obtained from either Operon Technologies Inc., Alameda, CA or PE-Applied Biosystems, Foster City, CA) is attached to the 3' end of the probe. When the probe and dyes are intact, reporter dye emission is quenched by the proximity of the 3' quencher dye. During amplification, annealing of the probe to the target sequence creates a substrate that can be cleaved by the 5'- exonuclease activity of Taq polymerase. During the extension phase of the PCR amplification cycle, cleavage of the probe by Taq polymerase releases the reporter dye from the remainder of the probe (and hence from the quencher moiety) and a sequence-specific fluorescent signal is generated. With each cycle, additional reporter dye molecules are cleaved from their respective probes, and the fluorescence intensity is monitored at regular intervals by laser optics built into the
ABI PRISM 7700 Sequence Detection System. In each assay, a series of parallel reactions containing serial dilutions of mRNA from untreated control samples generates a standard curve that is used to quantitate the percent inhibition after antisense oligonucleotide treatment of test samples.
[00184] PCR reagents can be obtained from PE-Applied Biosystems, Foster City, CA. RT-PCR reactions are carried out by adding 25μL PCR cocktail (lx
TAQMAN™ buffer A, 5.5 MM MgCl2, 300 μM each of dATP, dCTP and dGTP, 600 μM of dUTP, 100 nM each of forward primer, reverse primer, and probe, 20
Units RNAse inhibitor, 1.25 Units AMPLITAQ GOLD , and 12.5 Units MuLV reverse transcriptase) to 96 well plates containing 25 μL poly(A) mRNA solution. The RT reaction is carried out by incubation for 30 minutes at 48°C. Following a 10 minute incubation at 95°C to activate the AMPLITAQ GOLD , 40 cycles of a two- step PCR protocol are carried out: 95°C for 15 seconds (denaturation) followed by 60°C for 1.5 minutes (annealing/extension).
[00185] Probes and primers to human GR were designed to hybridize to a human GR sequence shown in Figure 1 , using published sequence information (GenBank accession number NM_000176, incoφorated herein). For human GR the PCR primers were: forward primer: AGGTTGTGCAAATTAACAGTCCTAACT SEQ ID NO : 4774 reverse primer: TAGTCTTTTGCAACCATCATCCA SEQ ID NO : 4775 and the PCR probe is: FA -AGCACCTAGTCCAGTGACCTGCTGGGTAAA-TAMPA SEQ ID
NO : 4776 where FAM (PE-Applied Biosystems, Foster City, CA) is the fluorescent reporter dye) and TAMRA (PE-Applied Biosystems, Foster City, CA) is the quencher dye. For human cyclophilin the PCR primers were: forward primer: CCCACCGTGTTCTTCGACAT SEQ ID NO : 4777 reverse primer: TTTCTGCTGTCTTTGGGACCTT SEQ ID NO : 4778 and the PCR probe is: JOE-CGCGTCTCCTTTGAGCTGTTTGCA-TAMRA SEQ ID NO: 4779 where
JOE (PE-Applied Biosystems, Foster City, CA) is the fluorescent reporter dye) and TAMRA (PE-Applied Biosystems, Foster City, CA) is the quencher dye. |00186] Probes and primers to mouse GR were designed to hybridize to a mouse GR sequence shown in Figure 2, using published sequence information (GenBank NM_008173 incoφorated herein). For mouse GR the PCR primers were:
[00187] forward primer: CGGGACCACCTCCCAAAC SEQ ID NO : 4780 reverse primer: GCACCCCATAATGGCATACC SEQ ID NO : 4781 and the PCR probe is: FAM-TGCCTGGTGTGCTCCGATGAAGC- TAMPA SEQ ID NO : 4782 where FAM
(PE-Applied Biosystems, Foster City, CA) is the fluorescent reporter dye) and TAMRA (PEApplied Biosystems, Foster City, CA) is the quencher dye. For mouse cyclophilin the PCR primers were: forward primer: AGAGAAATTTGAGGATGAGAACTTCAT SEQ ID NO : 4783 reverse primer: TTGTGTTTGGTCCAGCATTTG SEQ ID NO : 4784 and the PCR probe is: JOE- AAGCATACAGGTCCTGGCATCTTGTCCAT- TAMRA SEQ ID NO : 4785 where JOE (PE-Applied Biosystems, Foster City, CA) is the fluorescent reporter dye) and TAMRA (PEApplied Biosystems, Foster City, CA) is the quencher dye.
Example 14
Antisense inhibition of human GR expression by chimeric phosphorothioate oligonucleotides having 2'---VIOE wings and a deoxy gap [00188] In accordance with the present invention, a series of oligonucleotides are designed to target different regions of the human GR RNA, using published sequences (GenBank accession number NM_000176.1). The oligonucleotides are shown in Table 1. "Target site" indicates the first (5 '-most) nucleotide number on the particular target sequence to which the oligonucleotide binds. The indicated parameters for each oligo were predicted using RNAstructure 3.7 by David H.
Mathews, Michael Zuker and Douglas H. Turner. RNAstructure is available on the Turner Lab Homepage at http://rna.chem.rochester.edu. The other modern implementation of the algorithm is mfold, available on the World Wide Web at Michael Zuker's homepage, http://bioinfo.math.φi.edu/~zukerm/. The RNA folding algorithm is based on the following research:D.H. Mathews, et al., Journal of Molecular Biology, 288, 91 1 -940, (1999). OligoWalk is based on the following research: D.H. Mathews, et al., RNA, 5, 1458-1469 (1999). The Dynalign algorithm is based on the following research: D.H. Mathews and D.H. Turner, Journal of Molecular Biology 317(2), 191-203 (2002). The mix&match algorithm is based on the following research: D. H. Mathews, et al., RNA, 3, 1 -16, (1997). The DNA parameters for folding are derived from:J. SantaLucia, Jr., Proc. Natl. Acad. Sci. USA 95, 1460-1465, (1998). DNA oligomers in oligowalk use the DNA/RNA duplex parameters of: N. Sugimoto, et al. Biochemistry, 34, 1 121 1-11216, (1995).
[00189] The oligos are sorted by the predicted free energy of the oligo interupting the predicted secondary structure of the messenger RNA. All compounds in Table 1 are chimeric oligonucleotides ("gapmers") 20 nucleotides in length, composed of a central "gap" region consisting often 2'deoxynucleotides, which is flanked on both sides (5' and 3' directions) by four-nucleotide "wings". The wings are composed of 2 '-methoxy ethyl (2'-MOE) nucleotides. The internucleoside (backbone) linkages are phosphorothioate (P=S) throughout the oligonucleotide. Cytidine residues in the 2'-MOE wings are 5-methylcytidines. All cytidine residues are 5-methylcytidines.
TABLE 1 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecu Site oligo binding formation Duplex structure oligo oligo
AGGGTGGTCAGAATGGGAGG
4358 SEQ ID NO:l -38 -41.9 93 -3.9 0 -5.5
GGGTGGTCAGAATGGGAGGC
4357 SEQ ID NO: 2 -37.6 -43.2 94.1 -5.6 0 -5.5
TGTGAGACTCCTGTAGTGGC
1639 SEQ ID NO: 3 -37.2 -40 90.8 -2.1 -0.4 -6
TTGTGAGACTCCTGTAGTGG
1640 SEQ ID NO: 4 -36.8 -37.5 87.9 -0.5 0 -5.7
CTTGTGAGACTCCTGTAGTG
1641 SEQ ID NO: 5 -35.6 -36.3 86.2 -0.5 0 -5.8
AAGGGTGGTCAGAATGGGAG
4359 SEQ ID NO: 6 -35.6 -39.5 90.2 -3.9 0 -5.3
TCTTGTGAGACTCCTGTAGT
1642 SEQ ID NO: 7 -35.3 -36.6 86.4 -0.5 0 -8.4
TGAGACTCCTGTAGTGGCCT
1637 SEQ ID NO: 8 -35 -41.1 92.5 -5.4 -0.4 -8.3
TTGCTCCAGGAAAGCTTGCC
1041 SEQ ID NO: 9 -34.9 -39.1 90.6 -2.8 -1.2 -10.1
TGCTCCAGGAAAGCTTGCCT
1040 SEQ ID NO: 10 -34.8 -40.3 92.2 -2.8 -1 -13.5
GAGACTCCTGTAGTGGCCTG
1636 SEQ ID NO: 11 -34.8 -41.1 92 -5.4 -0.4 -9.2
GTGAGACTCCTGTAGTGGCC
1638 SEQ ID NO: 12 -34.5 -41.2 92.1 -6 -0.4 -6.6
AGACTCCTGTAGTGGCCTGC
1635 SEQ ID NO: 13 -34.1 -42.1 93.8 -6.9 -1 -8.3
TTCTTGTGAGACTCCTGTAG
1643 SEQ ID NO: 14 -33.8 -35.3 84.8 -0.5 0 -9.3
GGTGGTCAGAATGGGAGGCA
4356 SEQ ID NO: 15 -33.4 -42 92.9 -7.7 -0.8 -5.5
TGCTTGGAGTCTGATTGAGA
286 SEQ ID NO: 16 -33.3 -36.5 85.3 -2.4 -0.2 -9.1
ACCCAGCAGGTCACTGGACT
3318 SEQ ID NO: 17 -33.2 -42.9 94.8 -5.6 -4.1 -13.7
GGGTCAGAGCCTCAGCAACC
4309 SEQ ID NO: 18 -33.1 -43.1 94.2 -7.9 -2.1 -10.6
AGAAGGGTGGTCAGAATGGG
4361 SEQ ID NO: 19 -33 -39.5 90.2 -6.5 0 -5.5
GCTGATTTGGCCCTGCTGTG
446 SEQ ID NO: 20 -32.7 -40.3 91.4 -6 -1.4 -10.1
TCGCTGCTTGGAGTCTGATT
290 SEQ ID NO: 21 -32.6 -37.8 87.5 -4.5 0 -9.1
CTTCGCTGCTTGGAGTCTGA
292 SEQ ID NO: 22 -32.6 -38.8 89 -4.6 -1.5 -8.9
TCTTCGCTGCTTGGAGTCTG
293 SEQ ID NO: 23 -32.6 -38.8 89 -4.6 -1.6 -9.5
TCACTTGGGGCAGTGTTACA
942 SEQ ID NO: 24 -32.6 -38.2 90 -3.8 -1.8 -10.7 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TTTGCTCCAGGAAAGCTTGC
1042 SEQ ID NO: 25 -32.6 -36.7 87.6 -2.8 -1.2 -8.8
TCTCACTGGGTCAGAGCCTC
4316 SEQ ID NO: 26 -32.6 -42.4 92.6 -7.5 -2.3 -11.1
CAGCAGGTCACTGGACTAGG
3315 SEQ ID NO: 27 -32.5 -40.8 92.1 -5.6 -2.7 -10.4
TGGAGTCTGATTGAGAAGCG
282 SEQ ID NO: 28 -32.4 -36.8 85.7 -4.4 0.1 -6.1
TACCCAGCAGGTCACTGGAC
3319 SEQ ID NO: 29 -32.4 -42.1 93.9 -5.6 -4.1 -13.7
GCTGCTTGGAGTCTGATTGA
288 SEQ ID NO: 30 -32.3 -37.5 86.8 -4.5 0 -9
CGCTGCTTGGAGTCTGATTG
289 SEQ ID NO: 31 -32.3 -37.5 87.2 -4.5 0 -9.1
TTTCTTGTGAGACTCCTGTA
1644 SEQ ID NO: 32 -32.3 -34.1 83.4 -0.5 -0.7 -9.3
AGCAGGTCACTGGACTAGGT
3314 SEQ ID NO: 33 -32.3 -40.9 92.7 -5.6 -3 -10.7
CCCAGCAGGTCACTGGACTA
3317 SEQ ID NO: 34 -32.3 -42 93.8 -5.6 -4.1 -13.7
GGTCAGAGCCTCAGCAACCT
4308 SEQ ID NO: 35 -32.3 -41.9 93 -7.9 -1.6 -10.5
TGAATCGTCTTCTCCCGCCA
815 SEQ ID NO: 36 -32.2 -39.5 89.5 -7.3 0 -2.3
GCTCCAGGAAAGCTTGCCTG
1039 SEQ ID NO: 37 -32.2 -40.3 91.7 -2.8 -3.5 -18.7
AGACATTTTCGATAGCGGCA
1558 SEQ ID NO: 38 -32.2 -34.8 84 -2.1 0 -7.5
GCTTGGAGTCTGATTGAGAA
285 SEQ ID NO: 39 -32.1 -35.3 83.8 -2.4 -0.2 -9.1
CTCACTGGGTCAGAGCCTCA
4315 SEQ ID NO: 40 -32.1 -42.1 92.7 -7.5 -2.5 -10.6
GGGTGCAGAGTTCGATGAAA
978 SEQ ID NO: 41 -32 -36.6 86 -3.6 -0.9 -4.9
GGGGTGCAGAGTTCGATGAA
979 SEQ ID NO: 42 -31.6 -39 88.9 -6.4 -0.9 -4.8
TTCACTTGGGGCAGTGTTAC
943 SEQ ID NO: 43 -31.4 -37 88.5 -3.8 -1.8 -8.2
CCAGCAGGTCACTGGACTAG
3316 SEQ ID NO: 44 -31.4 -40.8 92.1 -5.6 -3.8 -13
CTGCTTGGAGTCTGATTGAG
287 SEQ ID NO: 45 -31.3 -36.2 85.1 -4.2 0 -9.1
AGGGGTGCAGAGTTCGATGA
980 SEQ ID NO: 46 -31.3 -40.2 90.5 -7.9 -0.9 -4.6
CATGCTGGGGCTTGAATAGC
1307 SEQ ID NO: 7 -31.3 -38.4 89.1 -5.4 -1.7 -9.6
GTTTACCCAGCAGGTCACTG
3322 SEQ ID NO: 48 -31.3 -38.2 89.7 -5.6 -1.2 -6.4
ATCTCACTGGGTCAGAGCCT
4317 SEQ ID NO: 49 -31.3 -41.1 91.4 -7.5 -2.2 -11.9
TTCTCTGGAACACTGGTCGA
526 SEQ ID NO: 50 -31.2 -37.9 87.7 -6 -0.4 -6.3 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
GACATTTTCGATAGCGGCAT 1557 SEQ ID NO: 51 -31.2 -33.8 82.2 -2.1 0 -7.5 TTACCCAGCAGGTCACTGGA
3320 SEQ ID NO: 52 -31.1 -40.8 92.8 -5.6 -4.1 -13.7 CCATCAGCATTTCTTTGACC
4172 SEQ ID NO: 53 -31.1 -33.7 82.2 -2.6 0 -2.7 TCAGAGCCTCAGCAACCTTC
4306 SEQ ID NO: 54 -31.1 -39.7 90.1 -7.9 -0.5 -7.9 TTCGCTGCTTGGAGTCTGAT
291 SEQ ID NO: 55 -31 -37.8 87.5 -6 0 -9.4 TTTACCCAGCAGGTCACTGG
3321 SEQ ID NO: 56 -31 -39.3 91.4 -5.6 -2.7 -11.1 AGCCGCTCGCCCGCCACCGT
88 SEQ ID"N0:57 -30.9 -48.7 102.1 -17.1 -0.5 -5.7 GGCTGATTTGGCCCTGCTGT
447 SEQ ID NO: 58 -30.9 -41.5 93.2 -5.8 -4.7 -16.7 GGTCTCATGCTGGGGCTTGA
1312 SEQ ID NO: 59 -30.9 -42 92.8 -5.4 -5.5 -18.7
GGTTCTCTGGAACACTGGTC 528 SEQ ID NO: 60 -30.8 -38.6 88.6 -4.4 -3.4 -12
CACTTGGGGCAGTGTTACAT 941 SEQ ID NO: 61 -30.8 -36.9 88.2 -3.8 -1.7 -12.6
GGTGCAGAGTTCGATGAAAT 977 SEQ ID NO: 62 -30.8 -34.4 82.6 -3.6 0 -4.9
CTCCACCCCAGAGCAAATGC 1945 SEQ ID NO: 63 -30.8 -40.5 91.4 -9.7 0 -4.5
ACTGGGTCAGAGCCTCAGCA 4312 SEQ ID NO: 64 -30.8 -43.1 94.5 -7.3 -5 -13.2
GGAATGAGAAGGGTGGTCAG
4367 SEQ ID NO: 65 -30.8 -38.6 88.4 -7.8 0 -5.5 TGGAATGAGAAGGGTGGTCA
4368 SEQ ID NO: 66 -30.8 -38.6 88.7 -7.8 0 -3.6 TCCACAGTTTACCCAGCAGG
3328 SEQ ID NO: 67 -30.7 -39.3 91.4 -8.6 0 -4.3 GTCAGAGCCTCAGCAACCTT
4307 SEQ ID NO: 68 -30.7 -39.5 90.3 -7.9 -0.7 -7.9 GAAGGGTGGTCAGAATGGGA
4360 SEQ ID NO: 69 -30.7 -39.8 90.1 -9.1 0 -5.5 AGGCTGATTTGGCCCTGCTG
448 SEQ ID NO: 70 -30.6 -41.4 93.1 -5.8 -4.9 -17.1 GCAGGTCACTGGACTAGGTG
3313 SEQ ID NO: 71 -30.6 -40.9 92.2 -7.3 -3 -10.7
CAGAGCCTCAGCAACCTTCA 4305 SEQ ID NO: 72 -30.6 -39.4 90.2 -7.9 -0.7 -7.9
ATTTGCTCCAGGAAAGCTTG 1043 SEQ ID NO: 73 -30.5 -34.4 84.2 -2.8 -1 -8.8
AATCTCACTGGGTCAGAGCC 4318 SEQ ID NO: 74 -30.5 -39.9 89.8 -7.5 -0.4 -11.9
CACTGTTGGAATGAGAAGGG 4374 SEQ ID NO: 75 -30.5 -35.9 85.5 -5.4 0 -1
AAGACATTTTCGATAGCGGC 1559 SEQ ID NO: 76 -30.4 -33.6 82.5 -3.2 0 -4.5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecul Site oligo binding formation Duplex structure oligo oligo
GGAGTCTGATTGAGAAGCGA
281 SEQ ID NO: 77 -30.3 -37.1 85.7 -6.3 -0.1 -6.1
TTGGAGTCTGATTGAGAAGC
283 SEQ ID NO: 78 -30.3 -35.3 83.8 -4.4 -0.1 -8.3
TCTGGAACACTGGTCGACCT
523 SEQ ID NO: 79 -30.3 -40.1 90.8 -5.6 -0.8 -16.5
TGGTCTCATGCTGGGGCTTG
1313 SEQ ID NO: 80 -30.3 -41.7 92.9 -5.4 -5.7 -19.8
ATGAATCGTCTTCTCCCGCC
816 SEQ ID NO: 81 -30.2 -38.5 87.6 -8.3 0 -3.8
ATGCTGGGGCTTGAATAGCC
1306 SEQ ID NO: 82 -30.2 -39.6 90.8 -5.4 -4 -14.7
TCCATCAGCATTTCTTTGAC
4173 SEQ ID NO: 83 -30.2 -32.8 80.5 -2.6 0 -2.7
TTTCACTTGGGGCAGTGTTA
944 SEQ ID NO: 84 -30.1 -35.7 87.1 -3.8 -1.8 -8.2
TGCAACCATCATCCACAGTT
3339 SEQ ID NO: 85 -30 -36 85.4 -6 0 -4.1
TTGCAACCATCATCCACAGT
3340 SEQ ID NO: 86 -30 -36 85.4 -6 0 -5.3
GTGGTCAGAATGGGAGGCAG
4355 SEQ ID NO: 87 -30 -40.8 91.2 -9.9 -0.8 -5.5
GTTCTCTGGAACACTGGTCG
527 SEQ ID NO: 88 -29.9 -37.7 87.5 -4.4 -3.4 -12.3
GTGCAGAGTTCGATGAAATC
976 SEQ ID NO: 89 -29.9 -33.5 80.9 -3.6 0 -5.2
ATATTTGCTCCAGGAAAGCT
1045 SEQ ID NO: 90 -29.9 -33.8 83.3 -2.8 -1 -8.8
GAATGAGAAGGGTGGTCAGA
4366 SEQ ID NO: 91 -29.9 -37.7 86.9 -7.8 0 -5.5
TCTCCACCCCAGAGCAAATG
1946 SEQ ID NO: 92 -29.8 -39.5 90 -9.2 -0.1 -3
TTTTCACTTGGGGCAGTGTT
945 SEQ ID NO: 93 -29.7 -35.3 86.4 -3.8 -1.8 -6.4
TATTTGCTCCAGGAAAGCTT
1044 SEQ ID NO: 94 -29.7 -33.6 83.5 -2.8 -1 -8.8
TATATTTGCTCCAGGAAAGC
1046 SEQ ID NO: 95 -29.7 -33 82.3 -2.8 0 -7.6
TCTCATGCTGGGGCTTGAAT
1310 SEQ ID NO: 96 -29.7 -38.5 88.4 -5.4 -3.4 -13.6
ACTAGGTGCTCTATACCAGT
3301 SEQ ID NO: 97 -29.7 -36.8 88.1 -3.5 -3.6 -10.2
CCACAGTTTACCCAGCAGGT
3327 SEQ ID NO: 98 -29.7 -39.1 91.5 -8.6 -0.6 -5.6
ACTGTTGGAATGAGAAGGGT
4373 SEQ ID NO: 99 -29.7 -36 85.8 -5.4 -0.7 -3.6
CTGATTTGGCCCTGCTGTGG
445 SEQ ID NO: 100 -29.6 -40.2 91.5 -10.1 -0.2 -7.5
ACTTGGGGCAGTGTTACATT
940 SEQ ID NO: 101 -29.6 -35.7 86.9 -3.8 -1.4 -12.6
TGGGCACTGGTGGTTTAGGT
2767 SEQ ID NO: 102 -29.6 -40.3 93.3 -8.6 -1 -12.2 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
AGTTTACCCAGCAGGTCACT
3323 SEQ ID NO: 103 -29.6 -38.2 90.1 -7 -1.5 -7.3
GAGAAGGGTGGTCAGAATGG
4362 SEQ ID NO: 104 -29.6 -38.6 88.4 -9 0 -5.5
TTGGAATGAGAAGGGTGGTC
4369 SEQ ID NO: 105 -29.6 -37.4 87.2 -7.8 0 0
TCACTGTTGGAATGAGAAGG
4375 SEQ ID NO: 106 -29.6 -35 83.9 -5.4 0 -3.2
CCACCGTTGGTGCCAGTCTG
649 SEQ ID NO: 107 -29.5 -41.7 93.5 -9.3 -2.6 -13.6
TGTTGAGAAAGGGATGCTGT
1144 SEQ ID NO: 108 -29.5 -36 85.7 -6.5 0 -4.2
TGCTGGGGCTTGAATAGCCA
1305 SEQ ID NO: 109 -29.5 -40.6 92.7 -5.4 -5.7 -14.7
GACTAGGTGCTCTATACCAG
3302 SEQ ID NO: 110 -29.5 -37 87.6 -4.8 -2.7 -8.2
AAGGCTGATTTGGCCCTGCT
449 SEQ ID NO: 111 -29.4 -40.2 92.1 -5.8 -4.9 -17.1
TCAGAGCACACCAGGCAGAG
1390 SEQ ID NO: 112 -29.3 -41.9 92.7 -11.8 -0.6 -5.2
CCGCCGCAGCCGAGATAAAC
59 SEQ ID NO: 113 -29.2 -40.7 92.1 -11 0 -7.5
GAGCCGCTCGCCCGCCACCG
89 SEQ ID NO: 114 -29.2 -48.9 101.1 -16.4 -0.5 -14.7
CTGGAACACTGGTCGACCTA
522 SEQ ID NO: 115 -29.2 -39 90 -5.6 -0.8 -16.5
CAGAGTTTGGGAGGTGGTCC
1375 SEQ ID NO: 116 -29.2 -40.6 91.7 -9.4 -1.9 -11.5
TCTCTGGAACACTGGTCGAC
525 SEQ ID NO: 117 -29.1 -39.2 89 -9.4 -0.4 -7.5
TGCAGAGTTCGATGAAATCT
975 SEQ ID NO: 118 -29.1 -33.4 80.9 -3.6 -0.2 -8.8
ATCCATCAGCATTTCTTTGA
4174 SEQ ID NO: 119 -29.1 -31.7 78.6 -2.6 0 -2.7
AGTTCCCGCCGCAGCCGAGA
64 SEQ ID NO: 120 -29 -45.2 97.5 -15.7 0 -7.7
GTCATCTCCAGATCCTTGGC
1232 SEQ ID NO: 121 -29 -39.1 87.9 -8.6 -1.4 -7
GTCTCATGCTGGGGCTTGAA
1311 SEQ ID NO: 122 -29 -39.6 90 -5.4 -5 -17.7
CAGAGCACACCAGGCAGAGT
1389 SEQ ID NO: 123 -29 -41.7 92.9 -11.8 -0.8 -5.4
TCCACCCCAGAGCAAATGCC
1944 SEQ ID NO: 124 -29 -41.7 93.1 -11.9 -0.6 -6.1
GGTCGTACATGCAGGGTAGA
2037 SEQ ID NO: 125 -29 -39.4 90.5 -9.6 0 -9.3
TTCTTGCCATATAGCCATTG
3420 SEQ ID NO: 126 -29 -32.9 82 -3.9 0 -5.6
GGCCACCTTGAATAGAAATC
3905 SEQ ID NO: 127 -29 -34.1 83.1 -4.3 0 -9.3
CACTGGGTCAGAGCCTCAGC
4313 SEQ ID NO: 128 -29 -43.1 94 -9.1 -5 -13.2 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecul Site oligo binding formation Duplex structure oligo oligo
CGAGGAAAGGCTGATTTGGC
455 SEQ ID NO: 129 -28.9 -37.4 87.6 -7.6 -0.6 -8.5
AACTCTTGGGGTTCTCTGGA
537 SEQ ID NO: 130 -28.8 -38.4 89.1 -6.6 -3 -11.9
GAAGACATTTTCGATAGCGG
1560 SEQ ID NO: 131 -28.8 -32.6 80.7 -3.2 -0.3 -7.4
GTTTCTTGTGAGACTCCTGT
1645 SEQ ID NO: 132 -28.8 -35 84.2 -2.1 -4.1 -10.4
AGGTGCCATCCTTCTTTGAC
2751 SEQ ID NO: 133 -28.8 -37.2 87.2 -6.8 -1.1 -10.8
TCATCCACAGTTTACCCAGC
3331 SEQ ID NO: 134 -28.8 -37.4 88 -8.6 0 -1.2
GCAACCATCATCCACAGTTT
3338 SEQ ID NO: 135 -28.8 -34.8 83.9 -6 0 -2.7
AGAGATTTAGTTTTTGGGTA
3576 SEQ ID NO: 136 -28.8 -29.2 77.5 0.4 0 -6.1
CTTGGGGCAGTGTTACATTA
939 SEQ ID NO: 137 -28.7 -34.8 85.9 -3.8 -1.4 -12.6
TGCCCAGTTTCTCTTGCTTA
1008 SEQ ID NO: 138 -28.7 -35.9 86.9 -7.2 0 -2.8
TCATGCTGGGGCTTGAATAG
1308 SEQ ID NO: 139 -28.7 -37.4 87.6 -5.4 -3.3 -12.8
TGAAGACATTTTCGATAGCG
1561 SEQ ID NO: 140 -28.7 -31.4 78.9 -2.1 -0.3 -7.4
TGGTCGTACATGCAGGGTAG
2038 SEQ ID NO: 141 -28.7 -39.1 90.6 -9.6 0 -9.4
GCCCTCTATAAACCACATGT
2605 SEQ ID NO: 142 -28.7 -35.3 85.6 -6.6 0 -5.2
TTTGCAACCATCATCCACAG
3341 SEQ ID NO: 143 -28.7 -34.7 83.8 -6 0 -6.2
GGAAAGGCTGATTTGGCCCT
452 SEQ ID NO: 144 -28.6 -39.2 90.5 -5.8 -4.7 -16.9
CTCATGCTGGGGCTTGAATA
1309 SEQ ID NO: 145 -28.6 -37.4 87.6 -5.4 -3.4 -13.6
GCTGGGGCTTGAATAGCCAT
1304 SEQ ID NO: 146 -28.5 -39.6 90.8 -5.4 -5.7 -14.7
ACATTTTCGATAGCGGCATG
1556 SEQ ID NO: 147 -28.5 -33.5 82.2 -4.5 0 -7.5
CAGTTTACCCAGCAGGTCAC
3324 SEQ ID NO: 148 -28.5 -38.2 89.7 -8.8 -0.7 -6.4
ATCCACAGTTTACCCAGCAG
3329 SEQ ID NO: 149 -28.5 -37.1 88.1 -8.6 0 -2.7
CATCCACAGTTTACCCAGCA
3330 SEQ ID NO: 150 -28.5 -37.1 88 -8.6 0 -2.7
TAGTCTTTTGCAACCATCAT
3347 SEQ ID NO: 151 -28.5 -31.5 79.8 -3 0 -6.2
TAGAGATTTAGTTTTTGGGT
3577 SEQ ID NO: 152 -28.5 -29.2 77.5 0.4 -0.3 -8.3
ACTGTTTGGTTTCTGAGACC
4241 SEQ ID NO: 153 -28.5 -34.7 84.4 -2.7 -1.8 -15.1
AGAGCCTCAGCAACCTTCAC
4304 SEQ ID NO: 154 -28.5 -39.5 90.3 -10.1 -0.7 -7.9 .
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TAATCTCACTGGGTCAGAGC
4319 SEQ ID NO: 155 -28.5 -37.9 87.6 -7.5 -0.1 -11.9
AGGAAAGGCTGATTTGGCCC
453 SEQ ID NO: 156 -28.4 -39.2 90.5 -5.8 -4.9 -17.1
CACCGTTGGTGCCAGTCTGG
648 SEQ ID NO: 157 -28.4 -41.7 93.5 -6.3 -4.8 -22.2
GGCAGAGTTTGGGAGGTGGT
1377 SEQ ID NO: 158 -28.4 -41.6 93.5 -13.2 0 -7.2
CTAGGTGCTCTATACCAGTT
3300 SEQ ID NO: 159 -28.4 -35.5 86.4 -4.2 -2.9 -8.4
AGTCTTTTGCAACCATCATC
3346 SEQ ID NO: 160 -28.4 -32.6 80.6 -4.2 0 -6.2
TTTCTTGCCATATAGCCATT
3421 SEQ ID NO: 161 -28.4 -31.7 80.5 -3.3 0 -5.6
CTGGGTCAGAGCCTCAGCAA
4311 SEQ ID NO: 162 -28.4 -41.8 93 -8.9 -4.5 -12
AATGAGAAGGGTGGTCAGAA
4365 SEQ ID NO: 163 -28.4 -36.2 85.7 -7.8 0 -5.5
GAGGAAAGGCTGATTTGGCC
454 SEQ ID NO: 164 -28.3 -38.3 88.7 -5.8 -4.1 -15.5
GAATCGTCTTCTCCCGCCAG
814 SEQ ID NO: 165 -28.3 -39.5 89.1 -11.2 0 -2.4
CTGTTGAGAAAGGGATGCTG
1145 SEQ ID NO: 166 -28.3 -35.9 85.4 -7 -0.3 -4.2
CTGAAGACATTTTCGATAGC
1562 SEQ ID NO: 167 -28.3 -31.1 78.5 -2.1 -0.4 -8.2
ATCTCCACCCCAGAGCAAAT
1947 SEQ ID NO: 168 -28.3 -38.5 88.6 -9.7 -0.1 -3
TTGACAAGAATACTGGAGAT
3200 SEQ ID NO: 169 -28.3 -32 79.6 -3.7 0 -2.8
AACCATCATCCACAGTTTAC
3336 SEQ ID NO: 170 -28.3 -32.8 81.5 -4.5 0 -0.6
TTAGTCTTTTGCAACCATCA
3348 SEQ ID NO: 171 -28.3 -31.3 80 -3 0 -6.2
CCGCTCGCCCGCCACCGTCC
86 SEQ ID NO: 172 -28.2 -48.9 101.2 -20 -0.5 -5.3
AAAGGCTGATTTGGCCCTGC
450 SEQ ID NO: 173 -28.2 -39 90.6 -5.8 -4.9 -17.1
TGGAACACTGGTCGACCTAT
521 SEQ ID NO: 174 -28.2 -38 88.4 -5.6 -0.8 -16.5
GGGTTCTCTGGAACACTGGT
529 SEQ ID NO: 175 -28.2 -39.5 90.4 -7.8 -3.5 -12.5
GTTTTCACTTGGGGCAGTGT
946 SEQ ID NO: 176 -28.2 -36.6 87.8 -6.7 -1.7 -6.4
AGTTGTCATCTCCAGATCCT
1236 SEQ ID NO: 177 -28.2 -36.7 85.2 -8.5 0 -4.8
AGGAGAGCTTACATCTGGTC
1328 SEQ ID NO: 178 -28.2 -37.9 87.6 -9.1 -0.3 -7.9
ATCAGAGCACACCAGGCAGA
1391 SEQ ID NO: 179 -28.2 -40.9 91.3 -11.8 -0.8 -5.4
TCATCAGAGCACACCAGGCA
1393 SEQ ID NO: 180 -28.2 -40.9 91.3 -11.8 -0.8 -5.4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TCCTGTAGTGGCCTGCTGAA
1631 SEQ ID NO: 181 -28.2 -40.8 92.7 -9.8 -2.4 -13.4
GGTGCCATCCTTCTTTGACT
2750 SEQ ID NO: 182 -28.2 -37.2 87.2 -7.5 -0.6 -10.8
AGGGCGTTAGGTACAGACAG
3393 SEQ ID NO: 183 -28.2 -38.9 91 -10.7 0.2 -7
CATCAGCATTTCTTTGACCC
4171 SEQ ID NO: 184 -28.2 -33.7 82.2 -5.5 0 -2.7
GGAGCTGGATGGAGGAGAGC
1340 SEQ ID NO: 185 -28.1 -43.7 93.5 -15.6 0 -5.3
TTTGACAAGAATACTGGAGA
3201 SEQ ID NO: 186 -28.1 -31.8 79.9 -3.7 0 -2.9
CGCAGCCGAGATAAACAACT
55 SEQ ID NO: 187 -28 -35.6 85.8 -7.6 0 -3.8
CGCCGCAGCCGAGATAAACA
58 SEQ ID NO: 188 -28 -39.5 90.8 -11 0 -7.5
GTTCCCGCCGCAGCCGAGAT
63 SEQ ID NO: 189 -28 -44.2 95.5 -15.7 0 -7.9
CTGGTCTCATGCTGGGGCTT
1314 SEQ ID NO: 190 -28 -41.7 92.9 -7.7 -5.7 -19.8
GAGTTTCCTTCCCTCTTGAC
2236 SEQ ID NO: 191 -28 -36.3 85.8 -7.7 -0.3 -4.2
TCTTGCCATATAGCCATTGC
3419 SEQ ID NO: 192 -28 -35.4 85.2 -7.4 0 -5.6
TATCCATCAGCATTTCTTTG
4175 SEQ ID NO: 193 -28 -30.6 77.7 -2.6 0 -2.7
AGAATGGGAGGCAGAGGATA
4349 SEQ ID NO: 194 -28 -39.1 89.1 -11.1 0 -2.4
GGAGCCGCTCGCCCGCCACC
90 SEQ ID NO: 195 -27.9 -49.8 102 -18.4 -3.2 -14.6
TCATCTCCAGATCCTTGGCA
1231 SEQ ID NO: 196 -27.9 -39 88.1 -9.6 -1.4 -7
GTTGTCATCTCCAGATCCTT
1235 SEQ ID NO: 197 -27.9 -35.5 83.6 -7.6 0 -4.8
CATCAGAGCACACCAGGCAG
1392 SEQ ID NO: 198 -27.9 -40.6 91 -11.8 -0.8 -5.4
TTCATCAGAGCACACCAGGC
1394 SEQ ID NO: 199 -27.9 -39.7 89.8 -11.8 0 -3.4
GGAGTTTCCTTCCCTCTTGA
2237 SEQ ID NO: 200 -27.9 -37.4 87.4 -7.7 -1.8 -9.2
TAGGTGCCATCCTTCTTTGA
2752 SEQ ID NO:201 -27.9 -36.3 86.5 -6.8 -1.3 -10.8
GGGCACTGGTGGTTTAGGTG
2766 SEQ ID NO: 202 -27.9 -40.3 92.8 -10.4 -1.7 -11.6
AAGAATACTGGAGATTTGAG
3195 SEQ ID NO: 203 -27.9 -30.7 78 -2.8 0 -4
CAAGAATACTGGAGATTTGA
3196 SEQ ID NO: 204 -27.9 -30.7 77.9 -2.8 0 -3.4
CTTTCTTGCCATATAGCCAT
3422 SEQ ID NO:205 -27.9 -32.9 82.1 -5 0 -5.6
GCCGCTCGCCCGCCACCGTC
87 SEQ ID NO: 206 -27.8 -49 101.1 -20.5 -0.5 -5.3 -
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
TCTTTGGAGTCCATCAGTGA 127 SEQ ID NO: 207 -27.8 -36.5 85.4 -6.3 0 -13
AGAGCACACCAGGCAGAGTT 1388 SEQ ID NO:208 -27.8 -40.5 91.9 -11.8 -0.8 -5.4
TTGGTCGTACATGCAGGGTA 2039 SEQ ID NO: 209 -27.8 -37.9 89.4 -9.6 0 -7.5
GACTGTGGAGATTACGTCCA
2734 SEQ ID NO:210 -27.8 -37.1 86.6 -7.7 -1.6 -7.9 TGACTGTGGAGATTACGTCC
2735 SEQ ID NO: 211 -27.8 -37.1 86.6 -7.7 -1.6 -7.9 ATTGGTGACAGATGGGAATG
3629 SEQ ID NO: 212 -27.8 -35.1 83.5 -4.2 -3.1 -6.2
ATGGGAGGCAGAGGATAACT 4346 SEQ ID N0:213 -27.8 -38.9 89.3 -11.1 0 -2.4
TGGTCAGAATGGGAGGCAGA 4354 SEQ ID NO: 214 -27.8 -41 91.5 -12.3 -0.8 -4.8
CAGGGGTGCAGAGTTCGATG 981 SEQ ID NO:215 -27.7 -39.9 90.1 -11.2 -0.9 -4.8
GCGTAGTCATGATCCTCCAA 1800 SEQ ID NO: 216 -27.7 -37.2 86.3 -8.8 0 -9.1
AGTCTTGGCCCTCTATAAAC 2612 SEQ ID NO: 217 -27.7 -35.3 85.9 -7.1 0 -7.5
CCCGCCGCAGCCGAGATAAA 60 SEQ ID NO: 218 -27.6 -41.8 93.8 -13.7 0 -7.5
GTCTTCGCTGCTTGGAGTCT 294 SEQ ID NO -.219 -27.6 -38.9 89.1 -8.5 -2.8 -10.4
GCAGAGTTCGATGAAATCTT 974 SEQ ID NO: 220 -27.6 -32.2 79.4 -3.6 -0.3 -9.7
ATGACGACTCAACTGCTTCT 2639 SEQ ID NO: 221 -27.6 -35.5 84.4 -7.9 0 -2.6
GGTTTAGGTGCCATCCTTCT 2756 SEQ ID NO: 222 -27.6 -37.3 88.2 -6.8 -2.9 -11.8
TGGGAGGCAGAGGATAACTT 4345 SEQ ID NO: 223 -27.6 -38.7 89.6 -11.1 0 -2.4
TGATTTGGCCCTGCTGTGGG 444 SEQ ID NO: 224 -27.5 -41.4 93.2 -10.5 -3.4 -11.5
GCCACCGTTGGTGCCAGTCT 650 SEQ ID NO: 225 -27.5 -43 95.2 -12.1 -3.2 -14.2
GCCCAGTTTCTCTTGCTTAA 1007 SEQ ID NO: 226 -27.5 -34.7 85.3 -7.2 0 -2.7
GCTGTTGAGAAAGGGATGCT 1146 SEQ ID NO: 227 -27.5 -37.2 87.1 -9 -0.5 -3.8
AGTTTCCTTCCCTCTTGACA 2235 SEQ ID NO: 228 -27.5 -36 86.1 -8.5 0 -2.9
TTGGGCACTGGTGGTTTAGG 2768 SEQ ID NO: 229 -27.5 -39 91.7 -9.4 -0.9 -12.3
TTTTGCAACCATCATCCACA 3342 SEQ ID NO: 230 -27.5 -33.5 82.3 -6 0 -6.2
TGGCAGACATTTTATTACCA 1068 SEQ ID NO:23l -27.4 -31.6 80.7 -4.2 0 -3.1
GTTGAGAAAGGGATGCTGTA 1143 SEQ ID NO:232 -27.4 -35.2 84.8 -7.8 0 -4.2 „
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
GATCTCCACCCCAGAGCAAA 1948 SEQ ID NO:233 -27.4 -39.8 90 -11.9 -0.1 -4.2
TGGGTCAGAGCCTCAGCAAC 4310 SEQ ID NO:234 -27.4 -41.9 93 -12.2 -2.3 -10.6
TCCCGCCGCAGCCGAGATAA 61 SEQ ID NO: 235 -27.3 -43.3 95.2 -15.5 0 -7.5
TCTGGTCTCATGCTGGGGCT 1315 SEQ ID NO: 236 -27.3 -43.2 94.3 -10.1 -5.6 -18.9
GGCCCTCTATAAACCACATG 2606 SEQ ID NO: 237 -27.3 -36.4 87 -8.6 0 -7.5
AGAATACTGGAGATTTGAGT 3194 SEQ ID NO: 238 -27.3 -32 79.7 -4.7 0 -6.6
ATTAGTCTTTTGCAACCATC 3349 SEQ ID NO: 239 -27.3 -30.3 78.2 -3 0 -6.2
TCCCTTCCCAGATTAGTGAA 4029 SEQ ID NO: 240 -27.3 -36.5 86.5 -9.2 0 -3
GCCTCAGCAACCTTCACTGC 4301 SEQ ID NO: 241 -27.3 -40.5 91.6 -12.4 -0.6 -3.8
GCAGCCGAGATAAACAACTT 54 SEQ ID NO: 242 -27.2 -34.1 83.9 -6.9 0 -3.2
GAAAGGCTGATTTGGCCCTG 451 SEQ ID NO: 243 -27.2 -38 88.8 -5.8 -4.9 -17.1
TTCTGTTTTCACTTGGGGCA 950 SEQ ID NO: 244 -27.2 -35.5 86.3 -7.8 -0.1 -6.4
TTATATTTGCTCCAGGAAAG 1047 SEQ ID NO: 245 -27.2 -30.5 78.9 -2.8 0 -7.6
TGTCATCTCCAGATCCTTGG 1233 SEQ ID NO: 246 -27.2 -37.8 86.5 -9.5 -1 -6.2
GCTTACATCTGGTCTCATGC 1322 SEQ ID NO: 247 -27.2 -36.7 85.8 -9.5 0 -5.5
AAACCAGACAGTAATAGCTA 2967 SEQ ID NO: 248 -27.2 -31.9 81.7 -4.7 0 -5.7
GTGAAGACGCAGAAACCTTC 249 SEQ ID NO: 249 -27.1 -35.3 84.7 -7 -1.1 -4.8
TGTTTTCACTTGGGGCAGTG 947 SEQ ID NO: 250 -27.1 -36.5 87.7 -8.1 -1.2 -7.1
GAGGAGCTGGATGGAGGAGA 1342 SEQ ID NO: 251 -27.1 -42.7 92.3 -15.6 0 -5.3
TGGTTTAGGTGCCATCCTTC 2757 SEQ ID NO: 252 -27.1 -37.3 88.1 -6.8 -3.4 -11.8
TCTCCACTGAAGCAGATATA 3250 SEQ ID NO: 253 -27.1 -34.8 83.5 -7.2 -0.1 -7.2
TAGGTACAGACAGGGCCTCT 3386 SEQ ID NO: 254 -27.1 -41.2 93.5 -12.2 -1.9 -10.7
TAGGGCGTTAGGTACAGACA 3394 SEQ ID NO: 255 -27.1 -38.1 90.5 -10.5 -0.1 -7.3
CTGTTGGAATGAGAAGGGTG 4372 SEQ ID NO: 256 -27.1 -35.9 85.5 -8.8 0 -1.1
TGGAAATAAACTCTTGTTGT 4659 SEQ ID NO: 257 -27.1 -28.9 76.6 -1.8 0 -2.8
GCCGCAGCCGAGATAAACAA 57 SEQ ID NO: 258 -27 -38 89 -11 0 -6.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GGAACACTGGTCGACCTATT
520 SEQ ID NO: 259 -27 -36.8 86.9 -5.6 -0.8 -16.5
TTTCACATTGCCACCGTTGG
659 SEQ ID NO: 260 -27 -35.8 86.4 -8.3 -0.2 -7.3
GCAGAGTTTGGGAGGTGGTC
1376 SEQ ID NO: 261 -27 -40.7 91.6 -13.2 0 -7.7
ACATTGGTCGTACATGCAGG
2042 SEQ ID NO: 262 -27 -36.5 86.6 -9.5 0 -5.1
TGGAGTTTCCTTCCCTCTTG
2238 SEQ ID NO: 263 -27 -37.1 87.5 -8.3 -1.8 -9.2
CTGGAGTTTCCTTCCCTCTT
2239 SEQ ID NO: 264 -27 -37.1 87.5 -8.3 -1.8 -9.2
GTCTTGGCCCTCTATAAACC
2611 SEQ ID NO: 265 -27 -36.5 87.3 -9 0 -7.5
GGTACAGACAGGGCCTCTTG
3384 SEQ ID NO: 266 -27 -40.8 92.4 -12.2 -1.2 -10.7
TTGGTGACAGATGGGAATGT
3628 SEQ ID NO: 267 -27 -36.2 85.4 -6.4 -2.8 -5.6
GGGAGGCAGAGGATAACTTC
4344 SEQ ID NO: 268 -27 -39 89.2 -11.1 -0.8 -5.6
AATCGTCTTCTCCCGCCAGA
813 SEQ ID NO:269 -26.9 -39.5 89.5 -12.6 0 -2.9
CCTGTAGTGGCCTGCTGAAT
1630 SEQ ID NO:270 -26.9 -39.5 90.9 -9.8 -2.2 -13.6
ACCAGACAGTAATAGCTATA
2965 SEQ ID NO: 271 -26.9 -32.5 82.2 -4.7 0 -9.8
AACCAGACAGTAATAGCTAT
2966 SEQ ID NO: 272 -26.9 -32.1 81.5 -4.7 0 -7.9
TTAGAGATTTAGTTTTTGGG
3578 SEQ ID NO: 273 -26.9 -27.9 75.6 0.4 -0.8 -9.3
TCACTGGGTCAGAGCCTCAG
4314 SEQ ID NO: 274 -26.9 -42.1 92.7 -10.6 -4.6 -12.2
CTCACTGTTGGAATGAGAAG
4376 SEQ ID NO: 275 -26.9 -33.8 82.2 -5.4 -1.4 -9
ACATTGCCACCGTTGGTGCC
655 SEQ ID NO: 276 -26.8 -40.5 92.4 -10.7 -2.7 -13.6
AGGAGCTGGATGGAGGAGAG
1341 SEQ ID NO: 277 -26.8 -42.4 92.4 -15.6 0 -5.3
TGAGGAGCTGGATGGAGGAG
1343 SEQ ID NO: 278 -26.8 -42.4 92.4 -15.6 0 -4.9
AGGCAGAGTTTGGGAGGTGG
1378 SEQ ID NO:279 -26.8 -41.5 93.4 -14.7 0 -7.2
TATGATCTCCACCCCAGAGC
1951 SEQ ID NO:280 -26.8 -40.4 90.3 -13.1 -0.1 -4.2
AAAACCAGACAGTAATAGCT
2968 SEQ ID NO:281 -26.8 -31.5 81 -4.7 0 -2.9
TATAATTCTCCACTGAAGCA
3256 SEQ ID NO: 282 -26.8 -32.1 80.4 -4.8 -0.2 -2.7
AGGTACAGACAGGGCCTCTT
3385 SEQ ID NO: 283 -26.8 -40.8 92.9 -12.2 -1.7 -10.7
GGGCGTTAGGTACAGACAGG
3392 SEQ ID NO: 284 -26.8 -40.1 92.3 -12.8 -0.1 -7.3 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
TTATCCATCAGCATTTCTTT 4176 SEQ ID NO: 285 -26.8 -29.4 76.1 -2.6 0 -2.7
AACTGTTTGGTTTCTGAGAC 4242 SEQ ID NO: 286 -26.8 -32.3 81.2 -4.9 0 -8.5
GAATGGGAGGCAGAGGATAA 4348 SEQ ID NO:287 -26.8 -37.9 87.6 -11.1 0 -2.4
GATTTGGCCCTGCTGTGGGA 443 SEQ ID NO:288 -26.7 -41.7 93.1 -10.5 -4.5 -13.8
TACTGGAGATTTGAGTCAAT
3190 SEQ ID NO:289 -26.7 -32 79.6 -3.7 -0.7 -11 ATACTGGAGATTTGAGTCAA
3191 SEQ ID NO: 290 -26.7 -32 79.6 -3.7 -0.7 -11 TTCTCCACTGAAGCAGATAT
3251 SEQ ID NO: 291 -26.7 -34.4 82.8 -7.2 -0.2 -7.2
GGACTAGGTGCTCTATACCA 3303 SEQ ID NO: 292 -26.7 -38.2 89.4 -8.6 -2.9 -11.8
CAACCATCATCCACAGTTTA 3337 SEQ ID NO: 293 -26.7 -32.7 81.4 -6 0 -0.6
AATTGGTGACAGATGGGAAT 3630 SEQ ID NO: 294 -26.7 -33.9 82.1 -4.1 -3.1 -6.2
GCCACCTTGAATAGAAATCA 3904 SEQ ID NO: 295 -26.7 -32.9 81.5 -6.2 0 -1.3
CTGTTTGGTTTCTGAGACCA 4240 SEQ ID NO: 296 -26.7 -34.6 84.3 -2.7 -3.4 -18.5
TTCCCGCCGCAGCCGAGATA 62 SEQ ID NO: 297 -26.6 -43.3 95.2 -16.2 0 -7.8
CAGTTCCCGCCGCAGCCGAG 65 SEQ ID NO: 298 -26.6 -44.9 96.9 -17.8 0 -7.5
TTCACATTGCCACCGTTGGT 658 SEQ ID NO:299 -26.6 -37.1 88 -8.3 -2.2 -11.3
CAGAGTTCGATGAAATCTTC 973 SEQ ID NO: 300 -26.6 -31.2 77.6 -3.6 -0.3 -9.7
GGAAAGCTTGCCTGACAGTA 1033 SEQ ID NO: 301 -26.6 -37.2 88.3 -8.6 -0.2 -12.1
AGAGTTTGGGAGGTGGTCCT 1374 SEQ ID NO: 302 -26.6 -40.6 92.2 -10.9 -3.1 -7
GAGCACACCAGGCAGAGTTT 1387 SEQ ID NO:303 -26.6 -39.3 90.4 -11.8 -0.8 -5.4
CTTCATCAGAGCACACCAGG 1395 SEQ ID NO: 304 -26.6 -38.4 88.2 -11.8 0 -2.7
CCACCCCAGAGCAAATGCCA 1943 SEQ ID NO:305 -26.6 -41.4 93.2 -14 -0.6 -6.1
TCTTGACAATGGCTTTTCCT 2223 SEQ ID NO: 306 -26.6 -33.5 82.6 -6.9 0 -3.8
CCCTCTATAAACCACATGTA 2604 SEQ ID NO: 307 -26.6 -33.2 83.1 -6.6 0 -6.9
TTTTGACAAGAATACTGGAG 3202 SEQ ID NO: 308 -26.6 -30.3 78.2 -3.7 0 -5.5
CAGGTCACTGGACTAGGTGC 3312 SEQ ID NO: 309 -26.6 -40.9 92.2 -11.3 -3 -10.7
TGGTGACAGATGGGAATGTG 3627 SEQ ID NO: 310 -26.6 -37.4 86.7 -8.6 -2.2 -4.4 _
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular inolecul Site oligo binding formation Duplex structure oligo oligo
CAATTGGTGACAGATGGGAA
3631 SEQ ID NO: 311 -26.6 -34.9 83.8 -4.1 -4.2 -8.4
AATGGGAGGCAGAGGATAAC
4347 SEQ ID NO: 312 -26.6 -37.7 87.7 -11.1 0 -2.4
ACTCACTGTTGGAATGAGAA
4377 SEQ ID NO: 313 -26.6 -33.9 82.4 -5.4 -1.9 -9.5
AGGAGCCGCTCGCCCGCCAC
91 SEQ ID NO: 314 -26.5 -48.6 101 -18.5 -3.4 -14.6
TTTCTGTTTTCACTTGGGGC
951 SEQ ID NO: 315 -26.5 -34.3 84.7 -7.8 0 -6.4
TGATCTCCACCCCAGAGCAA
1949 SEQ ID NO: 316 -26.5 -41 91.5 -14 -0.1 -4.2
CATTGGTCGTACATGCAGGG
2041 SEQ ID NO: 317 -26.5 -37.6 88 -11.1 0 -5.1
CAAGTCTTGGCCCTCTATAA
2614 SEQ ID NO: 318 -26.5 -35.2 85.7 -7.9 0 -9.4
GTTTAGGTGCCATCCTTCTT
2755 SEQ ID NO: 319 -26.5 -34.9 85.1 -6.8 -1.3 -10.6
ATTCTCCACTGAAGCAGATA
3252 SEQ ID NO: 320 -26.5 -34.4 82.8 -7.2 -0.5 -7.2
GGGTAAAGTTTAGTGAGAGG
3561 SEQ ID NO: 321 -26.5 -33.9 84.4 -7.4 0 -3.8
CAGAATGGGAGGCAGAGGAT
4350 SEQ ID NO: 322 -26.5 -39.9 89.6 -13.4 0 -2.4
TTTGGAGTCCATCAGTGAAT
125 SEQ ID NO: 323 -26.4 -34 82.2 -5.2 0 -13
ATTCTTTGGAGTCCATCAGT
129 SEQ ID NO: 324 -26.4 -34 82.2 -5.2 -0.2 -13
ATGGCAGACATTTTATTACC
1069 SEQ ID NO: 325 -26.4 -30.6 78.9 -4.2 0 -3.6
TCTCCAGATCCTTGGCACCT
1228 SEQ ID NO: 326 -26.4 -41 91.5 -13.1 -1.4 -7
TTAGGTGCCATCCTTCTTTG
2753 SEQ ID NO: 327 -26.4 -34.8 85 -6.8 -1.3 -10.8
TACAGACAGGGCCTCTTGGT
3382 SEQ ID NO: 328 -26.4 -40.8 92.9 -12.2 -2.2 -11.2
GTACAGACAGGGCCTCTTGG
3383 SEQ ID NO: 329 -26.4 -40.8 92.4 -12.2 -2.2 -11.2
AGTTTTTGGGTAAAGTTTAG
3568 SEQ ID NO: 330 -26.4 -27.3 76 -0.8 0 -3.8
CCGCAGCCGAGATAAACAAC
56 SEQ ID NO: 331 -26.3 -36.8 87.3 -10.5 0 -3.8
CCTCTTAGGGTTTTATAGAA
217 SEQ ID NO: 332 -26.3 -30.7 80.1 -3.5 -0.7 -8.4
AGGTGCTTTGGTCTGTGGTA
684 SEQ ID NO: 333 -26.3 -38.2 90.1 -11.9 0 -7.4
CTGTGCCCAGTTTCTCTTGC
1011 SEQ ID NO: 334 -26.3 -38 88.8 -11.1 -0.2 -7.9
AGGAAAGCTTGCCTGACAGT
1034 SEQ ID NO: 335 -26.3 -38 89.2 -8.6 -1.3 -14.3
CGTAGTCATGATCCTCCAAG
1799 SEQ ID NO: 336 -26.3 -35.9 84.7 -8.8 0 -9.4 _
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecu Site oligo binding formation Duplex structure oligo oligo
AAGTCTTGGCCCTCTATAAA
2613 SEQ ID NO: 337 -26.3 -34 84.4 -7.2 0 -7.5
TGACGACTCAACTGCTTCTG
2638 SEQ ID NO: 338 -26.3 -36.5 86 -10.2 0 -2.5
GTGCCATCCTTCTTTGACTG
2749 SEQ ID NO: 339 -26.3 -36 85.5 -9.7 0 -2.5
ACTGGAGATTTGAGTCAATT
3189 SEQ ID NO: 340 -26.3 -31.6 78.9 -3.7 -0.7 -11
TGGCCACCTTGAATAGAAAT
3906 SEQ ID NO: 341 -26.3 -33.8 83.3 -6.2 0 -10.5
TGCTTTGGTCTGTGGTATAC
681 SEQ ID NO: 342 -26.2 -35.2 85.7 -8.4 -0.1 -8.2
TCTGTTTTCACTTGGGGCAG
949 SEQ ID NO: 343 -26.2 -36.7 87.6 -7.8 -2.7 -10.9
GTGCCCAGTTTCTCTTGCTT
1009 SEQ ID NO: 344 -26.2 -36.8 87.6 -10.6 0 -2.7
GAAAGCTTGCCTGACAGTAA
1032 SEQ ID NO: 345 -26.2 -34.8 85.2 -8.6 0 -6.9
ATTATATTTGCTCCAGGAAA
1048 SEQ ID NO: 346 -26.2 -29.5 77 -2.8 0 -7.6
GGCAGACATTTTATTACCAA
1067 SEQ ID NO: 347 -26.2 -30.4 79.1 -4.2 0 -2.4
AAGCTTCATCAGAGCACACC
1398 SEQ ID NO: 348 -26.2 -37.3 87 -7.8 -3.3 -9
AGTCATGATCCTCCAAGTTG
1796 SEQ ID NO: 349 -26.2 -35.2 83.6 -8.2 0 -9.4
TCAGAATGGGAGGCAGAGGA
4351 SEQ ID NO: 350 -26.2 -41.2 91.4 -15 0 -2.5
TGAGAAGGGTGGTCAGAATG
4363 SEQ ID NO:351 -26.2 -37.4 86.9 -11.2 0 -5.5
TGGGGGAGGGCTGTATGTGA
4518 SEQ ID NO: 352 -26.2 -43.3 95.4 -17.1 0 -4.9
TCTTAGGGTTTTATAGAAGT
215 SEQ ID NO: 353 -26.1 -29.6 78.3 -3.5 0 -6.5
TCCTCTTAGGGTTTTATAGA
218 SEQ ID NO: 354 -26.1 -32.2 81.7 -5 -1 -7.4
TGTTTCTGTCTCTCCCATAT
398 SEQ ID NO: 355 -26.1 -34.4 82.9 -8.3 0 0
CATTGCCACCGTTGGTGCCA
654 SEQ ID NO: 356 -26.1 -40.4 92.3 -10.7 -3.3 -15
AATGAATCGTCTTCTCCCGC
817 SEQ ID NO: 357 -26.1 -36.1 84.7 -10 0 -5.2
TGGGGCAGTGTTACATTACT
937 SEQ ID NO: 358 -26.1 -36.1 87.6 -8.1 -1.2 -11.7
GCTTGCCTGACAGTAAACTG
1028 SEQ ID NO: 359 -26.1 -35.8 86.6 -8.6 -1 -6.9
AGCTTGCCTGACAGTAAACT
1029 SEQ ID NO: 360 -26.1 -35.8 86.9 -8.6 -1 -6.7
CATCTCCAGATCCTTGGCAC
1230 SEQ ID NO: 361 -26.1 -38.8 87.9 -11.3 -1.3 -6.9
TGGGGCTTGAATAGCCATTA
1302 SEQ ID NO: 362 -26.1 -36.3 87.3 -4.5 -5.7 -14.1 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecul Site oligo binding formation Duplex structure oligo oligo
GCTGGAGTTTCCTTCCCTCT
2240 SEQ ID NO: 363 -26.1 -39.6 90.4 -11.7 -1.8 -9.2
AAAACCGCTGCCAGTTCTGG
2259 SEQ ID NO: 364 -26.1 -38 89.8 -10 -1.2 -11.8
TTGACTGTGGAGATTACGTC
2736 SEQ ID NO: 365 -26.1 -34.7 83.6 -7.7 -0.8 -6.3
TGGACTAGGTGCTCTATACC
3304 SEQ ID NO: 366 -26.1 -38.2 89.4 -10.8 -1.2 -8.4
GATTTAGTTTTTGGGTAAAG
3573 SEQ ID NO: 367 -26.1 -26.5 74 0 0 -3.6
GTTTCTGAGACCATCGCTGC
4233 SEQ ID NO: 368 -26.1 -37.9 87.2 -10.7 0 -10.1
GGTCAGAATGGGAGGCAGAG
4353 SEQ ID NO: 369 -26.1 -41 91.1 -14 -0.8 -3.9
CTCTTAGGGTTTTATAGAAG
216 SEQ ID NO: 370 -26 -29.5 78.2 -3.5 0 -6.7
GGGGTTCTCTGGAACACTGG
530 SEQ ID NO: 371 -26 -40.6 91.7 -10.7 -3.9 -13.5
CTGGGGCTTGAATAGCCATT
1303 SEQ ID NO: 372 -26 -37.1 87.9 -5.4 -5.7 -14.7
AGCACACCAGGCAGAGTTTG
1386 SEQ ID NO: 373 -26 -39 90.4 -12.1 -0.8 -5.9
ATGATCTCCACCCCAGAGCA
1950 SEQ ID NO: 374 -26 -41.2 91.1 -15.2 0 -4.2
TAAAACCAGACAGTAATAGC
2969 SEQ ID NO: 375 -26 -30.7 80 -4.7 0 -1.1
CACTGAAGCAGATATATACA
3246 SEQ ID NO:376 -26 -31.3 79.2 -4.8 -0.1 -7.2
ACCATCATCCACAGTTTACC
3335 SEQ ID NO: 377 -26 -35.2 84.7 -9.2 0 -1.2
CTTCCCCTAGAGCAAACTGT
4256 SEQ ID NO: 378 -26 -37.1 88.3 -11.1 0 -4.3
GAGCCTCAGCAACCTTCACT
4303 SEQ ID NO: 379 -26 -39.5 90.3 -12.6 -0.7 -7.9
GGGGGAGGGCTGTATGTGAA
4517 SEQ ID NO:380 -26 -42.1 93.9 -16.1 0 -4.9
AAGACGCAGAAACCTTCACA
246 SEQ ID NO: 381 -25.9 -35 84.9 -9.1 0 -2.5
TTGTTTCTGTCTCTCCCATA
399 SEQ ID NO: 382 -25.9 -34.2 83.2 -8.3 0 0
GTGCTTTGGTCTGTGGTATA
682 SEQ ID NO: 383 -25.9 -35.2 85.7 -7.5 -1.6 -11.2
GGTGCTTTGGTCTGTGGTAT
683 SEQ ID NO: 384 -25.9 -37.2 88.1 -9.7 -1.3 -10.6
TCATCTATCAACAGGTCTGA
775 SEQ ID NO: 385 -25.9 -34.7 82.7 -7.2 0 -11.1
TTGGGGCAGTGTTACATTAC
938 SEQ ID NO: 386 -25.9 -34.9 86 -6.7 -1.4 -12.6
TTACATCTGGTCTCATGCTG
1320 SEQ ID NO: 387 -25.9 -35.4 84.3 -9.5 0 -6
CTTACATCTGGTCTCATGCT
1321 SEQ ID NO:388 -25.9 -35.4 84.3 -9.5 0 -6 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
AGCTTACATCTGGTCTCATG
1323 SEQ ID NO: 389 -25.9 -35.4 84.3 -9.5 0 -5
TGGGAGGTGGTCCTGTTGTT
1368 SEQ ID NO: 390 -25.9 -40.4 92.3 -11.4 -3.1 -8.3
TTGGGAGGTGGTCCTGTTGT
1369 SEQ ID NO: 391 -25.9 -40.4 92.3 -11.4 -3.1 -8.5
AGTTTGGGAGGTGGTCCTGT
1372 SEQ ID NO: 392 -25.9 -40.4 92.3 -11.4 -3.1 -8.3
ACAGTTTACCCAGCAGGTCA
3325 SEQ ID NO: 393 -25.9 -38.2 90.1 -11.4 -0.7 -6.4
GGCGTTAGGTACAGACAGGG
3391 SEQ ID NO: 394 -25.9 -40.1 92.3 -13.7 -0.1 -7.3
GTTTTTGGGTAAAGTTTAGT
3567 SEQ ID NO: 395 -25.9 -27.4 76.2 -1.4 0 -3.8
ATTTAGTTTTTGGGTAAAGT
3572 SEQ ID NO: 396 -25.9 -26.3 73.9 0 0 -3.3
GGTGACAGATGGGAATGTGA
3626 SEQ ID NO: 397 -25.9 -37.7 86.6 -11.1 -0.5 -4.1
TTCCCTTCCCAGATTAGTGA
4030 SEQ ID NO: 398 -25.9 -36.5 86.5 -10.6 0 -3
GAACACTGGTCGACCTATTG
519 SEQ ID NO:399 -25.8 -35.6 85.1 -5.6 -0.8 -16.5
TCAAAGGTGCTTTGGTCTGT
688 SEQ ID NO: 400 -25.8 -35.6 86.2 -7.9 -1.9 -9.6
CCTGAAGACATTTTCGATAG
1563 SEQ ID NO: 401 -25.8 -31 78.4 -4.5 -0.4 -8.2
CAGACCGTCCTTAGGAACTG
2153 SEQ ID NO: 402 -25.8 -37.7 88.4 -10.2 -1.4 -11
ACAAGAATACTGGAGATTTG
3197 SEQ ID NO: 403 -25.8 -30.5 78 -4.7 0 -3.4
GTCTTTTGCAACCATCATCC
3345 SEQ ID NO: 404 -25.8 -33.8 82.3 -8 0 -6
AATTAGTCTTTTGCAACCAT
3350 SEQ ID NO:405 -25.8 -28.8 76.4 -3 0 -6.2
ACAGACAGGGCCTCTTGGTA
3381 SEQ ID NO:406 -25.8 -40.8 92.9 -12.2 -2.8 -11.2
TTGGAAATAAACTCTTGTTG
4660 SEQ ID NO: 407 -25.8 -27.6 74.7 -1.8 0 -2.8
TTGGAGTCCATCAGTGAATA
124 SEQ ID NO:408 -25.7 -34.4 82.9 -6.3 0 -13
CTGCGCATTGCTTACTGAGC
335 SEQ ID NO:409 -25.7 -37.5 88.1 -10.1 -0.6 -11.5
TGCCACCGTTGGTGCCAGTC
651 SEQ ID NO: 410 -25.7 -43 95.2 -13.1 -3.9 -16.2
AGTTCGATGAAATCTTCTTT
970 SEQ ID NO: 411 -25.7 -28.5 74.2 -2.8 0 -4.9
AGCTTCATCAGAGCACACCA
1397 SEQ ID NO:412 -25.7 -38.5 88.5 -9.5 -3.3 -9
CTCCTGTAGTGGCCTGCTGA
1632 SEQ ID NO: 413 -257 -42 93.7 -13.5 -2.2 -13.6
CAGACAGGGCCTCTTGGTAG
3380 SEQ ID NO: 414 -25.7 -40.7 92.3 -12.2 -2.8 -10.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular Site oligo binding formation Duplex structure oligo oligo
TTAGTTTTTGGGTAAAGTTT
3570 SEQ ID NO: 415 -25.7 -26.1 74 0 0 -2.2
TTTAGTTTTTGGGTAAAGTT
3571 SEQ ID NO:416 -25.7 -26.1 74 0 0 -2.5
TCTTCCCCTAGAGCAAACTG
4257 SEQ ID NO:417 -25.7 -37.3 88.2 -11.1 -0.2 -4.3
TGCGCATTGCTTACTGAGCC
334 SEQ ID NO:418 -25.6 -38.7 89.8 -11 -1.4 -12.3
GTCAAAGGTGCTTTGGTCTG
689 SEQ ID NO:419 -25.6 -35.6 85.9 -7.9 -2.1 -9.6
ACATCTGGTCTCATGCTGGG
1318 SEQ ID NO: 420 -25.6 -39.8 89.5 -11.7 -2.5 -8.1
AGGTTTCTTGTGAGACTCCT
1647 SEQ ID NO: 421 -25.6 -36.1 85.9 -5.4 -4.9 -17.6
GCCTTTGCCCATTTCACTGC
1852 SEQ ID NO: 422 -25.6 -37.9 88.8 -12.3 0 -2
TCCTGAAACCTGGTATTGCC
1869 SEQ ID NO: 423 -25.6 -37.3 88.1 -11 -0.4 -8.3
AAGGTTTCTAATTTCTGGGA
2708 SEQ ID NO: 424 -25.6 -31.4 80.2 -5.2 -0.3 -5.1
AAATTAGTCTTTTGCAACCA
3351 SEQ ID NO: 425 -25.6 -28.6 76.6 -3 0 -6.2
GGTAAAGTTTAGTGAGAGGA
3560 SEQ ID NO: 426 -25.6 -33 82.8 -7.4 0 -3.9
GAGACCATCGCTGCCTGTAT
4227 SEQ ID NO: 427 -25.6 -39.4 89.3 -13.8 0 -4.5
TTTGTTTCTGTCTCTCCCAT
400 SEQ ID NO:428 -25.5 -33.8 82.5 -8.3 0 0
CCAGGGGTGCAGAGTTCGAT
982 SEQ ID NO: 429 -25.5 -41.1 91.9 -14.6 -0.9 -6.4
AATTATATTTGCTCCAGGAA
1049 SEQ ID NO: 430 -25.5 -29.5 77 -3.5 0 -7.6
GGTTTCTTGTGAGACTCCTG
1646 SEQ ID NO: 431 -25.5 -36.1 85.6 -6 -4.6 -15.8
TGGTATTGCCTTTGCCCATT
1859 SEQ ID NO: 432 -25.5 -35.7 86.8 -7.4 -2.8 -10.7
TCATCCAGGTGTAAGTTCCT
1885 SEQ ID NO:433 -25.5 -36.4 86.5 -9.7 -1.1 -6.2
CTTGACAATGGCTTTTCCTA
2222 SEQ ID NO: 434 -25.5 -32.4 81.7 -6.9 0 -3.8
TGGCCCTCTATAAACCACAT
2607 SEQ ID NO: 435 -25.5 -36.4 87.3 -10.4 0 -7.5
TTTGACTGTGGAGATTACGT
2737 SEQ ID NO:436 -25.5 -33.2 82.1 -7.7 0 -5.1
CCATCATCCACAGTTTACCC
3334 SEQ ID NO: 437 -25.5 -36.3 86.1 -10.8 0 -1.2
TAGTTTTTGGGTAAAGTTTA
3569 SEQ ID NO:438 -25.5 -26.5 74.9 -0.9 0 -2.4
CCCAGATTAGTGAATACCAA
4023 SEQ ID NO: 439 -25.5 -33 82.3 -7.5 0 -2.6
ATCAGCATTTCTTTGACCCC
4170 SEQ ID NO: 440 -25.5 -34.9 84 -9.4 0 -2.7 .
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TTTGGCCCTGCTGTGGGAAT
441 SEQ ID NO:441 -25.4 -40.2 92.2 -10.5 -4.3 -13.8
GTTCGATGAAATCTTCTTTT
969 SEQ ID NO: 442 -25.4 -27.3 72.5 -1.9 0 -4.9
CAATTATATTTGCTCCAGGA
1050 SEQ ID NO: 443 -25.4 -30.7 78.7 -5.3 0 -5.4
ATCTCCAGATCCTTGGCACC
1229 SEQ ID NO: 444 -25.4 -40 89.6 -13.1 -1.4 -7
CATTTTCGATAGCGGCATGC
1555 SEQ ID NO: 445 -25.4 -34.7 83.7 -7.9 0 -10.8
TAGTCATGATCCTCCAAGTT
1797 SEQ ID NO: 446 -25.4 -34.4 82.9 -8.2 0 -9.4
GTCAGTTGATAAAACCGCTG
2269 SEQ ID NO: 447 -25.4 -32.9 82.2 -6.8 -0.4 -8.2
GACGACTCAACTGCTTCTGT
2637 SEQ ID NO: 448 -25.4 -36.6 86.1 -11.2 0 -2.5
TTTTTGACAAGAATACTGGA
3203 SEQ ID NO: 449 -25.4 -29.1 76.5 -3.7 0 -6.9
ATAGGGCGTTAGGTACAGAC
3395 SEQ ID NO:450 -25.4 -37.1 88.6 -11.7 0 -6.5
GCTTTCTTGCCATATAGCCA
3423 SEQ ID NO: 451 -25.4 -35.2 85.5 -8.9 -0.8 -6.9
TTTCTGAGACCATCGCTGCC
4232 SEQ ID NO: 452 -25.4 -39 88.9 -13.6 0 -6.2
TGTTGGAATGAGAAGGGTGG
4371 SEQ ID NO: 453 -25.4 -37.1 87.2 -11.7 0 -0.2
TCTGGCTGCTGCGCATTGCT
343 SEQ ID NO: 454 -25.3 -41.9 93.4 -13.1 -1.7 -15.1
ATTGCCACCGTTGGTGCCAG
653 SEQ ID NO: 455 -25.3 -40.4 92.3 -10.7 -4.1 -16.5
ATTTCACATTGCCACCGTTG
660 SEQ ID NO: 456 -25.3 -33.6 82.9 -8.3 0 -0.9
CAATTTCACATTGCCACCGT
662 SEQ ID NO: 457 -25.3 -33.6 82.9 -8.3 0 -1
ACAATTTCACATTGCCACCG
663 SEQ ID NO: 58 -25.3 -33.6 82.9 -8.3 0 -2.2
TTTGGGTTTAGTGTCCGGTA
881 SEQ ID NO:459 -25.3 -35.2 87 -6.2 -3.7 -7.4
AAGCTTGCCTGACAGTAAAC
1030 SEQ ID NO:460 -25.3 -34.6 85.3 -8.6 -0.4 -6.1
GAGGAGAGCTTACATCTGGT
1329 SEQ ID NO:461 -25.3 -37.9 87.6 -11.9 -0.3 -8.7
GAAGCTTCATCAGAGCACAC
1399 SEQ ID NO:462 -25.3 -36.4 85.2 -7.8 -3.3 -13.5
GTCATGATCCTCCAAGTTGA
1795 SEQ ID NO: 463 -25.3 -35.5 83.6 -9.7 0 -8
TGTTGAGCGTAGTCATGATC
1806 SEQ ID NO: 464 -25.3 -34.9 83.2 -8.8 0 -9.4
CCTTCCCTCTTGACAATGGC
2230 SEQ ID NO:465 -25.3 -38.3 88.6 -13 0 -3.8
CAGCTGTACAATAACTTGAA
3048 SEQ ID NO:466 -25.3 -30.3 79 -4.3 0 -8.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
TGGGTAAAGTTTAGTGAGAG 3562 SEQ ID NO: 467 -25.3 -32.7 82.8 -7.4 0 -3.8
AGACCATCGCTGCCTGTATG 4226 SEQ ID NO: 468 -25.3 -39.1 89.3 -13.8 0 -4.5
CCGTCCGCAGTTCCCGCCGC 72 SEQ ID NO: 469 -25.2 -46.5 98.8 -20.7 -0.3 -3.8
TTCTTTGGAGTCCATCAGTG 128 SEQ ID NO: 470 -25.2 -35 83.9 -7.5 0 -12.8
GATTCTTTGGAGTCCATCAG 130 SEQ ID NO: 471 -25.2 -34.2 82 -5.2 -3.8 -14.2
TGAAGACGCAGAAACCTTCA 248 SEQ ID NO: 472 -25.2 -35.2 84.8 -7.5 -2.5 -7.6
ATCTGGCTGCTGCGCATTGC 344 SEQ ID NO:473 -25.2 -40.9 91.6 -13.8 -1.6 -11.5
TTTTGTTTCTGTCTCTCCCA 401 SEQ ID NO: 474 -25.2 -33.6 82.8 -8.4 0 0
ATTTGGCCCTGCTGTGGGAA 442 SEQ ID NO: 475 -25.2 -40.2 92.2 -10.5 -4.5 -13.8
GAAGATACATCAGAGTGAGT 607 SEQ ID NO: 476 -25.2 -33.6 81.1 -8.4 0 -3.5
ACCGTTGGTGCCAGTCTGGC 647 SEQ ID NO: 477 -25.2 -43 95.2 -11.1 -4.3 -21.6
GTTTTCATCTATCAACAGGT 779 SEQ ID NO: 478 -25.2 -30.6 78.1 -5.4 0 -1.5
CCAAAAGGAATGAATCGTCT 825 SEQ ID NO: 479 -25.2 -31.8 79.5 -6 -0.3 -6.8
CCCAGTTTCTCTTGCTTAAT 1006 SEQ ID NO: 480 -25.2 -32.4 81.8 -7.2 0 -2.7
TGTGCCCAGTTTCTCTTGCT 1010 SEQ ID NO: 81 -25.2 -38 89.1 -12.8 0 -3.1
TGCTGTTGAGAAAGGGATGC 1147 SEQ ID NO: 482 -25.2 -37.2 87.1 -11.3 -0.5 -3.8
CATCTGGTCTCATGCTGGGG 1317 SEQ ID NO: 483 -25.2 -40.9 90.8 -12.5 -3.2 -9.9
GTATTGCCTTTGCCCATTTC 1857 SEQ ID NO: 484 -25.2 -33.6 83.3 -8.4 0 -3.3
GTGTTTACATTGGTCGTACA
2048 SEQ ID NO: 485 -25.2 -32 81.4 -6.8 0 -5.9 TGTGTTTACATTGGTCGTAC
2049 SEQ ID NO: 486 -25.2 -32 81.4 -6.8 0 -6.5 TCATACAGAGATACTCTTCA
2112 SEQ ID NO: 487 -25.2 -32.7 80.2 -6.6 -0.8 -7.9
TAAAACCGCTGCCAGTTCTG 2260 SEQ ID NO: 488 -25.2 -36 87.4 -10 0 -9.4
TTTAGGTGCCATCCTTCTTT 2754 SEQ ID NO: 489 -25.2 -33.6 83.6 -6.8 -1.3 -10.8
GTTATCCATCAGCATTTCTT 4177 SEQ ID NO: 490 -25.2 -30.7 77.9 -5.5 0 -2.7
TTCTGAGACCATCGCTGCCT 4231 SEQ ID NO: 491 -25.2 -40.2 90.4 -15 0 -6.2
GGTTTCTGAGACCATCGCTG 4234 SEQ ID NO: 492 -25.2 -37.8 87.2 -7.4 -3.5 -18.5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GGAAATAAACTCTTGTTGTA
4658 SEQ ID NO: 493 -25.2 -28.1 75.7 -2.9 0 -2.8
CTTTGGAGTCCATCAGTGAA
126 SEQ ID NO: 494 -25.1 -35 83.9 -7.5 0 -13
CTTGGAGTCTGATTGAGAAG
284 SEQ ID NO:495 -25.1 -34 82.1 -8.2 -0.1 -9.1
TTGGCCCTGCTGTGGGAATC
440 SEQ ID NO: 496 -25.1 -41.7 93.1 -12.1 -4.5 -13.5
TGAAGATACATCAGAGTGAG
608 SEQ ID NO: 497 -25.1 -33.5 80.9 -8.4 0 -3.5
ATGTCAAAGGTGCTTTGGTC
691 SEQ ID NO: 498 -25.1 -34.6 84.2 -7.4 -2.1 -9.6
AGTTTTCATCTATCAACAGG
780 SEQ ID NO: 499 -25.1 -30.5 78 -5.4 0 -1.5
TTGAGAAAGGGATGCTGTAT
1142 SEQ ID NO: 500 -25.1 -34.1 83.1 -9 0 -4.5
AGGTTTCTAATTTCTGGGAT
2707 SEQ ID NO: 501 -25.1 -31.6 80 -5.2 -1.2 -6.8
CTGGAGATTTGAGTCAATTT
3188 SEQ ID NO: 502 -25.1 -30.3 77.2 -3.7 -0.5 -10.8
TAGGTGCTCTATACCAGTTA
3299 SEQ ID NO: 503 -25.1 -34.7 85.8 -6.7 -2.9 -8.4
AGGGCCTCTTGGTAGTTATT
3375 SEQ ID NO: 504 -25.1 -36.2 87.7 -8.3 -2.8 -10.7
GAGGAGCCGCTCGCCCGCCA
92 SEQ ID NO: 505 -25 -48.8 100.8 -20.2 -3.4 -14.6
CTGATTGAGAAGCGACAGCC
276 SEQ ID NO: 506 -25 -37.8 87.2 -11.3 -1.4 -7
GAGTCTGATTGAGAAGCGAC
280 SEQ ID NO: 507 -25 -36 84.1 -9.2 -1.8 -6.1
AGTGTCCGGTAAAATGAGAG
872 SEQ ID NO: 508 -25 -34.3 83.7 -9.3 0 -6.2
TCATTCCAGCCTGAAGACAT
1572 SEQ ID NO: 509 -25 -36.4 85.3 -11.4 0 -6.8
ATATGATCTCCACCCCAGAG
1952 SEQ ID NO: 510 -25 -38.1 87.1 -13.1 0 -4.2
TACATTGGTCGTACATGCAG
2043 SEQ ID NO: 511 -25 -34.5 84.2 -9.5 0 -5.2
GTTGATAAAACCGCTGCCAG
2265 SEQ ID NO: 512 -25 -35 85.5 -10 0 -4.5
AGTTGATAAAACCGCTGCCA
2266 SEQ ID NO: 513 -25 -35 85.7 -10 0 -5.5
AGGAGATTTTCAACCACTTC
2311 SEQ ID NO: 514 -25 -32.6 80.9 -6.7 0 -9.8
GATGACGACTCAACTGCTTC
2640 SEQ JD NO: 515 -25 -35.8 84.1 -10.8 0 -2.6
AGCTGTACAATAACTTGAAT
3047 SEQ ID NO: 516 -25 -29.3 77.2 -4.3 0 -7.2
ACAGCTGTACAATAACTTGA
3049 SEQ ID NO: 517 -25 -31.6 80.8 -4.3 0 -12.7
TGGAGTCCATCAGTGAATAT
123 SEQ ID NO: 518 -24.9 -34.6 82.6 -7.7 0 -12.1 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
AGGGTGAAGACGCAGAAACC
252 SEQ ID NO: 519 -24.9 -38.6 89.5 -12.5 -1.1 -5.1
CTCTGGAACACTGGTCGACC
524 SEQ ID NO: 520 -24.9 -40.1 90.4 -12.2 -0.4 -14.1
TGTCAAAGGTGCTTTGGTCT
690 SEQ ID NO: 521 -24.9 -35.6 86.2 -8.6 -2.1 -9.5
TTGGGTTTAGTGTCCGGTAA
880 SEQ ID NO: 522 -24.9 -35.2 87 -6.8 -3.5 -7
CGTACATGCAGGGTAGAGTC
2034 SEQ ID NO: 523 -24.9 -38.2 88.8 -12.5 0 -9.4
GTTTCCTTCCCTCTTGACAA
2234 SEQ ID NO: 524 -24.9 -34.8 84.4 -9.9 0 -2.9
TTGGCCCTCTATAAACCACA
2608 SEQ ID NO: 525 -24.9 -36.2 87.6 -10.8 -0.1 -7.5
ACTGTGGAGATTACGTCCAC
2733 SEQ ID NO: 526 -24.9 -36.9 86.8 -7.7 -4.3 -11.6
ATTAGAGATTTAGTTTTTGG
3579 SEQ ID NO: 527 -24.9 -25.7 71.7 0.4 -0.5 -8.5
CCATATTAGAGATTTAGTTT
3583 SEQ ID NO: 528 -24.9 -26.3 72.3 -1.3 0 -6.3
CCCTTCCCAGATTAGTGAAT
4028 SEQ ID NO: 529 -24.9 -35.2 84.7 -10.3 0 -3
GGAGGCAGAGGATAACTTCC
4343 SEQ ID NO: 530 -24.9 -39 89.2 -11.1 -3 -9.4
CTCCAGGAAAGCTTGCCTGA
1038 SEQ ID NO: 531 -24.8 -39.3 90.4 -8.1 -4.6 -21
GACTCCTGTAGTGGCCTGCT
1634 SEQ ID NO: 532 -24.8 -42.1 93.8 -15.1 -2.2 -10.5
CCTCTATAAACCACATGTAG
2603 SEQ ID NO: 533 -24.8 -32 81.3 -6.6 -0.3 -6.9
ACTGAAGCAGATATATACAA
3245 SEQ ID NO: 534 -24.8 -30.1 77.6 -4.8 -0.1 -7.2
TAAATTAGTCTTTTGCAACC
3352 SEQ ID NO: 535 -24.8 -27.8 75.7 -3 0 -6.2
GAATTACTTTGTCTGATTAA
3542 SEQ ID NO: 536 -24.8 -26.1 71.7 -1.2 0 -3.4
GGAATTACTTTGTCTGATTA
3543 SEQ ID NO: 537 -24.8 -28.5 75.4 -3.7 0 -3.4
CTTGTGTTAAACTCTATGGC
3755 SEQ ID NO: 538 -24.8 -31.5 80.9 -6.7 0 -4.3
TTGGCCACCTTGAATAGAAA
3907 SEQ ID NO: 539 -24.8 -33.6 83.6 -7.2 0 -11.2
TTCCCCTAGAGCAAACTGTT
4255 SEQ ID NO: 540 -24.8 -35.9 87.1 -11.1 0 -4
CTGGGGGAGGGCTGTATGTG
4519 SEQ ID NO: 541 -24.8 -43 94.9 -18.2 0 -4.9
TGTTGTAGGATAGAAAGGAA
4645 SEQ ID NO: 542 -24.8 -31.9 80.8 -7.1 0 -6
TTGTTGTAGGATAGAAAGGA
4646 SEQ ID NO: 543 -24.8 -31.9 80.8 -7.1 0 -7.2
ATTTGGAAATAAACTCTTGT
4662 SEQ ID NO: 544 -24.8 -26.6 72.7 -1.8 0 -2.8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecu; Site oligo binding formation Duplex structure oligo oligo
TGGGTTTAGTGTCCGGTAAA
879 SEQ ID NO: 545 -24.7 -35.2 87 -7.3 -3.2 -6.4
CCAATTTTCGGAACCAACGG
1205 SEQ ID NO: 546 -24.7 -33.8 83.7 -8.5 -0.3 -6.4
TACATCTGGTCTCATGCTGG
1319 SEQ ID NO: 547 -24.7 -37.8 87.3 -11.9 -1.1 -6
GTTTGGGAGGTGGTCCTGTT
1371 SEQ ID NO: 548 -24.7 -39.2 90.8 -11.4 -3.1 -8.5
TCCCTCTTGACAATGGCTTT
2227 SEQ ID NO: 549 -24.7 -35.9 85.9 -11.2 0 -4.1
TTCCCTCTTGACAATGGCTT
2228 SEQ ID NO: 550 -24.7 -35.9 85.9 -11.2 0 -4.1
TAAGGTTTCTAATTTCTGGG
2709 SEQ ID NO: 551 -24.7 -30.3 79.3 -5.6 0 -4
TAACTATACAGGGGGGGGAT
2866 SEQ ID NO: 552 -24.7 -39.1 91.3 -14.4 0 -1.6
GGTCACTGGACTAGGTGCTC
3310 SEQ ID NO: 553 -24.7 -41.2 92.1 -13.4 -3.1 -12.3
TACTTTGTCTGATTAAAAGT
3538 SEQ ID NO: 554 -24.7 -26.9 73.6 -1.2 -0.8 -7.1
GACCATCGCTGCCTGTATGA
4225 SEQ ID NO: 555 -24.7 -39.4 89.2 -13.8 -0.7 -5.4
TGGTTTCTGAGACCATCGCT
4235 SEQ ID NO: 556 -24.7 -37.8 87.6 -7.4 -4 -19.5
GGGTGAAGACGCAGAAACCT
251 SEQ ID NO: 557 -24.6 -38.6 89.5 -11.7 -2.3 -9.3
CCAGTCTGGCCCTTCAAATG
637 SEQ ID NO: 558 -24.6 -38 88.6 -11.3 0 -12.4
GCTTTGGTCTGTGGTATACA
680 SEQ ID NO: 559 -24.6 -35.2 85.7 -10.6 0 -7.3
TTTGGGAGGTGGTCCTGTTG
1370 SEQ ID NO: 560 -24.6 -39.1 90.7 -11.4 -3.1 -8.5
GAGGTTTCTTGTGAGACTCC
1648 SEQ ID NO: 561 -24.6 -36.4 85.6 -6.9 -4.9 -13.8
GTGAGTTGTGGTAACGTTGC
1699 SEQ ID NO: 562 -24.6 -35.1 85.6 -8.8 -0.1 -11.5
TCAGTTGATAAAACCGCTGC
2268 SEQ ID NO: 563 -24.6 -34.1 83.8 -8.6 -0.8 -5.6
GGAGATTTTCAACCACTTCA
2310 SEQ ID NO: 564 -24.6 -32.6 80.9 -6.7 -1.1 -9.8
TGAAAAAAAGTATGAAGAGA
2806 SEQ ID NO: 565 -24.6 -26.9 72.1 -2.3 0 -1
TGAGTCAATTTTTGTGGTCT
3179 SEQ ID NO: 566 -24.6 -31.2 79.2 -6.6 0 -5.9
TTTGGAAATAAACTCTTGTT
4661 SEQ ID NO: 567 -24.6 -26.4 72.8 -1.8 0 -2
TTTCATCTATCAACAGGTCT
777 SEQ ID NO: 568 -24.5 -32 79.6 -7.5 0 -3.8
TTTTCATCTATCAACAGGTC
778 SEQ ID NO: 569 -24.5 -30.8 78 -6.3 0 -2
TTGTCATCTCCAGATCCTTG
1234 SEQ ID NO: 570 -24.5 -35.4 83.5 -10.9 0 -4.5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GAGCTGGATGGAGGAGAGCT
1339 SEQ ID NO: 571 -24.5 -42.5 92.4 -16.4 -1.6 -6.1
GTAGTCATGATCCTCCAAGT
1798 SEQ ID NO: 572 -24.5 -35.7 84.4 -10.4 0 -9.4
ATCCAGGTGTAAGTTCCTGA
1883 SEQ ID NO: 573 -24.5 -36.4 86.5 -7.1 -4.8 -12.2
ACCCCAGAGCAAATGCCATA
1941 SEQ ID NO: 574 -24.5 -38.4 89.5 -13.1 -0.6 -6.1
GTACATGCAGGGTAGAGTCA
2033 SEQ ID NO: 575 -24.5 -37.9 88.5 -12.5 -0.2 -9.4
GTCGTACATGCAGGGTAGAG
2036 SEQ ID NO: 576 -24.5 -38.2 88.8 -12.3 -0.2 -10.8
TCAACCACTTCATGCATAGA
2302 SEQ ID NO: 577 -24.5 -34.2 82.9 -9.7 0 -7
AAAGTATGAAGAGAAAGTTC
2800 SEQ ID NO: 578 -24.5 -28.2 74.8 -3.7 0 -2.6
AAACTGTTTGGTTTCTGAGA
4243 SEQ ID NO: 579 -24.5 -31 79.5 -5.6 -0.8 -7.3
GTAATCTCACTGGGTCAGAG
4320 SEQ ID NO: 580 -24.5 -36.7 86 -10.3 -0.1 -11.9
GACTCACTGTTGGAATGAGA
4378 SEQ ID NO: 581 -24.5 -35.4 83.7 -9 -1.9 -9.5
CAGCCGAGATAAACAACTTA
53 SEQ ID NO: 582 -24.4 -32 81.3 -7.6 0 -1.7
GGAGTCCATCAGTGAATATC
122 SEQ ID NO: 583 -24.4 -34.9 82.5 -8.9 0 -11.1
GGTGAAGACGCAGAAACCTT
250 SEQ ID NO: 584 -24.4 -36.2 86.5 -10.3 -1.4 -7.5
GGTCGACCTATTGAGGTTTG
512 SEQ ID NO: 585 -24.4 -35.5 85.3 -6.8 -3.8 -16.5
TCCAGGAAAGCTTGCCTGAC
1037 SEQ ID NO: 586 -24.4 -39.4 90.4 -8.6 -4.6 -21
CCTGAAACCTGGTATTGCCT
1868 SEQ ID NO: 587 -24.4 -37 88.2 -11 -1.1 -10.7
GATAAAACCGCTGCCAGTTC
2262 SEQ ID NO: 588 -24.4 -35.3 85.4 -10 0 -9.5
TTGATAAAACCGCTGCCAGT
2264 SEQ ID NO: 589 -24.4 -35 85.7 -10 0 -8.5
TCAGCTAACATCTCGGGGAA
2377 SEQ ID NO: 590 -24.4 -38.1 88.4 -12.3 0 -10.7
CCAATTGGTGACAGATGGGA
3632 SEQ ID NO: 591 -24.4 -37.3 86.8 -5.4 -7.5 -15
TCAGCATTTCTTTGACCCCT
4169 SEQ ID NO: 592 -24.4 -35.9 85.9 -11.5 0 -2.7
ACAGACTCACTGTTGGAATG
4381 SEQ ID NO: 593 -24.4 -34.9 83.9 -8.9 -1.5 -5.4
CCGAGGAAAGGCTGATTTGG
456 SEQ ID NO: 594 -24.3 -37.3 87.6 -10.9 -1 -12.4
TTACCAATTATATTTGCTCC
1054 SEQ ID NO: 595 -24.3 -28.5 76.2 -4.2 0 -4.6
CAATTTTCGGAACCAACGGG
1204 SEQ ID NO: 596 -24.3 -33.8 83.7 -8.5 -0.9 0 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CTCCAGATCCTTGGCACCTA
1227 SEQ ID NO: 597 -24.3 -39.9 90.7 -14.1 -1.4 -7
TTCGATAGCGGCATGCTGGG
1551 SEQ ID NO: 598 -24.3 -40.5 91.4 -11.3 -2.3 -17.9
CCTTTGCCCATTTCACTGCT
1851 SEQ ID NO: 599 -24.3 -36.6 87.4 -12.3 0 -1.8
GGTATTGCCTTTGCCCATTT
1858 SEQ ID NO: 600 -24.3 -34.5 85.2 -7.4 -2.8 -10.7
ATTGGTCGTACATGCAGGGT
2040 SEQ ID NO: 601 -24.3 -37.7 88.4 -13.4 0 -5.1
AGACCGTCCTTAGGAACTGA
2152 SEQ ID NO: 602 -24.3 -38 88.7 -12.1 -1.2 -11
TCAGACCGTCCTTAGGAACT
2154 SEQ ID NO: 603 -24.3 -38 88.7 -12.1 -0.7 -11
TGTCAGTTGATAAAACCGCT
2270 SEQ ID NO: 604 -24.3 -32.9 82.3 -7 -1.3 -10.4
CTCCACTGAAGCAGATATAT
3249 SEQ ID NO: 605 -24.3 -33.5 81.7 -9.2 0.4 -6.7
ATATAATTCTCCACTGAAGC
3257 SEQ ID NO: 606 -24.3 -31.1 78.7 -6.1 -0.5 -2.6
TAGGACTGTTAATTTGCACA
3281 SEQ ID NO: 607 -24.3 -31.5 80.9 -6.7 -0.1 -5.4
AGGTGCTCTATACCAGTTAG
3298 SEQ ID NO: 608 -24.3 -35.5 86.4 -8.3 -2.9 -8.4
CATCATCCACAGTTTACCCA
3333 SEQ ID NO: 609 -24.3 -35.1 84.6 -10.8 0 -1.2
TTACTTTGTCTGATTAAAAG
3539 SEQ ID NO: 610 -24.3 -25.6 71.8 -1.2 0 -4.3
GTTTCTGTCTCTCCCATATA
397 SEQ ID NO: 611 -24.2 -33.6 82.1 -9.4 0 -0.7
TCCCATCACTTTTGTTTCTG
410 SEQ ID NO: 612 -24.2 -32.3 80.9 -8.1 0 0
TGGCCCTGCTGTGGGAATCC
439 SEQ ID NO: 613 -24.2 -44.1 95.8 -15.4 -4.5 -13.8
TACAATTTCACATTGCCACC
664 SEQ ID NO: 614 -24.2 -32.5 81.6 -8.3 0 -2.2
ATCGTCTTCTCCCGCCAGAG
812 SEQ ID NO: 615 -24.2 -40.7 90.6 -16 -0.2 -5.7
TCCAAAAGGAATGAATCGTC
826 SEQ ID NO: 616 -24.2 -32.1 79.5 -6 -1.9 -9.2
TAGTGTCCGGTAAAATGAGA
873 SEQ ID NO: 617 -24.2 -33.5 83 -9.3 0 -6.3
TGAGAAAGGGATGCTGTATT
1141 SEQ ID NO: 618 -24.2 -34.1 83.1 -9.9 0 -7.1
TGAGTTGTGGTAACGTTGCA
1698 SEQ ID NO: 619 -24.2 -35 85.8 -7.6 -1.3 -14.6
TATATGATCTCCACCCCAGA
1953 SEQ ID NO: 620 -24.2 -37.3 86.5 -13.1 0 -4.2
ATAAAACCGCTGCCAGTTCT
2261 SEQ ID NO: 621 -24.2 -35 85.8 -10 0 -9.4
TGATAAAACCGCTGCCAGTT
2263 SEQ ID NO: 622 -24.2 -35 85.7 -10 0 -9.4 -
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
TAAACCACATGTAGTGCGTA 2597 SEQ ID NO: 623 -24.2 -33.3 83.9 -6.7 -2.4 -10.2
AGACAGTAATAGCTATAAAA 2962 SEQ ID NO: 624 -24.2 -27.6 75.2 -2.4 0 -9.8
GCTGTACAATAACTTGAATA 3046 SEQ ID NO: 625 -24.2 -28.5 76.2 -4.3 0 -7.2
TGCTCTATACCAGTTAGGAC 3295 SEQ ID NO: 626 -24.2 -35.8 86.4 -10.7 -0.8 -4.6
AGGTCACTGGACTAGGTGCT 3311 SEQ ID NO: 627 -24.2 -40.9 92.7 -13.4 -3.3 -12.7
ACCGTCCGCAGTTCCCGCCG 73 SEQ ID NO: 628 -24.1 -45.3 97.9 -20.7 -0.1 -3.1
GCTCGCCCGCCACCGTCCGC 84 SEQ ID NO: 629 -24.1 -49 101.1 -24.2 -0.4 -4.7
TGATTCTTTGGAGTCCATCA 131 SEQ ID NO: 630 -24.1 -34.2 82.1 -6.3 -3.8 -14.2
TCACAGTAGCTCCTCCTCTT
231 SEQ ID NO: 631 -24.1 -38.9 89.5 -14.8 0 -6 TTCACAGTAGCTCCTCCTCT
232 SEQ ID NO: 632 -24.1 -38.9 89.5 -14.8 0 -5.7 AAGTCTTCGCTGCTTGGAGT
296 SEQ ID NO: 633 -24.1 -37.4 88 -11.7 -1.6 -10.2
GTCGACCTATTGAGGTTTGC 511 SEQ ID NO: 634 -24.1 -35.6 85.3 -6.8 -4.7 -13.8
TGGATGCTGAACTCTTGGGG 546 SEQ ID NO: 635 -24.1 -39.5 90 -15.4 0 -6.9
CAGTCTGGCCCTTCAAATGT 636 SEQ ID NO: 636 -24.1 -36.9 87.3 -12.3 0 -7.6
AATTTCACATTGCCACCGTT 661 SEQ ID NO: 637 -24.1 -32.4 81.5 -8.3 0 -0.9
AAAGGAATGAATCGTCTTCT 822 SEQ ID NO: 638 -24.1 -30.9 77.8 -6 -0.6 -8.4
GAGTTCGATGAAATCTTCTT 971 SEQ ID NO: 639 -24.1 -30 76 -5.9 0 -7.4
ACTGTGCCCAGTTTCTCTTG 1012 SEQ ID NO:640 -24.1 -36.8 87.5 -11.1 -1.4 -10.3
AAAGCTTGCCTGACAGTAAA 1031 SEQ ID NO:64l -24.1 -33.3 83.8 -8.6 -0.3 -6.9
ACTCCTGTAGTGGCCTGCTG 1633 SEQ ID NO: 642 -24.1 -41.8 93.9 -15.4 -2.2 -11.7
AGCGTAGTCATGATCCTCCA 1801 SEQ ID NO: 643 -24.1 -38.4 87.9 -13.5 0 -9.4
CTTCCCTCTTGACAATGGCT 2229 SEQ ID NO: 644 -24.1 -37.1 87.2 -13 0 -4.1
TGGAGATTACGTCCACATAT 2729 SEQ ID NO: 645 -24.1 -33.9 82.4 -7.7 -2.1 -7.2
TGCCATCCTTCTTTGACTGT 2748 SEQ ID NO: 646 -24.1 -36 85.7 -11.9 0 -1
AATTCTCCACTGAAGCAGAT 3253 SEQ ID NO: 647 -24.1 -34 82.2 -9.2 -0.5 -7.2
CACAGTTTACCCAGCAGGTC 3326 SEQ ID NO: 648 -24.1 -38.2 89.7 -13.2 -0.7 -6.4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GTTGGAATGAGAAGGGTGGT
4370 SEQ ID NO: 649 -24.1 -37.2 87.3 -13.1 0 -0.2
TTGTCATTCAACTGCTGGGG
4533 SEQ ID NO: 50 -24.1 -36.9 87.2 -12.8 0 -3.4
GCAGTTCCCGCCGCAGCCGA
66 SEQ ID NO: 651 -24 -46.2 98.5 -20.5 -1.7 -7.5
GAAGACGCAGAAACCTTCAC
247 SEQ ID NO: 652 -24 -35.3 84.7 -9.2 -2.1 -6.8
TTTTCTGTTTTCACTTGGGG
952 SEQ ID NO: 653 -24 -31.8 81.4 -7.8 0 -6.4
GCAGACATTTTATTACCAAT
1066 SEQ ID NO: 654 -24 -28.2 75.3 -4.2 0 -2.7
ATCTGGTCTCATGCTGGGGC
1316 SEQ ID NO: 655 -24 -42.2 92.4 -14.2 -3.8 -15.3
TTAAGACTCCATAATGACAT
1422 SEQ ID NO: 656 -24 -29.9 76.8 -5.9 0 -3.5
TCCTTCCACTGCTCTTTTGA
1466 SEQ ID NO: 657 -24 -36 85.9 -12 0 -1.7
TCGATAGCGGCATGCTGGGC
1550 SEQ ID NO: 658 -24 -43 94 -14.1 -2.6 -17.9
GTAAGGTTTTCATACAGAGA
2121 SEQ ID NO: 659 -24 -30.8 79.1 -6.8 0 -5.1
TTGTCAGTTGATAAAACCGC
2271 SEQ ID NO: 660 -24 -31.7 80.6 -5.3 -2.2 -12.2
AACCACATGTAGTGCGTATT
2595 SEQ ID NO: 661 -24 -33.1 82.9 -6.7 -2.4 -10.2
AAACCACATGTAGTGCGTAT
2596 SEQ ID NO: 662 -24 -33.1 82.9 -6.7 -2.4 -10.2
TTAAGGTTTCTAATTTCTGG
2710 SEQ ID NO: 663 -24 -27.9 75.6 -3.9 0 -4
ACTTTTTTTCAGGTTTCCAT
2908 SEQ ID NO: 664 -24 -28.4 75.9 -4.4 0 -5.3
GAATACTGGAGATTTGAGTC
3193 SEQ ID NO: 665 -24 -32.3 79.7 -7.8 0 -8
GTCACTGGACTAGGTGCTCT
3309 SEQ ID NO: 666 -24 -40 90.8 -13.4 -2.6 -11.3
AGGAATTACTTTGTCTGATT
3544 SEQ ID NO: 667 -24 -29.3 76.3 -5.3 0 -3.6
GCCATATTAGAGATTTAGTT
3584 SEQ ID NO: 668 -24 -28.8 76.1 -4.8 0 -6.1
TGCTGGGGGAGGGCTGTATG
4521 SEQ ID NO: 669 -24 -44.2 96.5 -19 -1.1 -5
CTTGTTGTAGGATAGAAAGG
4647 SEQ ID NO: 670 -24 -31.6 80.8 -7.1 0 -8
GAGTCCATCAGTGAATATCA
121 SEQ ID NO: 671 -23.9 -33.7 80.9 -9.8 0 -6.7
AGTCTTCGCTGCTTGGAGTC
295 SEQ ID NO: 672 -23.9 -38.9 89.1 -13.2 -1.8 -10.2
TGGGGTTCTCTGGAACACTG
531 SEQ ID NO: 673 -23.9 -39.4 90.4 -11.6 -3.9 -13.5
GTGCCAGTCTGGCCCTTCAA
640 SEQ ID NO: 674 -23.9 -41.6 93.3 -11.8 -3.7 -20 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecul Site oligo binding formation Duplex structure oligo oligo
CGTCTTCTCCCGCCAGAGGA
810 SEQ ID NO: 675 -23.9 -42.9 93.7 -15 -4 -13.1
AATCAGATCAGGAGCAAAAC
1997 SEQ ID NO: 676 -23.9 -32.8 80.6 -8.9 0 -5.1
TTCATACAGAGATACTCTTC
2113 SEQ ID NO: 677 -23.9 -31.5 78.6 -6.6 -0.9 -8
CGCTGCCAGTTCTGGCTGGA
2254 SEQ ID NO: 678 -23.9 -43.1 94.9 -12.8 -6.1 -20.6
TAAGGAGATTTTCAACCACT
2313 SEQ ID NO: 679 -23.9 -31.5 80.1 -6.7 0 -9.8
CTAACTATACAGGGGGGGGA
2867 SEQ ID NO: 680 -23.9 -40.1 92.8 -14.6 -1.6 -6.8
CAGTAATAGCTATAAAAGGC
2959 SEQ ID NO: 681 -23.9 -29.7 78.9 -4.7 -0.8 -9.5
GGGCCTCTTGGTAGTTATTT
3374 SEQ ID NO: 682 -23.9 -35 86.1 -8.3 -2.8 -9.3
AGCCTCAGCAACCTTCACTG
4302 SEQ ID NO: 683 -23.9 -39.2 90.3 -14.4 -0.7 -3.8
ATGAGAAGGGTGGTCAGAAT
4364 SEQ ID NO: 684 -23.9 -36.4 85.4 -12.5 0 -5.5
TATTTGGAAATAAACTCTTG
4663 SEQ ID NO: 685 -23.9 -25.7 71.6 -1.8 0 -3.4
AGTCTGATTGAGAAGCGACA
279 SEQ ID NO: 686 -23.8 -35.7 84.2 -10.6 -1.2 -5.6
CCCATTGAGAGTGAAACTGC
373 SEQ ID NO: 687 -23.8 -36 85.5 -12.2 0 -5.1
CTGAAGATACATCAGAGTGA
609 SEQ ID NO: 688 -23.8 -33.5 80.9 -7.6 -2.1 -7.2
TTTTGGGTTTAGTGTCCGGT
882 SEQ ID NO: 689 -23.8 -34.8 86.3 -7.3 -3.7 -7.4
CCCAGGGGTGCAGAGTTCGA
983 SEQ ID NO: 690 -23.8 -43.3 95 -17.9 -1.2 -11
GTCATCCAGGTGTAAGTTCC
1886 SEQ ID NO: 691 -23.8 -36.5 86.3 -12.1 -0.3 -6.2
TTACATTGGTCGTACATGCA
2044 SEQ ID NO: 692 -23.8 -33.3 82.7 -9.5 0 -6.1
TGGAGATTTGAGTCAATTTT
3187 SEQ ID NO: 693 -23.8 -29.1 75.4 -3.7 -0.7 -11
AGATTTAGTTTTTGGGTAAA
3574 SEQ ID NO: 694 -23.8 -26.5 73.8 -2.7 0 -3.6
TAACCAATTGGTGACAGATG
3635 SEQ ID NO: 695 -23.8 -32.7 81.4 -5.1 -3.2 -15.6
GGCACACATTAGGGATGTGT
3738 SEQ ID NO: 696 -23.8 -37.4 87.7 -9 -4.6 -16.3
GTTGGTAAGGTGCACACAGA
3837 SEQ ID NO: 697 -23.8 -37.1 88.3 -11.8 0 -11
AGACTCACTGTTGGAATGAG
4379 SEQ ID NO: 698 -23.8 -35.1 83.8 -10.4 -0.8 -7.8
GTTGTAGGATAGAAAGGAAT
4644 SEQ ID NO: 699 -23.8 -30.9 79 -7.1 0 -3.8
GCAGAAACCTTCACAGTAGC
241 SEQ ID NO: 700 -23.7 -36.1 86.6 -12.4 0 -2.7 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TGATTGAGAAGCGACAGCCA
275 SEQ ID NO: 701 -23.7 -37.8 87.4 -13 -1 -4.6
GCGCATTGCTTACTGAGCCT
333 SEQ ID NO: 702 -23.7 -38.7 89.9 -13.2 -1.4 -11.4
CCTATTGAGGTTTGCAATGC
506 SEQ ID NO: 703 -23.7 -33.6 83.1 -8.8 -1 -9.2
ACACTGGTCGACCTATTGAG
517 SEQ ID NO: 704 -23.7 -36.8 86.7 -8.8 -1 -16.7
GAACTCTTGGGGTTCTCTGG
538 SEQ ID NO: 705 -23.7 -38.4 88.8 -9.2 -4.9 -19
AAGATACATCAGAGTGAGTT
606 SEQ ID NO: 706 -23.7 -32.1 79.6 -8.4 0 -3.5
CCGTTGGTGCCAGTCTGGCC
646 SEQ ID NO: 707 -23.7 -44.1 96.3 -14.5 -3.7 -20
GGTCTGTGGTATACAATTTC
675 SEQ ID NO: 708 -23.7 -32 80.7 -8.3 0 -7.1
GAGTTTGGGAGGTGGTCCTG
1373 SEQ ID NO: 709 -23.7 -40.6 91.7 -13.8 -3.1 -7.6
GCTTCATCAGAGCACACCAG
1396 SEQ ID NO: 710 -23.7 -38.5 88.1 -11.7 -3.1 -9
GCCTGAAGACATTTTCGATA
1564 SEQ ID NO: 711 -23.7 -32.3 80.2 -7.9 -0.4 -8.2
TCCAGCCTGAAGACATTTTC
1568 SEQ ID NO: 712 -23.7 -35 84 -11.3 0 -6.8
TGTAAGTTCCTGAAACCTGG
1876 SEQ ID NO: 713 -23.7 -34.7 85.3 -11 0 -3.1
AGTAAGGTTTTCATACAGAG
2122 SEQ ID NO: 714 -23.7 -30.5 79.1 -6.8 0 -5.1
CCTCTTGACAATGGCTTTTC
2225 SEQ ID NO: 715 -23.7 -33.5 82.5 -9.8 0 -3.8
TTTGGGCACTGGTGGTTTAG
2769 SEQ ID NO: 716 -23.7 -36.6 88.7 -10.9 -0.5 -12.2
TTTCATCCAGCCAACTGTGA
2823 SEQ ID NO: 717 -23.7 -36 85.6 -11.1 -1.1 -5.1
CAGGACTTGTGTTAAACTCT
3760 SEQ ID NO: 718 -23.7 -32.4 81.9 -8 -0.5 -5.9
TGTAATCTCACTGGGTCAGA
4321 SEQ ID NO: 719 -23.7 -36.7 86.2 -11.8 -0.1 -10.4
TGACAGACTCACTGTTGGAA
4383 SEQ ID NO: 720 -23.7 -36.2 85.7 -8.9 -3.6 -9.2
TTACATACTTTAGTGCAAGG
4429 SEQ ID NO: 721 -23.7 -30.4 80 -6.7 0 -6
ACCTTCACAGTAGCTCCTCC
235 SEQ ID NO: 722 -23.6 -39.9 91 -16.3 0 -6.1
AAAGTCTTCGCTGCTTGGAG
297 SEQ ID NO: 723 -23.6 -36.1 86.3 -11.2 -0.9 -10.2
TCGACCTATTGAGGTTTGCA
510 SEQ ID NO: 724 -23.6 -35.5 85.4 -8.1 -3.8 -13.8
AATGTCAAAGGTGCTTTGGT
692 SEQ ID NO: 725 -23.6 -33.1 82.8 -7.4 -2.1 -9.6
GGAGGAGAGCTTACATCTGG
1330 SEQ ID NO: 726 -23.6 -39 89 -13.9 0.1 -10.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GTTGAGCGTAGTCATGATCC
1805 SEQ ID NO: 727 -23.6 -36.1 84.7 -11.7 0 -9.4
CATCCAGGTGTAAGTTCCTG
1884 SEQ ID NO: 728 -23.6 -36.1 86.3 -8.8 -3.7 -9.9
TACATGCAGGGTAGAGTCAT
2032 SEQ ID NO: 729 -23.6 -36.8 86.9 -12.5 -0.2 -8.7
ACAGAGATACTCTTCATAAG
2108 SEQ ID NO: 730 -23.6 -31.2 78.6 -6.6 -0.9 -8
TGGCTGGAGTTTCCTTCCCT
2242 SEQ ID NO: 731 -23.6 -40.5 92.3 -13.2 -3.6 -14.4
CTGGCTGGAGTTTCCTTCCC
2243 SEQ ID NO: 732 -23.6 -40.5 91.8 -13.2 -3.6 -14.4
GTTAGGACTGTTAATTTGCA
3283 SEQ ID NO: 733 -23.6 -30.3 79.3 -6.7 0 -5.8
AGTTAGGACTGTTAATTTGC
3284 SEQ ID NO: 734 -23.6 -30.3 79.3 -6.7 0 -6.8
TTTTTGGGTAAAGTTTAGTG
3566 SEQ ID NO: 735 -23.6 -27.3 76 -3.7 0 -3.8
ACTTGTGTTAAACTCTATGG
3756 SEQ ID NO: 736 -23.6 -30.3 79.2 -6.7 0 -3.4
TTTGGCCACCTTGAATAGAA
3908 SEQ ID NO: 737 -23.6 -33.6 83.6 -8.4 0 -11.2
GGGGAGGGCTGTATGTGAAA
4516 SEQ ID NO: 738 -23.6 -39.7 91.1 -16.1 0 -4.9
CCTTCACAGTAGCTCCTCCT
234 SEQ ID NO: 739 -23.5 -39.8 91 -16.3 0 -6
ACCCCAGGGGTGCAGAGTTC
985 SEQ ID NO: 740 -23.5 -44 96.2 -14.9 -3.3 -19.4
TTGAGGAGCTGGATGGAGGA
1344 SEQ ID NO: 741 -23.5 -41.2 91.4 -17.7 0 -5.3
GTCCTTCCACTGCTCTTTTG
1467 SEQ ID NO: 742 -23.5 -35.8 85.8 -12.3 0 -1.7
CATTCCAGCCTGAAGACATT
1571 SEQ ID NO: 743 -23.5 -34.9 83.8 -11.4 0 -6.5
TATTGCCTTTGCCCATTTCA
1856 SEQ ID NO: 744 -23.5 -33.5 83.4 -10 0 -2
GGTCATCCAGGTGTAAGTTC
1887 SEQ ID NO: 745 -23.5 -36.5 86.3 -12 -0.9 -6.2
TGCTCATTAATAATCAGATC
2008 SEQ ID NO: 746 -23.5 -29.1 74.9 -5.6 0 -4.2
AGGTTTTCATACAGAGATAC
2118 SEQ ID NO: 747 -23.5 -31 78.9 -6.8 -0.5 -3.9
TCTGGCTGGAGTTTCCTTCC
2244 SEQ ID NO: 748 -23.5 -39.6 90.4 -13.2 -2.8 -13.1
AAGGAGATTTTCAACCACTT
2312 SEQ ID NO: 749 -23.5 -31.1 79.4 -6.7 0 -9.8
ATAAACCACATGTAGTGCGT
2598 SEQ ID NO: 750 -23.5 -33.1 82.9 -7.9 -1.7 -9.4
CTTGGCCCTCTATAAACCAC
2609 SEQ ID NO: 751 -23.5 -36.2 87.4 -12.2 -0.1 -7.5
CCTTCTTTGACTGTGGAGAT
2742 SEQ ID NO: 752 -23.5 -35 83.9 -9.6 -1.9 -8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TCTTTTGCAACCATCATCCA
3344 SEQ ID NO: 753 -23.5 -33.7 82.3 -10.2 0 -6.2
AAAGTCTCTCAGCTGTGTTA
3523 SEQ ID NO: 754 -23.5 -33.9 83.4 -9.7 0 -9.1
TAAAAGTCTCTCAGCTGTGT
3525 SEQ ID NO: 755 -23.5 -33.9 83.4 -9.7 0 -9.1
CATATTAGAGATTTAGTTTT
3582 SEQ ID NO: 756 -23.5 -23.9 68.4 0.4 0 -6.3
GAGGCAGAGGATAACTTCCT
4342 SEQ ID NO: 757 -23.5 -37.8 87.8 -11.1 -3.2 -8.8
TAATTTCTCCAAAATACTGA
4739 SEQ ID NO: 758 -23.5 -27.2 73.4 -3.7 0 -0.4
GCTGCGCATTGCTTACTGAG
336 SEQ ID NO: 759 -23.4 -37.5 88.1 -12.2 -1 -11.9
CTGGCTGCTGCGCATTGCTT
342 SEQ ID NO: 760 -23.4 -40.4 92 -13.1 -1.8 -16
AGTCCCATTGAGAGTGAAAC
376 SEQ ID NO: 761 -23.4 -35.1 84 -11.7 0 -4.5
CCCATCACTTTTGTTTCTGT
409 SEQ ID NO: 762 -23.4 -32.1 81 -8.7 0 0
CAGCAGACACAGCAGTGGAT
561 SEQ ID NO: 763 -23.4 -39.5 89.7 -14.1 -2 -9.1
TGGTCTGTGGTATACAATTT
676 SEQ ID NO: 764 -23.4 -31.7 80.7 -8.3 0 -7.1
TTGGTCTGTGGTATACAATT
677 SEQ ID NO: 765 -23.4 -31.7 80.7 -8.3 0 -7.1
TTTGGTCTGTGGTATACAAT
678 SEQ ID NO: 766 -23.4 -31.7 80.7 -8.3 0 -7.1
AAGGAATGAATCGTCTTCTC
821 SEQ ID NO: 767 -23.4 -32.4 79.4 -8.1 -0.7 -9.2
CAAAAGGAATGAATCGTCTT
824 SEQ ID NO: 768 -23.4 -29.4 76 -6 0 -6.8
CTTGCCTGACAGTAAACTGT
1027 SEQ ID NO: 69 -23.4 -34.6 85.2 -8.6 -2.6 -10.1
GTACATCTGTCCTCCAGAGG
1109 SEQ ID NO: 770 -23.4 -39.1 88.9 -13.4 -2.3 -11.1
ACGGGAATTGGTGGAATGAC
1189 SEQ ID NO: 771 -23.4 -36.5 86 -13.1 0 -3.8
GAGCTTACATCTGGTCTCAT
1324 SEQ ID NO: 772 -23.4 -35.7 84.3 -9.5 -2.8 -8.8
ATTTTCGATAGCGGCATGCT
1554 SEQ ID NO: 773 -23.4 -34.7 83.9 -9.5 -0.7 -11.8
CTGGTATTGCCTTTGCCCAT
1860 SEQ ID NO: 774 -23.4 -36.9 88 -10.7 -2.8 -10.7
ACATGCAGGGTAGAGTCATT
2031 SEQ ID NO: 775 -23.4 -36.4 86.2 -12.5 -0.2 -6.2
TGTTTACATTGGTCGTACAT
2047 SEQ ID NO: 776 -23.4 -30.9 79.5 -7.5 0 -6.1
ATGTGTTTACATTGGTCGTA
2050 SEQ ID NO: 777 -23.4 -30.9 79.5 -7.5 0 -7.3
GAGTCAATTTTTGTGGTCTT
3178 SEQ ID NO: 778 -23.4 -30 77.6 -6.6 0 -5.2 -- -
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
TTGAGTCAATTTTTGTGGTC
3180 SEQ ID NO: 779 -23.4 -30 77.5 -6.6 0 -7.3
TTTTTTGACAAGAATACTGG
3204 SEQ ID NO: 780 -23.4 -27.6 74.7 -3.7 0 -7.8
ACTTTGTCTGATTAAAAGTC
3537 SEQ ID NO: 781 -23.4 -28 74.8 -3.7 -0.7 -6.9
GAGATTTAGTTTTTGGGTAA
3575 SEQ ID NO: 782 -23.4 -28 75.8 -4.6 0 -3.7
TGGCACACATTAGGGATGTG
3739 SEQ ID NO: 783 -23.4 -37.3 87.6 -11.2 -2.6 -12.8
TCTGAGACCATCGCTGCCTG
4230 SEQ ID NO: 784 -23.4 -41.4 91.4 -18 0 -5.3
CCTCAGCAACCTTCACTGCA
4300 SEQ ID NO: 785 -23.4 -39.2 90.3 -13.7 -2.1 -5.7
ACTGGTCGACCTATTGAGGT
515 SEQ ID NO: 786 -23.3 -38 88.5 -10.5 -2.5 -16.5
TACCAATTATATTTGCTCCA
1053 SEQ ID NO: 787 -23.3 -29.7 77.8 -6.4 0 -4.6
AATGGCAGACATTTTATTAC
1070 SEQ ID NO: 788 -23.3 -28.2 75.3 -4.2 -0.5 -3.6
TGAAGCTTCATCAGAGCACA
1400 SEQ ID NO: 789 -23.3 -36.3 85.3 -9.5 -3.3 -14.5
TCCAGCACATAGGTAATTGT
1490 SEQ ID NO: 790 -23.3 -34.1 84 -10.8 0 -2.7
TGAATATTTTGGTATCTGAT
2408 SEQ ID NO: 791 -23.3 -27.6 73.1 -4.3 0 -5.4
CTCTATAAACCACATGTAGT
2602 SEQ ID NO: 792 -23.3 -30.9 79.7 -6.6 -0.9 -7.4
CCAAGTCTTGGCCCTCTATA
2615 SEQ ID NO: 793 -23.3 -37.6 88.8 -11.3 -0.5 -14.2
GGAGATTACGTCCACATATT
2728 SEQ ID NO: 794 -23.3 -32.7 80.9 -7.7 -1.7 -6.4
TCCTTCTTTGACTGTGGAGA
2743 SEQ ID NO: 795 -23.3 -36.3 85.7 -11.1 -1.9 -6.9
AACTTTTTTTCAGGTTTCCA
2909 SEQ ID NO: 796 -23.3 -28.2 76.1 -4.4 -0.1 -5.3
CAGGGCCTCTTGGTAGTTAT
3376 SEQ ID NO: 797 -23.3 -37.4 89 -11.3 -2.8 -10.7
ATTACTTTGTCTGATTAAAA
3540 SEQ ID NO: 798 -23.3 -24.6 69.5 -1.2 0 -3.4
AATTACTTTGTCTGATTAAA
3541 SEQ ID NO: 799 -23.3 -24.6 69.5 -1.2 0 -3.4
TATTAGAGATTTAGTTTTTG
3580 SEQ ID NO: 800 -23.3 -23.7 68.5 0.4 0 -7.1
CCAGATTAGTGAATACCAAT
4022 SEQ ID NO: 801 -23.3 -30.8 78.7 -7.5 0 -3
TCCCCTAGAGCAAACTGTTT
4254 SEQ ID NO: 802 -23.3 -35.9 87.1 -11.9 -0.4 -5.1
TGTCATTCAACTGCTGGGGG
4532 SEQ ID NO: 803 -23.3 -39.3 90.2 -16 0 -7.3
AATAAACTCTTGTTGTAGGA
4655 SEQ ID NO: 804 -23.3 -29.3 77.4 -6 0 -4.7 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
GGATGCTGAACTCTTGGGGT
545 SEQ ID NO: 805 -23.2 -39.6 90.1 -15.4 -0.7 -9.6
AAAAGGAATGAATCGTCTTC
823 SEQ ID NO: 806 -23.2 -29.7 76.1 -6 0 -7.8
TTCGATGAAATCTTCTTTTT
968 SEQ ID NO: 807 -23.2 -26 70.4 -2.8 0 -4.6
GACAGTAAACTGTGCCCAGT
1020 SEQ ID NO: 808 -23.2 -37.1 88.3 -10.6 -3.3 -12.1
CAGGAAAGCTTGCCTGACAG
1035 SEQ ID NO: 809 -23.2 -37.9 88.7 -8.6 -4.3 -20.3
GGGGCTTGAATAGCCATTAG
1301 SEQ ID NO: 810 -23.2 -36.3 87.1 -7.4 -5.7 -14.1
AGAGCTTACATCTGGTCTCA
1325 SEQ ID NO: 811 -23.2 -36.7 86.2 -10.5 -3 -8.8
GCACACCAGGCAGAGTTTGG
1385 SEQ ID NO: 812 -23.2 -40.2 91.7 -13.7 -3.3 -12.4
TTCCTGAAACCTGGTATTGC
1870 SEQ ID NO: 813 -23.2 -34.9 85.1 -11 -0.4 -8
CCCCAGAGCAAATGCCATAA
1940 SEQ ID NO: 814 -23.2 -37.1 87.9 -13.1 -0.6 -6.1
CACCCCAGAGCAAATGCCAT
1942 SEQ ID NO: 815 -23.2 -39.2 90 -15.2 -0.6 -6.1
TGTAGGTCATTCTAATTTCA
2193 SEQ ID NO: 816 -23.2 -297 77 -5.5 -0.9 -4.7
TGATGACGACTCAACTGCTT
2641 SEQ ID NO: 817 -23.2 -35.5 84.3 -12.3 0 -2.6
AACTATACAGGGGGGGGATA
2865 SEQ ID NO: 818 -23.2 -39.1 91.3 -15.9 0 -1.6
ATCATCCACAGTTTACCCAG
3332 SEQ ID NO: 819 -23.2 -35.1 84.6 -11.9 0 -1.2
AGCTTTCTTGCCATATAGCC
3424 SEQ ID NO: 820 -23.2 -35.2 85.6 -10.9 -1 -7.3
GTAAAGTTTAGTGAGAGGAA
3559 SEQ ID NO: 821 -23.2 -30.6 79.4 -7.4 0 -3.8
CCACCTTGAATAGAAATCAA
3903 SEQ ID NO: 822 -23.2 -30.4 78.1 -7.2 0 -1.6
ACCATCGCTGCCTGTATGAA
4224 SEQ ID NO: 823 -23.2 -37.9 88.2 -13.8 -0.7 -5.7
TACATACTTTAGTGCAAGGG
4428 SEQ ID NO: 824 -23.2 -32.8 83.4 -9.1 -0.1 -8
GCTGGTACCTCTATGCAAAC
4578 SEQ ID NO: 825 -23.2 -36.4 87.2 -10.8 -2.4 -10.6
TGGTCGACCTATTGAGGTTT
513 SEQ ID NO: 826 -23.1 -35.5 85.5 -8.1 -3.5 -16.5
TTCATCTATCAACAGGTCTG
776 SEQ ID NO: 827 -23.1 -33.2 81.1 -9.3 0 -9.4
AGAGTTCGATGAAATCTTCT
972 SEQ ID NO: 828 -23.1 -31.2 77.6 -7.5 0 -8.4
CATCTGTCCTCCAGAGGTAC
1106 SEQ ID NO: 829 -23.1 -39.1 88.9 -13.4 -2.6 -12
CCAGCCTGAAGACATTTTCG
1567 SEQ ID NO: 830 -23.1 -35 84.4 -11.3 -0.3 -6.8 ., .
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CATATAACACTTCAGGTTCA
1749 SEQ ID NO: 831 -23.1 -30.7 78.8 -7.6 0 -4.5
GTAAGTTCCTGAAACCTGGT
1875 SEQ ID NO: 832 -23.1 -34.8 85.4 -11 -0.4 -6.1
TGCCATAAGAAACATCCAGG
1928 SEQ ID NO: 833 -23.1 -35 84.6 -11.9 0 -1.6
GCTGCCAGTTCTGGCTGGAG
2253 SEQ ID NO: 834 -23.1 -42.8 94.2 -13.2 -6.3 -20.6
TTGAATATTTTGGTATCTGA
2409 SEQ ID NO: 835 -23.1 -27.4 73.2 -4.3 0 -5.4
ACCACATGTAGTGCGTATTT
2594 SEQ ID NO: 836 -23.1 -33.1 82.9 -7.9 -2.1 -9.9
CTGTGGAGATTACGTCCACA
2732 SEQ ID NO: 837 -23.1 -36.8 86.6 -7.7 -6 -15
CTTTGACTGTGGAGATTACG
2738 SEQ ID NO: 838 -23.1 -33.1 81.9 -9.4 -0.3 -5.9
AACAGCTGTACAATAACTTG
3050 SEQ ID NO: 839 -23.1 -30.1 79.2 -4.3 0 -13.6
AGTTTATTTGGGAAAGCTAC
3089 SEQ ID NO: 840 -23.1 -30.2 79.4 -5.8 -1.2 -9
TTAGGACTGTTAATTTGCAC
3282 SEQ ID NO: 841 -23.1 -30.3 79.3 -6.7 -0.1 -5.8
AAAAGTCTCTCAGCTGTGTT
3524 SEQ ID NO: 842 -23.1 -33.5 82.7 -9.7 0 -9.1
TGCCATATTAGAGATTTAGT
3585 SEQ ID NO: 843 -23.1 -30 77.7 -6.9 0 -5.4
GCACACATTAGGGATGTGTA
3737 SEQ ID NO: 844 -23.1 -35.4 85.4 -6.9 -5.3 -18
ATGGCACACATTAGGGATGT
3740 SEQ ID NO: 845 -23.1 -36.3 86.1 -12.6 -0.3 -7.7
ACAGAAAGTTGGTAAGGTGC
3844 SEQ ID NO: 846 -23.1 -34.6 85.4 -11.5 0 -3.2
TGCCTTTAAGGATGTAAGCA
4477 SEQ ID NO: 847 -23.1 -33.8 84.3 -9.7 -0.7 -9.3
ATTTCTCCAAAATACTGAAA
4737 SEQ ID NO: 848 -23.1 -26.8 72.6 -3.7 0 -1.9
AATTTCTCCAAAATACTGAA
4738 SEQ ID NO: 849 -23.1 -26.8 72.6 -3.7 0 -0.4
ACAGTAGCTCCTCCTCTTAG
229 SEQ ID NO: 850 -23 -37.8 88.8 -14.8 0 -6.1
CACAGTAGCTCCTCCTCTTA
230 SEQ ID NO: 851 -23 -37.8 88.7 -14.8 0 -6
TTCCCATCACTTTTGTTTCT
411 SEQ ID NO: 852 -23 -31.1 79.3 -8.1 0 0
GTGGATGCTGAACTCTTGGG
547 SEQ ID NO: 853 -23 -38.4 88.4 -15.4 0 -4.5
TCACATTGCCACCGTTGGTG
657 SEQ ID NO: 854 -23 -38.3 89.2 -12.4 -2.9 -12.8
TGCAAAATGTCAAAGGTGCT
697 SEQ ID NO: 855 -23 -33.1 82.7 -9.4 -0.3 -8.2
ACAGTAAACTGTGCCCAGTT
1019 SEQ ID NO: 856 -23 -35.6 87.1 -9.7 -2.9 -12.5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecul Site oligo binding formation Duplex structure oligo oligo
CGGGAATTGGTGGAATGACA
1188 SEQ ID NO: 857 -23 -36.4 85.9 -13.4 0 -3.3
TAATCAGATCAGGAGCAAAA
1998 SEQ ID NO: 858 -23 -31.9 79.6 -8.9 0 -5.3
GCTCATTAATAATCAGATCA
2007 SEQ ID NO: 859 -23 -29.1 74.9 -6.1 0 -5.2
GAGATTACGTCCACATATTA
2727 SEQ ID NO: 860 -23 -30.7 78.2 -7.7 0 -3
AAAAGTATGAAGAGAAAGTT
2801 SEQ ID NO: 861 -23 -26.7 72.8 -3.7 0 -1
AAAAAGTATGAAGAGAAAGT
2802 SEQ ID NO: 862 -23 -26.7 72.8 -3.7 0 -0.7
CAGACAGTAATAGCTATAAA
2963 SEQ ID NO: 863 -23 -28.8 76.9 -4.9 0 -9.8
GTTTATTTGGGAAAGCTACG
3088 SEQ ID NO: 864 -23 -30.5 79.9 -7 -0.1 -6.8
GACAAGAATACTGGAGATTT
3198 SEQ ID NO: 865 -23 -30.8 78.1 -7.8 0 -2.8
CTGGTACCTCTATGCAAACT
4577 SEQ ID NO: 866 -23 -35.1 85.7 -10.8 0 -10.6
AGAGGAGCCGCTCGCCCGCC
93 SEQ ID NO: 867 -22.9 -48.8 100.9 -22.3 -3.4 -14.6
CTGCAAAATGTCAAAGGTGC
698 SEQ ID NO: 868 -22.9 -33.1 82.6 -10.2 0 -5.8
TACATCTGTCCTCCAGAGGT
1108 SEQ ID NO: 869 -22.9 -39.1 89.2 -13.4 -2.8 -11.8
AGCCTGAAGACATTTTCGAT
1565 SEQ ID NO: 870 -22.9 -33.1 81.1 -9.5 -0.4 -8.2
GGTGAGTTGTGGTAACGTTG
1700 SEQ ID NO: 871 -22.9 -35 85.7 -12.1 0 -5.9
TAACATGTTGAGCGTAGTCA
1811 SEQ ID NO: 872 -22.9 -33.4 82.8 -9.5 0 -10
CAGAGATACTCTTCATAAGA
2107 SEQ ID NO: 873 -22.9 -31.4 78.5 -7.6 -0.8 -7.9
TACAGAGATACTCTTCATAA
2109 SEQ ID NO: 874 -22.9 -30.4 77.7 -6.6 -0.8 -7.9
TTTCCATTTGAATATTTTGG
2416 SEQ ID NO: 875 -22.9 -25.4 70.6 -2.5 0 -6.4
GCTCTATACCAGTTAGGACT
3294 SEQ ID NO: 876 -22.9 -35.8 86.4 -11.5 -1.3 -6
AACCAATTGGTGACAGATGG
3634 SEQ ID NO: 877 -22.9 -34.7 83.9 -5.1 -6.7 -15.6
AGGACTTGTGTTAAACTCTA
3759 SEQ ID NO: 878 -22.9 -31.6 81.1 -8 -0.5 -3.7
GGCTTAGTAAATATGTTAAG
3874 SEQ ID NO: 879 -22.9 -27.3 75.4 -4.4 0 -5.1
GACAGACTCACTGTTGGAAT
4382 SEQ ID NO: 880 -22.9 -35.2 83.9 -8.9 -3.4 -9.2
GCTGGGGGAGGGCTGTATGT
4520 SEQ ID NO:881 -22.9 -44.3 96.6 -20.2 -1.1 -5
TCTGAAGCTTCTGTTGTCAT
4546 SEQ ID NO: 882 -22.9 -33.9 82.3 -8.8 -1.3 -12.5 .
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
TAGGATAGAAAGGAATTAGT
4640 SEQ ID NO: 883 -22.9 -30 78 -7.1 0 -0.8 GTAGGATAGAAAGGAATTAG
4641 SEQ ID NO: 884 -22.9 -30 78 -7.1 0 -0.5 TGTAGGATAGAAAGGAATTA
4642 SEQ ID NO: 885 -22.9 -30 77.9 -7.1 0 -0.5 TTTCTCCAAAATACTGAAAA
4736 SEQ ID NO: 886 -22.9 -26.6 72.8 -3.7 0 -2.9
TACAGTCCCATTGAGAGTGA 379 SEQ ID NO: 887 -22.8 -36.7 86.2 -12.2 -1.6 -10.8
CTATTGAGGTTTGCAATGCT 505 SEQ ID NO: 888 -22.8 -32.4 81.5 -8.8 -0.6 -7.7
CTGGTCGACCTATTGAGGTT 514 SEQ ID NO: 889 -22.8 -36.7 86.9 -9.6 -3.7 -16.5
TTGCTTAATTACCCCAGGGG 995 SEQ ID NO: 890 -22.8 -37 89.4 -10.5 -2.4 -15.6
ATTACCAATTATATTTGCTC 1055 SEQ ID NO: 891 -22.8 -26.3 72.3 -3.5 0 -4.3
TCCTCCAGAGGTACTCACAC 1100 SEQ ID NO: 892 -22.8 -40.2 90.8 -15.4 -1.7 -11.8
CAACGGGAATTGGTGGAATG 1191 SEQ ID NO: 893 -22.8 -34.9 84.4 -12.1 0 -3.2
GGTCCTGTTGTTGCTGTTGA 1360 SEQ ID NO: 894 -22.8 -36.9 87.5 -14.1 0 -5.6
CTGTAGTGGCCTGCTGAATT 1629 SEQ ID NO: 895 -22.8 -37.1 88 -11.5 -2.2 -13.6
AACATGTTGAGCGTAGTCAT 1810 SEQ ID NO: 96 -22.8 -33.2 81.9 -9.5 0 -9.8
ATCAGATCAGGAGCAAAACA 1996 SEQ ID NO: 897 -22.8 -34 82.1 -11.2 0 -5.3
ATTTCATCCAGCCAACTGTG 2824 SEQ ID NO: 898 -22.8 -34.7 83.8 -11.1 -0.6 -3.9
CTGAAGCAGATATATACAAA 3244 SEQ ID NO: 899 -22.8 -28.8 75.9 -6 0 -6.7
CTGTTAATTTGCACAACCTA 3276 SEQ ID NO: 900 -22.8 -30 79.2 -6.7 -0.1 -5.4
AGTTGGTAAGGTGCACACAG 3838 SEQ ID NO: 901 -22.8 -36.8 88.4 -12.5 0 -11
ATATGTTAAGTTTTGAGTTT
3864 SEQ ID NO: 902 -22.8 -24.4 69.6 -1.5 0 -2.9 AATATGTTAAGTTTTGAGTT
3865 SEQ ID NO: 903 -22.8 -24.4 69.6 -1.5 0 -4 AAATATGTTAAGTTTTGAGT
3866 SEQ ID NO: 904 -22.8 -24.4 69.6 -1.5 0 -5.1 ATTTGGCCACCTTGAATAGA
3909 SEQ ID NO: 905 -22.8 -33.8 83.3 -9.4 0 -11.2
GCATTTCTTTGACCCCTACA 4166 SEQ ID NO: 906 -22.8 -34.9 85.1 -12.1 0 -2.7
TGTTGTCATTCAACTGCTGG 4535 SEQ ID NO: 907 -22.8 -34.6 84 -9.5 -2.3 -7.6
GTCCCATTGAGAGTGAAACT 375 SEQ ID NO: 908 -22.7 -35.1 84 -11.7 -0.5 -4.7 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
AGATACATCAGAGTGAGTTT
605 SEQ ID NO: 909 -22.7 -32.1 79.6 -9.4 0 -3.6
GTTGGTGCCAGTCTGGCCCT
644 SEQ ID NO: 910 -22.7 -43.8 96.2 -12 -6.9 -26.4
TTGCCACCGTTGGTGCCAGT
652 SEQ ID NO: 911 -22.7 -41.5 94.4 -14.2 -4.3 -17
CCTTCCAAAAGGAATGAATC
829 SEQ ID NO: 912 -22.7 -31.4 79.1 -6 -2.7 -7.8
TTAGTGTCCGGTAAAATGAG
874 SEQ ID NO: 913 -22.7 -32 81.4 -9.3 0 -6.5
TGCTTAATTACCCCAGGGGT
994 SEQ ID NO: 914 -22.7 -38.3 91.1 -10.5 -2.8 -18.4
TTGCCTGACAGTAAACTGTG
1026 SEQ ID NO: 915 -22.7 -34.6 85.1 -8.6 -3.3 -12.1
TATAACACTTCAGGTTCAAT
1747 SEQ ID NO: 916 -22.7 -29.5 77.1 -6.8 0 -4.5
ATATAACACTTCAGGTTCAA
1748 SEQ ID NO: 917 -22.7 -29.5 77.1 -6.8 0 -4.5
TGAGCGTAGTCATGATCCTC
1803 SEQ ID NO: 918 -22.7 -37.5 86.1 -13.8 0 -10
GCATGTGTTTACATTGGTCG
2052 SEQ ID NO: 919 -22.7 -32.9 81.9 -9.5 -0.2 -8.7
TTGACAATGGCTTTTCCTAG
2221 SEQ ID NO: 920 -22.7 -32.4 81.7 -9.7 0 -3.8
TTAAGGAGATTTTCAACCAC
2314 SEQ ID NO: 921 -22.7 -30.3 78.4 -6.7 0 -9.8
TCTACTCTCCCATCACTGAA
2664 SEQ ID NO: 922 -22.7 -36.9 86.1 -14.2 0 -4.6
TCATCCAGCCAACTGTGAAA
2821 SEQ ID NO: 923 -22.7 -36 85.6 -11.1 -2.2 -7.2
TTCATCCAGCCAACTGTGAA
2822 SEQ ID NO: 924 -22.7 -36 85.6 -11.1 -2.2 -7.2
GTGCTCTATACCAGTTAGGA
3296 SEQ ID NO: 925 -22.7 -35.8 86.4 -12.2 -0.8 -4.6
TTTTGGGTAAAGTTTAGTGA
3565 SEQ ID NO: 926 -22.7 -28.8 77.7 -6.1 0 -3.8
CCTTCCCAGATTAGTGAATA
4027 SEQ ID NO: 927 -22.7 -33.2 82.2 -10.5 0 -3
GGTTATCCATCAGCATTTCT
4178 SEQ ID NO: 928 -22.7 -33.1 81.2 -9.4 -0.9 -5.2
CTCTTCCCCTAGAGCAAACT
4258 SEQ ID NO: 929 -22.7 -37.3 88.2 -12.2 -2.4 -7.2
ATATTGCTGGTACCTCTATG
4583 SEQ ID NO: 930 -22.7 -33.4 82.7 -9.4 0 -10.6
CATATTGCTGGTACCTCTAT
4584 SEQ ID NO: 931 -22.7 -33.4 82.7 -9.4 0 -10.6
TTATTTGGAAATAAACTCTT
4664 SEQ ID NO: 932 -22.7 -24.5 69.6 -1.8 0 -5.2
TTTATTTGGAAATAAACTCT
4665 SEQ ID NO: 933 -22.7 -24.5 69.6 -1.8 0 -6.4
CCATTGAGAGTGAAACTGCT
372 SEQ ID NO: 934 -22.6 -34.8 83.9 -12.2 0 -5.1 . .- . , .
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CTTTTGTTTCTGTCTCTCCC
402 SEQ ID NO: 935 -22.6 -33.6 82.7 -11 0 0
TTCGGAACCAACGGGAATTG
1199 SEQ ID NO: 936 -22.6 -35.3 85.3 -11.4 -1.2 -6.8
TGGAGGAGAGCTTACATCTG
1331 SEQ ID NO: 937 -22.6 -37.8 87.5 -13.9 -0.3 -10.6
GTGGTCCTGTTGTTGCTGTT
1362 SEQ ID NO: 938 -22.6 -36.7 87.6 -14.1 0 -5.6
TCCACAAGTTAAGACTCCAT
1430 SEQ ID NO: 939 -22.6 -34 83.3 -11.4 0 -4.9
CTGCTCTTTTGAAGAAAACT
1458 SEQ ID NO: 940 -22.6 -29.7 77.7 -6.4 0 -9
TTCATTCCAGCCTGAAGACA
1573 SEQ ID NO: 940 -22.6 -36.2 85.6 -12.9 -0.4 -6.8
CCTGGTATTGCCTTTGCCCA
1861 SEQ ID NO: 942 -22.6 -39.1 91.4 -13.7 -2.8 -10.7
TCCAGGTGTAAGTTCCTGAA
1882 SEQ ID NO: 943 -22.6 -36.2 86.8 -8.8 -4.8 -12.2
TGGTCATCCAGGTGTAAGTT
1888 SEQ ID NO: 944 -22.6 -36.2 86.6 -12 -1.6 -6.4
AGTCATTCTCTGCTCATTAA
2018 SEQ ID NO: 945 -22.6 -32 79.6 -9.4 0 -1.7
TTTACATTGGTCGTACATGC
2045 SEQ ID NO: 946 -22.6 -32.1 81.2 -9.5 0 -6.1
TATAAACCACATGTAGTGCG
2599 SEQ ID NO: 947 -22.6 -32.2 81.9 -8.5 -0.9 -9.1
ATCTACTCTCCCATCACTGA
2665 SEQ ID NO: 948 -22.6 -37.1 85.8 -14.5 0 -2.8
TAATTCTCCACTGAAGCAGA
3254 SEQ ID NO: 949 -22.6 -34.2 83.1 -10.9 -0.5 -7.2
TCACTGGACTAGGTGCTCTA
3308 SEQ ID NO: 950 -22.6 -39.1 90.3 -14.2 -2.3 -8.9
ACAGGGCCTCTTGGTAGTTA
3377 SEQ ID NO: 951 -22.6 -38.5 91 -13.5 -2.4 -10.9
GCAAAAATAGGGCGTTAGGT
3401 SEQ ID NO: 952 -22.6 -33.9 85.1 -10.8 -0.1 -6.5
CTTTGTCTGATTAAAAGTCT
3536 SEQ ID NO: 953 -22.6 -27.9 74.7 -5.3 0 -4.5
TTAACCAATTGGTGACAGAT
3636 SEQ ID NO: 954 -22.6 -31.5 79.9 -5.1 -3.1 -15.6
ATTAACCAATTGGTGACAGA
3637 SEQ ID NO: 955 -22.6 -31.5 79.9 -5.1 -3.1 -15.6
TTGTGTTAAACTCTATGGCA
3754 SEQ ID NO: 956 -22.6 -31.5 80.9 -8.4 -0.2 -4.3
ACAGGACTTGTGTTAAACTC
3761 SEQ ID NO: 957 -22.6 -32.5 82 -9 -0.5 -9.5
TGTTTGGTTTCTGAGACCAT
4239 SEQ ID NO: 958 -22.6 -33.6 82.6 -4.9 -4.2 -20.4
GAGCAAACTGTTTGGTTTCT
4247 SEQ ID NO: 959 -22.6 -32 81.3 -7.4 -1.8 -11.5
GTCTGTGGTATACAATTTCA
674 SEQ ID NO: 960 -22.5 -30.8 78.9 -8.3 0 -7.1 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target imolecular molecul Site oligo binding formation Duplex structure oligo oligo
CAGTTTTCATCTATCAACAG
781 SEQ ID NO: 961 -22.5 -29.3 76.2 -6.8 0 -1.5
GTCTTCTCCCGCCAGAGGAG
809 SEQ ID NO: 962 -22.5 -42.6 93.1 -15 -5.1 -15.5
GATGAAATCTTCTTTTTCTG
965 SEQ ID NO: 963 -22.5 -26.9 72 -3.6 -0.6 -4.1
CCAATTATATTTGCTCCAGG
1051 SEQ ID NO: 964 -22.5 -31.6 80.4 -9.1 0 -4.6
AGTCAAGTTGTCATCTCCAG
1241 SEQ ID NO: 965 -22.5 -35.1 83.8 -12.6 0 -3.7
TGGTCCTGTTGTTGCTGTTG
1361 SEQ ID NO: 966 -22.5 -36.6 87.5 -14.1 0 -5.6
TCAGATCAGGAGCAAAACAC
1995 SEQ ID NO: 967 -22.5 -35.1 83.9 -12.6 0 -5.2
CTCATTAATAATCAGATCAG
2006 SEQ ID NO: 968 -22.5 -27.8 73.1 -5.3 0 -5.2
TTTCATACAGAGATACTCTT
2114 SEQ ID NO: 969 -22.5 -30 77 -6.6 -0.8 -7.9
TTTTCATACAGAGATACTCT
2115 SEQ ID NO: 970 -22.5 -30 77 -6.6 -0.7 -7
ACGACTCAACTGCTTCTGTT
2636 SEQ ID NO: 971 -22.5 -35.1 84.8 -12.6 0 -2.5
GGTTTCTAATTTCTGGGATA
2706 SEQ ID NO: 972 -22.5 -30.8 79 -6.4 -1.9 -6.8
TCTTTGACTGTGGAGATTAC
2739 SEQ ID NO: 973 -22.5 -33.1 81.5 -8.9 -1.7 -7.1
ACTATACAGGGGGGGGATAC
2864 SEQ ID NO: 974 -22.5 -40.4 92.5 -17.9 0 -1.6
TTTCAGGTTTCCATGCATAA
2902 SEQ ID NO: 975 -22.5 -31.4 79.9 -8.9 0 -7
TTTTCAGGTTTCCATGCATA
2903 SEQ ID NO: 976 -22.5 -31.4 79.9 -8.9 0 -7
TGCAAAAATAGGGCGTTAGG
3402 SEQ ID NO: 977 -22.5 -33.8 85 -10.8 -0.1 -6.5
TCACAGGACTTGTGTTAAAC
3763 SEQ ID NO: 978 -22.5 -32.5 82 -6.7 -1.5 -14.7
AATTTGGCCACCTTATAG
3910 SEQ ID NO: 979 -22.5 -32.3 81.7 -8.3 0 -11
TATTGCTGGTACCTCTATGC
4582 SEQ ID NO: 980 -22.5 -35.7 86.2 -11.8 -0.7 -10.6
TTGTAGGATAGAAAGGAATT
4643 SEQ ID NO: 981 -22.5 -29.6 77.2 -7.1 0 -1.3
CACCGTCCGCAGTTCCCGCC
74 SEQ ID NO: 982 -22.4 -45 97.1 -22.1 -0.1 -3.1
CTCGCCCGCCACCGTCCGCA
83 SEQ ID NO: 983 -22.4 -47.7 100.2 -24.2 -1 -4.6
CTCCTCTTAGGGTTTTATAG
219 SEQ ID NO: 984 -22.4 -31.9 81.7 -8.3 -1.1 -7.4
AAACCTTCACAGTAGCTCCT
237 SEQ ID NO: 985 -22.4 -36 86.9 -13.6 0 -5.1
CAGAAACCTTCACAGTAGCT
240 SEQ ID NO: 986 -22.4 -34.8 85.1 -12.4 0 -3 , ..
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
ACAGTCCCATTGAGAGTGAA
378 SEQ ID NO: 987 -22.4 -36.3 85.5 -12.2 -1.6 -10.8
TATACAGTCCCATTGAGAGT
381 SEQ ID NO: 988 -22.4 -34.6 83.7 -12.2 0 -4.4
CCCGAGGAAAGGCTGATTTG
457 SEQ ID NO: 989 -22.4 -37.3 87.6 -14.3 -0.3 -6.4
TGAGGTTTGCAATGCTTTCT
501 SEQ ID NO: 990 -22.4 -33.3 82.6 -8.8 -1.2 -12.2
ATCTGTCCTCCAGAGGTACT
1105 SEQ ID NO: 991 -22.4 -39.1 89.3 -13.4 -3 -14.4
ACATCTGTCCTCCAGAGGTA
1107 SEQ ID NO: 992 -22.4 -39.1 89.2 -13.4 -3.3 -11.8
CCAACGGGAATTGGTGGAAT
1192 SEQ ID NO: 993 -22.4 -36.1 86.2 -12.1 -1.6 -9.3
GGGAGGTGGTCCTGTTGTTG
1367 SEQ ID NO: 994 -22.4 -40.4 91.8 -14.9 -3.1 -8.1
CCAGGCAGAGTTTGGGAGGT
1380 SEQ ID NO: 995 -22.4 -41.5 93.4 -14.5 -4.6 -14.5
TTCCACAAGTTAAGACTCCA
1431 SEQ ID NO: 996 -22.4 -33.8 83.6 -11.4 0 -4.9
CTATATGATCTCCACCCCAG
1954 SEQ ID NO: 997 -22.4 -37 86.3 -14.6 0 -4.2
AGCATGTGTTTACATTGGTC
2053 SEQ ID NO: 998 -22.4 -32.6 81.5 -9.5 -0.2 -8.6
GATTTTCAACCACTTCATGC
2307 SEQ ID NO: 999 -22.4 -31.4 79.1 -9 0 -2.2
TTCCATTTGAATATTTTGGT
2415 SEQ ID NO: 1000 -22.4 -26.7 72.5 -3.5 -0.6 -7.6
CAAAACAACAGATGAAAACA
2570 SEQ ID NO: 1001 -22.4 -27.3 73.8 -4.9 0 -2.5
CCACATGTAGTGCGTATTTA
2593 SEQ ID NO: 1002 -22.4 -32.2 81.9 -8.5 -0.9 -10.2
GCCATCCTTCTTTGACTGTG
2747 SEQ ID NO: 003 -22.4 -36 85.5 -13.6 0 -1.7
ATAGCTAAGTGCCATCAGGT
3155 SEQ ID NO: 1004 -22.4 -36.5 87.1 -12.5 -1.4 -10.3
TGACAAGAATACTGGAGATT
3199 SEQ ID NO: 1005 -22.4 -32 79.6 -9.6 0 -2.8
TTAAATTAGTCTTTTGCAAC
3353 SEQ ID NO: 1006 -22.4 -25.4 71.9 -3 0 -6.2
GCTTAGTAAATATGTTAAGT
3873 SEQ ID NO: 1007 -22.4 -26.2 73.5 -1.5 -2.2 -7.8
ACATACTTTAGTGCAAGGGG
4427 SEQ ID NO: 1008 -22.4 -34.8 86 -12.4 0.4 -7.9
GGGAGGGCTGTATGTGAAAG
4515 SEQ ID NO: 1009 -22.4 -38.5 89.3 -16.1 0 -4.5
CGCTCGCCCGCCACCGTCCG
85 SEQ ID NO: 1010 -22.3 -48 100.3 -25 -0.5 -5.3
AAAAGTCTTCGCTGCTTGGA
298 SEQ ID NO:1011 -22.3 -34.9 85 -12.1 -0.1 -8.1
ATACAGTCCCATTGAGAGTG
380 SEQ ID NO: 1012 -22.3 -35.4 84.4 -12.2 -0.5 -9.1
I0l kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
ATGAAATCTTCTTTTTCTGT
964 SEQ ID NO: 1013 -22.3 -26.7 71.8 -3.6 -0.6 -3.8
GTCAAGTTGTCATCTCCAGA
1240 SEQ ID NO: 1014 -22.3 -35.4 83.7 -13.1 0 -5.4
GGTGGTCCTGTTGTTGCTGT
1363 SEQ ID NO: 1015 -22.3 -39.1 90.6 -15.7 -1 -5.6
TCCTGAAGCTTCATCAGAGC
1403 SEQ ID NO: 1016 -22.3 -37.7 86.7 -11.8 -2.4 -15.3
TTTTTCTTTGTTTTTCGAGC
1600 SEQ ID NO: 1017 -22.3 -26.4 72.9 -4.1 0 -5.2
TTCAGAGGTTTCTTGTGAGA
1652 SEQ ID NO: 1018 -22.3 -33.9 82.6 -88 -2.8 -12.3
TCATGATCCTCCAAGTTGAG
1794 SEQ ID NO: 1019 -22.3 -35.4 83.5 -12.6 -0.2 -6.6
AAGGTTTTCATACAGAGATA
2119 SEQ ID NO: 1020 -22.3 -29.7 77.1 -6.8 -0.3 -3.9
GACCGTCCTTAGGAACTGAA
2151 SEQ ID NO: 1021 -22.3 -36.8 87.1 -12.9 -1.2 -10.8
TTCAGACCGTCCTTAGGAAC
2155 SEQ ID NO: 1022 -22.3 -36.8 87.1 -12.9 -1.2 -11
AGTTCTGGCTGGAGTTTCCT
2247 SEQ ID NO: 1023 -22.3 -38.2 89.2 -13.2 -1 -13.5
GGCACTGGTGGTTTAGGTGC
2765 SEQ ID NO: 1024 -22.3 -40.4 92.8 -13.9 -4.2 -13.2
ATATTAGAGATTTAGTTTTT
3581 SEQ ID NO: 1025 -22.3 -22.7 66.2 0.4 0 -6.3
CACAGGACTTGTGTTAAACT
3762 SEQ ID NO: 1026 -22.3 -32.2 82 -6.7 -1.4 -14.5
AAGTTGGTAAGGTGCACACA
3839 SEQ ID NO: 1027 -22.3 -35.6 87.1 -11.8 0 -11
AGAGCAAACTGTTTGGTTTC
4248 SEQ ID NO: 1028 -22.3 -32 81.3 -7.4 -1.1 -12.7
CTGCTGGGGGAGGGCTGTAT
4522 SEQ ID NO: 1029 -22.3 -44.2 96.6 -20.7 -1.1 -8.5
CGCAGTTCCCGCCGCAGCCG
67 SEQ ID NO: 1030 -22.2 -46.2 98.8 -22 -2 -7.5
TATAGAAGTCCATCACATCT
204 SEQ ID NO: 1031 -22.2 -32.6 80.1 -10.4 0 -2.6
AACCTTCACAGTAGCTCCTC
236 SEQ ID NO: 1032 -22.2 -37.5 88.1 -15.3 0 -6
TCTGTGGGGGCAGCAGACAC
571 SEQ ID NO: 1033 -22.2 -44.1 95.8 -17.9 -4 -12
TCTGAAGATACATCAGAGTG
610 SEQ ID NO: 1034 -22.2 -33.5 80.9 -7.6 -3.7 -10.4
TCCTGCAAAATGTCAAAGGT
700 SEQ ID NO: 1035 -22.2 -33.3 82.7 -10.2 -0.8 -5.8
TTCGACCAGGGAAGTTCAGA
1272 SEQ ID NO: 1036 -22.2 -37.8 87.9 -15.1 -0.1 -6
GAGAGCTTACATCTGGTCTC
1326 SEQ ID NO: 1037 -22.2 -37 85.9 -12.5 -2.3 -7.8
GTTAAGACTCCATAATGACA
1423 SEQ ID NO: 1038 -22.2 -31 78.8 -8.8 0 -3.5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CGATAGCGGCATGCTGGGCA
1549 SEQ ID NO: 1039 -22.2 -42.7 94.1 -15.6 -2.6 -17.9
TTCCAGCCTGAAGACATTTT
1569 SEQ ID NO: 1040 -22.2 -33.5 82.6 -11.3 0 -6.8
CACTGCTGCAATCACTTGCC
1838 SEQ ID NO: 1041 -22.2 -38 88.4 -13.9 -1.9 -7.9
TAAGTTCCTGAAACCTGGTA
1874 SEQ ID NO: 1042 -22.2 -33.9 84.6 -11 -0.4 -8
CTGCTCATTAATAATCAGAT
2009 SEQ ID NO: 1043 -22.2 -28.8 74.9 -6.6 0 -6.4
TAAGGTTTTCATACAGAGAT
2120 SEQ ID NO: 1044 -22.2 -29.7 77.1 -6.8 -0.5 -3.1
AAGCAGTAAGGTTTTCATAC
2126 SEQ ID NO: 1045 -22.2 -30.3 79.3 -8.1 0 -4.8
AGAGAGAAGCAGTAAGGTTT
2132 SEQ ID NO: 1046 -22.2 -33.8 83.6 -11.6 0 -5.4
TTTCCTTCCCTCTTGACAAT
2233 SEQ ID NO: 1047 -22.2 -33.7 82.6 -11.5 0 -2.1
GAGATTTTCAACCACTTCAT
2309 SEQ ID NO: 1048 -22.2 -30.4 77.3 -8.2 0 -6.7
CTCGGGGAATTCAATACTCA
2366 SEQ ID NO: 1049 -22.2 -34.6 83.4 -10.8 -1.5 -8.1
AGGACAAACTGATAGTTTAT
2522 SEQ ID NO: 1050 -22.2 -29.5 77.2 -6.4 -0.3 -9.4
TAGCTAAGTGCCATCAGGTT
3154 SEQ ID NO: 1051 -22.2 -36.3 87.4 -12.5 -1.5 -10.4
TCTTCTGATAGCTAAGTGCC
3162 SEQ ID NO: 1052 -22.2 -35.6 85.5 -12.5 -0.8 -7.5
CGTTAGGTACAGACAGGGCC
3389 SEQ ID NO: 1053 -22.2 -40.1 92.3 -16.5 -1.3 -7.3
TTAAAAGTCTCTCAGCTGTG
3526 SEQ ID NO: 1054 -22.2 -32.6 81.7 -9.7 0 -9.1
GACTTGTGTTAAACTCTATG
3757 SEQ ID NO: 1055 -22.2 -29.4 77.4 -6.7 -0.1 -2.8
CTTCCCTTCCCAGATTAGTG
4031 SEQ ID NO: 1056 -22.2 -36.2 86.3 -14 0 -3
ACTATACAAGCTATTTTACA
4088 SEQ ID NO: 1057 -22.2 -27.4 75.2 -5.2 0 -5
CACTATACAAGCTATTTTAC
4089 SEQ ID NO: 1058 -22.2 -27.4 75.4 -5.2 0 -5
ACACTATACAAGCTATTTTA
4090 SEQ ID NO: 1059 -22.2 -27.4 75.2 -5.2 0 -5
TACACTATACAAGCTATTTT
4091 SEQ ID NO: 1060 -22.2 -27.4 75.2 -5.2 0 -5
TTACACTATACAAGCTATTT
4092 SEQ ID NO: 1061 -22.2 -27.4 75.2 -5.2 0 -5
TTTACACTATACAAGCTATT
4093 SEQ ID NO: 1062 -22.2 -27.4 75.2 -5.2 0 -5
GTTTGGTTTCTGAGACCATC
4238 SEQ ID NO: 1063 -22.2 -33.9 82.5 -5.6 -4.2 -20.4
AGCAAACTGTTTGGTTTCTG
4246 SEQ ID NO: 1064 -22.2 -31.7 81.3 -7.4 -2.1 -10.6 kcal/mol kcal/mol deg C kcal/mol kcal/mol - kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecul Site oligo binding formation Duplex structure oligo oligo
TGGTACCTCTATGCAAACTA
4576 SEQ ID NO: 1065 -22.2 -34.3 85 -10.8 0 -10.6
ATAATTTCTCCAAAATACTG
4740 SEQ ID NO: 1066 -22.2 -25.9 71.5 -3.7 0 0
CGCAGAAACCTTCACAGTAG
242 SEQ ID NO: 1067 -22.1 -35.1 85.4 -13 0 -2.8
GACGCAGAAACCTTCACAGT
244 SEQ ID NO: 1068 -22.1 -36.3 86.3 -14.2 0 -2.8
GATTGAGAAGCGACAGCCAG
274 SEQ ID NO: 1069 -22.1 -37.8 87.2 -14.6 -1 -4.6
TCATTTCCCATCACTTTTGT
415 SEQ ID NO: 1070 -22.1 -31.3 79 -9.2 0 0
GCCAGTCTGGCCCTTCAAAT
638 SEQ ID NO: 1071 -22.1 -39.3 90.4 -11.3 -3.7 -20
GGGTTTAGTGTCCGGTAAAA
878 SEQ ID NO: 1072 -22.1 -34 85.3 -8.7 -3.2 -8
TGAAATCTTCTTTTTCTGTT
963 SEQ ID NO: 1073 -22.1 -26.5 71.9 -3.6 -0.6 -3.8
AACTGTGCCCAGTTTCTCTT
1013 SEQ ID NO: 1074 -22.1 -35.6 86.2 -11.1 -2.3 -12.1
CCTGACAGTAAACTGTGCCC
1023 SEQ ID NO: 1075 -22.1 -38.2 89.7 -12.8 -3.3 -12.1
TCCAGATCCTTGGCACCTAT
1226 SEQ ID NO: 1076 -22.1 -38.9 89.2 -15.3 -1.4 -7
AAGTCAAGTTGTCATCTCCA
1242 SEQ ID NO: 1077 -22.1 -33.9 82.4 -11.8 0 -3.7
GTTACCAGGATTTTCAGAGG
1664 SEQ ID NO: 1078 -22.1 -33.9 83.3 -11.8 0 -4.3
TTTTCAACCACTTCATGCAT
2305 SEQ ID NO: 1079 -22.1 -31.1 79 -9 0 -5.3
ATTTTCAACCACTTCATGCA
2306 SEQ ID NO: 1080 -22.1 -31.1 79 -9 0 -3.1
CTTTGGGCACTGGTGGTTTA
2770 SEQ ID NO: 1081 -22.1 -36.6 88.7 -12.5 0 -12.2
GTTAGGTACAGACAGGGCCT
3388 SEQ ID NO: 1082 -22.1 -39.8 92.1 -15.9 -1.8 -9.7
TGATTAAAAGTCTCTCAGCT
3529 SEQ ID NO: 1083 -22.1 -31.8 79.8 -9.7 0 -3.7
TAAAGTTTAGTGAGAGGAAT
3558 SEQ ID NO: 1084 -22.1 -29.5 77.3 -7.4 0 -3.2
GTGACAGATGGGAATGTGAA
3625 SEQ ID NO: 1085 -22.1 -35.3 83.6 -12.1 -1 -4.3
CGCATTGCTTACTGAGCCTT
332 SEQ ID NO: 1086 -22 -36.2 87 -12.4 -1.4 -11.4
GGCCCTGCTGTGGGAATCCC
438 SEQ ID NO: 1087 -22 -45.3 96.9 -18.8 -4.5 -13.8
CAAAGGTGCTTTGGTCTGTG
687 SEQ ID NO: 1088 -22 -35.3 85.9 -11.7 -1.6 -9.6
AAATGTCAAAGGTGCTTTGG
693 SEQ ID NO: 1089 -22 -31.8 81.1 -8.1 -1.7 -9.6
TTCCAAAAGGAATGAATCGT
827 SEQ ID NO: 1090 -22 -30.6 77.8 -6 -2.6 -10.1 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecu Site oligo binding formation Duplex structure oligo oligo
CTGTTTTCACTTGGGGCAGT
948 SEQ ID NO: 1091 -22 -36.5 87.7 -11.3 -3.2 -11.9
TTTCGGAACCAACGGGAATT
1200 SEQ ID NO: 1092 -22 -34.1 83.9 -9.4 -2.7 -7.6
CAGGCAGAGTTTGGGAGGTG
1379 SEQ ID NO: 1093 -22 -40.3 91.7 -16.9 -1.3 -7.9
TCACTGCTGCAATCACTTGC
1839 SEQ ID NO: 1094 -22 -37.1 86.9 -13.9 -1.1 -7.9
GTTCCTGAAACCTGGTATTG
1871 SEQ ID NO: 1095 -22 -33.7 83.4 -11 -0.4 -8
AAGTTCCTGAAACCTGGTAT
1873 SEQ ID NO: 1096 -22 -33.7 83.6 -11 -0.3 -8
TTGGTCATCCAGGTGTAAGT
1889 SEQ ID NO: 1097 -22 -36.2 86.6 -12 -2.2 -7.6
GCCATAAGAAACATCCAGGA
1927 SEQ ID NO: 1098 -22 -35.3 84.6 -13.3 0 -5.8
CCCAGAGCAAATGCCATAAG
1939 SEQ ID NO: 1099 -22 -35.9 86.1 -13.1 -0.6 -6.1
TCTCGGGGAATTCAATACTC
2367 SEQ ID NO: 1100 -22 -34.9 83.4 -11.9 0 -9.9
GAAAAAAAGTATGAAGAGAA
2805 SEQ ID NO: 1101 -22 -25.7 71 -3.7 0 -1
TTTTTTTCAGGTTTCCATGC
2906 SEQ ID NO: 1102 -22 -29.6 77.7 -7.6 0 -5.3
GTAAATATGTTAAGTTTTGA
3868 SEQ ID NO: 1103 -22 -23.6 68.7 -1.5 0 -5.1
CTTAGGGTTTTATAGAAGTC
214 SEQ ID NO: 1104 -21.9 -29.6 78.4 -6.9 0 -9.4
GATCTGGCTGCTGCGCATTG
345 SEQ ID NO: 1105 -21.9 -39.9 89.8 -16.4 -1.6 -10.3
TCTGGCCCTTCAAATGTTGC
633 SEQ ID NO: 1106 -21.9 -36.9 87.4 -14.5 0 -7.5
CCTGCAAAATGTCAAAGGTG
699 SEQ ID NO: 1107 -21.9 -33 82.6 -10.2 -0.7 -5.6
CTTGCTTAATTACCCCAGGG
996 SEQ ID NO: 1108 -21.9 -35.8 87.6 -12.7 -1.1 -9
TGCCTGACAGTAAACTGTGC
1025 SEQ ID NO: 1109 -21.9 -37.1 88.2 -11.9 -3.3 -12.1
CCAGGAAAGCTTGCCTGACA
1036 SEQ ID NO: 1110 -21.9 -39.1 90.5 -10.8 -4.6 -21
GTGGTACATCTGTCCTCCAG
1112 SEQ ID NO: 1111 -21.9 -38.9 89 -15.4 -1.5 -8.5
AGTGGTACATCTGTCCTCCA
1113 SEQ ID NO: 1112 -21.9 -38.9 89.3 -15.4 -1.5 -8.5
GAAGTCAAGTTGTCATCTCC
1243 SEQ ID NO: 1113 -21.9 -34.2 82.3 -12.3 0 -5.5
AGAAGTCAAGTTGTCATCTC
1244 SEQ ID NO: 1114 -21.9 -33 80.6 -11.1 0 -6.5
ACCAGGCAGAGTTTGGGAGG
1381 SEQ ID NO: 1115 -21.9 -41.5 93.4 -14.5 -5.1 -15.3
CCTGAAGCTTCATCAGAGCA
1402 SEQ ID NO: 1116 -21.9 -37.4 86.8 -11.8 -3.3 -15.3 ,
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target moleculair molecul Site oligo binding formation Duplex structure oligo oligo
ATGATGCAATCATTCCTTCC
1507 SEQ ID NO: 1117 -21.9 -33.1 80.1 -10.6 -0.1 -8.5
TTTCGATAGCGGCATGCTGG
1552 SEQ ID NO: 1118 -21.9 -38.1 88.5 -11.3 -2.6 -17.9
TGTTTTTCGAGCTTCCAGGT
1592 SEQ ID NO:1119 -21.9 -34.9 85.1 -13 0 -6.6
AGTTCCTGAAACCTGGTATT
1872 SEQ ID NO: 1120 -21.9 -33.7 83.6 -11 -0.6 -8
TCTATATGATCTCCACCCCA
1955 SEQ ID NO: 1121 -21.9 -37.3 86.5 -15.4 0 -4.2
CCCTCTTGACAATGGCTTTT
2226 SEQ ID NO: 1122 -21.9 -34.4 84.3 -12.5 0 -4.1
CTGCCAGTTCTGGCTGGAGT
2252 SEQ ID NO: 1123 -21.9 -41.6 93.1 -13.2 -6.3 -20.6
AGAATCCAAGAGTTTTGTCA
2285 SEQ ID NO: 1124 -21.9 -31.4 79.1 -9 0 -8
AAACAGCTGTACAATAACTT
3051 SEQ ID NO: 1125 -21.9 -28.9 77.5 -4.3 0 -13.6
CTTCTGATAGCTAAGTGCCA
3161 SEQ ID NO: 1126 -21.9 -35.3 85.5 -12.5 -0.8 -7.5
TGTTAATTTGCACAACCTAT
3275 SEQ ID NO: 1127 -21.9 -29 77.3 -7.1 0 -4.6
TGAGCTTTCCTGTACCATCA
3663 SEQ ID NO: 1128 -21.9 -36.4 86.4 -14.5 0 -6.4
TAAATATGTTAAGTTTTGAG
3867 SEQ ID NO: 1129 -21.9 -23.5 68.5 -1.5 0 -5.1
CTGAGACCATCGCTGCCTGT
4229 SEQ ID NO: 1130 -21.9 -41.2 91.6 -19.3 0 -4.5
AGTCCATCAGTGAATATCAA
120 SEQ ID NO: 1131 -21.8 -32.2 79.4 -10.4 0 -6.7
GTAGCTCCTCCTCTTAGGGT
226 SEQ ID NO: 1132 -21.8 -40.2 91.9 -16.6 -1.8 -7.6
AGATCTGGCTGCTGCGCATT
346 SEQ ID NO: 1133 -21.8 -39.9 90.3 -16.5 -1.5 -10.3
TTTCCCATCACTTTTGTTTC
412 SEQ ID NO: 1134 -21.8 -29.9 77.6 -8.1 0 0
AGTGGATGCTGAACTCTTGG
548 SEQ ID NO: 1135 -21.8 -37.2 87 -15.4 0 -4.6
AGCAGACACAGCAGTGGATG
560 SEQ ID NO: 1136 -21.8 -39.5 89.7 -14.1 -3.6 -12.5
CGTTGGTGCCAGTCTGGCCC
645 SEQ ID NO: 1137 -21.8 -44.1 96.3 -14.4 -5.7 -24
TATACAATTTCACATTGCCA
666 SEQ ID NO: 1138 -21.8 -29.4 76.9 -7.6 0 -2.2
TCTGTGGTATACAATTTCAC
673 SEQ ID NO: 1139 -21.8 -30.8 78.9 -8.3 -0.5 -7.1
GCAAAATGTCAAAGGTGCTT
696 SEQ ID NO: 1140 -21.8 -31.9 81.1 -9.4 -0.3 -8.2
TCGTCTTCTCCCGCCAGAGG
811 SEQ ID NO: 1141 -21.8 -42.9 93.7 -19.1 -2 -9.1
CTTCCAAAAGGAATGAATCG
828 SEQ ID NO: 1142 -21.8 -30.5 77.8 -6 -2.7 -9.2 . ., . ,
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
ATTTTGGGTTTAGTGTCCGG
883 SEQ ID NO: 1143 -21.8 -33.7 84.2 -8.5 -3.4 -6.8
GATGCTGTATTCATGTCATA
1132 SEQ ID NO: 1144 -21.8 -31.4 78.3 -8.7 -0.8 -6.5
GTTCGACCAGGGAAGTTCAG
1273 SEQ ID NO: 1145 -21.8 -37.6 87.7 -15.3 -0.1 -6
TGTTCGACCAGGGAAGTTCA
1274 SEQ ID NO: 1146 -21.8 -37.6 88 -15.3 -0.1 -6.5
AAGAAAACTTTACAGCTTCC
1447 SEQ ID NO: 1147 -21.8 -29.9 78.7 -8.1 0 -4
CTCTTGACAATGGCTTTTCC
2224 SEQ ID NO: 1148 -21.8 -33.5 82.5 -11.7 0 -3.8
ACAAAACAACAGATGAAAAC
2571 SEQ ID NO: 1149 -21.8 -27.4 74 -5.6 0 -2.5
ATTAAGGTTTCTAATTTCTG
2711 SEQ ID NO: 1150 -21.8 -25.7 71.7 -3.9 0 -4
CTTCTTTGACTGTGGAGATT
2741 SEQ ID NO: 1151 -21.8 -32.6 80.7 -8.9 -1.9 -8
GACAGTAATAGCTATAAAAG
2961 SEQ ID NO: 1152 -21.8 -27.6 75.3 -4.9 0 -9.8
TTAAAACCAGACAGTAATAG
2970 SEQ ID NO: 1153 -21.8 -28.2 76.4 -6.4 0 -1.1
TATACCAGTTAGGACTGTTA
3290 SEQ ID NO: 1154 -21.8 -32.3 82.5 -8.3 -1.9 -12.2
GGTGCTCTATACCAGTTAGG
3297 SEQ ID NO: 1155 -21.8 -36.7 87.9 -12.2 -2.7 -8.4
TAATTTGGCCACCTTGAATA
3911 SEQ ID NO: 1156 -21.8 -31.5 80.8 -8.3 0 -10.8
GAAATAAACTCTTGTTGTAG
4657 SEQ ID NO: 1157 -21.8 -26.9 73.9 -5.1 0 -2.8
ATGATTCTTTGGAGTCCATC
132 SEQ ID NO: 1158 -21.7 -33.2 80.3 -7.7 -3.8 -14.2
GCATTGCTTACTGAGCCTTT
331 SEQ ID NO: 1159 -21.7 -34.7 85 -11.2 -1.4 -11.4
CATTTCCCATCACTTTTGTT
414 SEQ ID NO: 1160 -21.7 -29.8 77.4 -8.1 0 0
TCCCCCGAGGAAAGGCTGAT
460 SEQ ID NO: 1161 -21.7 -42.4 93.7 -19.2 -1.4 -7.3
AAGTCTGTTTCCCCCGAGGA
469 SEQ ID NO: 1162 -21.7 -39.9 91.2 -17.3 -0.8 -7.2
GGCCCTTCAAATGTTGCTGT
630 SEQ ID NO: 1163 -21.7 -36.7 87.5 -14.5 0 -7.5
GAATGAATCGTCTTCTCCCG
818 SEQ ID NO: 1164 -21.7 -35.1 82.9 -13.4 0 -5.2
GCTTAATTACCCCAGGGGTG
993 SEQ ID NO: 1165 -21.7 -38.3 90.6 -10.5 -3.8 -20.4
TGACAGTAAACTGTGCCCAG
1021 SEQ ID NO: 1166 -21.7 -37 88.2 -12 -3.3 -12.1
TATTACCAATTATATTTGCT
1056 SEQ ID NO: 1167 -21.7 -25.2 71 -3.5 0 -4.6
CCTGCTGTTGAGAAAGGGAT
1149 SEQ ID NO: 1168 -21.7 -37.1 87.2 -13.8 -1.5 -6.7 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecul Site oligo binding formation Duplex structure oligo oligo
ACCAACGGGAATTGGTGGAA
1193 SEQ ID NO: 1169 -21.7 -37.2 88.2 -12.1 -3.4 -9.8
ATTCCAGCCTGAAGACATTT
1570 SEQ ID NO: 1170 -21.7 -33.7 82.4 -12 0 -6.8
ATAACACTTCAGGTTCAATA
1746 SEQ ID NO: 1171 -21.7 -29.5 77.1 -7.8 0 -4.2
TTCCTTCCCTCTTGACAATG
2232 SEQ ID NO: 1172 -21.7 -34.9 84 -13.2 0 -2.1
CAGTTCTGGCTGGAGTTTCC
2248 SEQ ID NO: 1173 -21.7 -38.2 88.8 -13.2 -3 -14.1
CAGTTGATAAAACCGCTGCC
2267 SEQ ID NO: 1174 -217 -35 85.5 -12.4 -0.7 -5.4
ATTTCAGCTAACATCTCGGG
2380 SEQ ID NO: 1175 -21.7 -34.4 83.5 -127 0 -57
AAAAAAGTATGAAGAGAAAG
2803 SEQ ID NO: 1176 -21.7 -25.4 70.9 -3.7 0 -1
TTTTTTCAGGTTTCCATGCA
2905 SEQ ID NO: 1177 -21.7 -30.8 79.3 -9.1 0 -5.3
CTGTACAATAACTTGAATAA
3045 SEQ ID NO: 1178 -217 -26 72.4 -4.3 0 -7.2
AAAGTTTAGTGAGAGGAATT
3557 SEQ ID NO: 1179 -21.7 -29.1 76.6 -7.4 0 -3.4
TTTGGGTAAAGTTTAGTGAG
3564 SEQ ID NO: 1180 -21.7 -30 79.5 -8.3 0 -3.8
GATTAACCAATTGGTGACAG
3638 SEQ ID NO: 1181 -217 -31.5 79.8 -6.2 -2.4 -15.3
TATGGCACACATTAGGGATG
3741 SEQ ID NO: 1182 -21.7 -35.4 85.1 -13.7 0 -47
AGTAAATATGTTAAGTTTTG
3869 SEQ ID NO: 1183 -21.7 -23.3 68.5 -1.5 0 -5.1
TCCCAGATTAGTGAATACCA
4024 SEQ ID NO: 1184 -21.7 -34.5 83.8 -12.8 0 -3
CAGCATTTCTTTGACCCCTA
4168 SEQ ID NO: 1185 -217 -34.8 85 -13.1 0 -2.7
GGGTTTTATAGAAGTCCATC
210 SEQ ID NO: 1186 -21.6 -32.1 80.7 -7.3 -3.1 -13.4
ACGCAGAAACCTTCACAGTA
243 SEQ ID NO: 1187 -21.6 -35.2 85.7 -13.6 0 -2.8
GAGGGTGAAGACGCAGAAAC
253 SEQ ID NO: 1188 -21.6 -37.7 87.7 -14.9 -1.1 -4.5
TTTCTGTCTCTCCCATATAC
396 SEQ ID NO: 1189 -21.6 -33.6 82.1 -12 0 -0.7
TATTGAGGTTTGCAATGCTT
504 SEQ ID NO: 1190 -21.6 -31.2 79.9 -8.8 -0.5 -8.7
CAGACACAGCAGTGGATGCT
558 SEQ ID NO: 1191 -21.6 -39.5 897 -14.1 -3.8 -12.9
TGTTCTGAAGATACATCAGA
613 SEQ ID NO: 1192 -21.6 -32.3 79.4 -7.6 -3.1 -9.2
TGGCCCTTCAAATGTTGCTG
631 SEQ ID NO: 1193 -21.6 -36.6 87.4 -14.5 0 -7.5
CTGGCCCTTCAAATGTTGCT
632 SEQ ID NO: 1194 -21.6 -36.6 87.4 -14.5 0 -7.5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CACATTGCCACCGTTGGTGC
656 SEQ ID NO: 1195 -21.6 -39.3 90.6 -14.6 -2.9 -13.6
TTTTTCTGTTTTCACTTGGG
953 SEQ ID NO: 1196 -21.6 -29.4 77.8 -7.8 0 -4
TCGATGAAATCTTCTTTTTC
967 SEQ ID NO: 1197 -21.6 -27.5 72.5 -5.9 0 -4.6
TACCCCAGGGGTGCAGAGTT
986 SEQ ID NO: 1198 -21.6 -42.9 96 -14.9 -4.1 -21
GGGATGCTGTATTCATGTCA
1134 SEQ ID NO: 1199 -21.6 -35.6 84.2 -11.4 -1.4 -13.4
AACGGGAATTGGTGGAATGA
1190 SEQ ID NO: 1200 -21.6 -35.2 84.6 -13.6 0 -3.8
AATTTTCGGAACCAACGGGA
1203 SEQ ID NO: 1201 -21.6 -34.1 83.9 -10.4 -2.1 -7.2
TCCTGTTGTTGCTGTTGAGG
1358 SEQ ID NO: 1202 -21.6 -36.8 87.4 -15.2 0 -4.1
GTCCTGTTGTTGCTGTTGAG
1359 SEQ ID NO: 1203 -21.6 -35.7 85.7 -14.1 0 -1.7
AGTTAAGACTCCATAATGAC
1424 SEQ ID NO: 1204 -21.6 -31 78.9 -9.4 0 -3.5
CCTTCCACTGCTCTTTTGAA
1465 SEQ ID NO: 1205 -21.6 -34.5 84.4 -12.9 0 -1.7
CAGCCTGAAGACATTTTCGA
1566 SEQ ID NO: 1206 -21.6 -34.1 82.8 -11.8 -0.3 -8.1
ATGTTGAGCGTAGTCATGAT
1807 SEQ ID NO: 1207 -21.6 -33.6 81.5 -11.4 0 -8.5
ATTGCCTTTGCCCATTTCAC
1855 SEQ ID NO: 1208 -21.6 -34.4 84.2 -12.8 0 -2
TCTCTGCTCATTAATAATCA
2012 SEQ ID NO: 1209 -21.6 -30.1 76.7 -8.5 0 -2.5
GTCATTCTCTGCTCATTAAT
2017 SEQ ID NO: 1210 -21.6 -31 77.8 -9.4 0 -1.7
ACATCTCGGGGAATTCAATA
2370 SEQ ID NO: 1211 -21.6 -33.6 81.7 -10.6 0 -10.7
TTTGAATATTTTGGTATCTG
2410 SEQ ID NO: 1212 -21.6 -25.9 71.4 -4.3 0 -5.4
GTGATGACGACTCAACTGCT
2642 SEQ ID NO: 1213 -21.6 -36.8 85.7 -15.2 0 -5.6
TGTGGAGATTACGTCCACAT
2731 SEQ ID NO: 1214 -21.6 -35.8 85 -77 -6.5 -16
GGTGGTTTAGGTGCCATCCT
2759 SEQ ID NO: 1215 -21.6 -39.5 91.2 -10.9 -7 -16.4
TGGTGGTTTAGGTGCCATCC
2760 SEQ ID NO:1216 -21.6 -39.5 91.2 -10.9 -7 -16.4
AATTTCATCCAGCCAACTGT
2825 SEQ ID NO: 1217 -21.6 -33.5 82.4 -11.1 -0.6 -3.9
TTGTAAACTTTTTTTCAGGT
2914 SEQ ID NO: 1218 -21.6 -26 73.1 -4.4 0 -1.2
GATTAAAAGTCTCTCAGCTG
3528 SEQ ID NO: 1219 -21.6 -31.8 79.8 -9.7 0 -7.9
AAGAAATTCACAGGACTTGT
3770 SEQ ID NO: 1220 -21.6 -31.2 79.3 -8.9 0 -8.7 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TACAGAAAGTTGGTAAGGTG
3845 SEQ ID NO: 1221 -21.6 -32.5 82.8 -10.9 0 -3.2
TATGTTAAGTTTTGAGTTTA
3863 SEQ ID NO: 1222 -21.6 -24.6 70.5 -3 0 -3.1
CTTAGTAAATATGTTAAGTT
3872 SEQ ID NO: 1223 -21.6 -23.7 69.4 -1.5 -0.3 -5.1
CAAACTGTTTGGTTTCTGAG
4244 SEQ ID NO: 1224 -21.6 -30.7 79.4 -7.7 -1.1 -10.3
TTACATACTTTACATACTTT
4438 SEQ ID NO: 1225 -21.6 -25 71.3 -3.4 0 0
ACTCTTGTTGTAGGATAGAA
4650 SEQ ID NO: 1226 -21.6 -32 80.8 -9.8 -0.1 -8.2
AGCCGAGATAAACAACTTAG
52 SEQ ID NO: 1227 -21.5 -32 81.4 -10.5 0 -1.7
TGGCTGCTGCGCATTGCTTA
341 SEQ ID NO: 1228 -21.5 -39.6 91.6 -14.2 -1.8 -16
TAAGTCTGTTTCCCCCGAGG
470 SEQ ID NO: 1229 -21.5 -38.8 90.4 -17.3 0 -4.9
AGTCTGGCCCTTCAAATGTT
635 SEQ ID NO: 1230 -21.5 -35.7 86 -13.7 0 -7.5
TGCCAGTCTGGCCCTTCAAA
639 SEQ ID NO: 1231 -21.5 -40.3 92.2 -12.9 -37 -20
TTTAGTGTCCGGTAAAATGA
875 SEQ ID NO: 1232 -21.5 -30.8 79.8 -9.3 0 -7.3
CAGACATTTTATTACCAATT
1065 SEQ ID NO: 1233 -21.5 -257 71.5 -4.2 0 0
GTCCTCCAGAGGTACTCACA
1101 SEQ ID NO: 1234 -21.5 -40.2 90.8 -15.4 -3 -14.4
TGTCCTCCAGAGGTACTCAC
1102 SEQ ID NO: 1235 -21.5 -40.2 90.8 -15.4 -3 -14.4
CCACAAGTTAAGACTCCATA
1429 SEQ ID NO: 1236 -21.5 -32.9 82.4 -11.4 0 -4.6
TTTTCGAGCTTCCAGGTTCA
1589 SEQ ID NO: 1237 -21.5 -35.1 84.9 -9.3 -4.3 -12.7
GTTTTCATACAGAGATACTC
2116 SEQ ID NO: 1238 -21.5 -30.1 77.2 -77 -07 -3.6
GAGAAGCAGTAAGGTTTTCA
2129 SEQ ID NO: 1239 -21.5 -32.6 81.9 -9.7 -1.3 -8.9
GTAGGTCATTCTAATTTCAT
2192 SEQ ID NO: 1240 -21.5 -287 75.3 -6.2 -0.9 -4.7
ATGTAGGTCATTCTAATTTC
2194 SEQ ID NO: 1241 -21.5 -28.7 75.3 -6.2 -0.9 -47
GGCTGGAGTTTCCTTCCCTC
2241 SEQ ID NO: 1242 -21.5 -40.8 91.7 -15.6 -3.6 -14.4
AGATTACGTCCACATATTAA
2726 SEQ ID NO: 1243 -21.5 -29.2 76.5 -7.7 0 -2.8
TAAACTTTTTTTCAGGTTTC
2911 SEQ ID NO: 1244 -21.5 -25 71.3 -2.2 -1.2 -5.5
GCGTTAGGTACAGACAGGGC
3390 SEQ ID NO: 1245 -21.5 -40.2 92.3 -18 -0.5 -7.3
GAGCTTTCCTGTACCATCAG
3662 SEQ ID NO: 1246 -21.5 -36.4 86.1 -14.1 -0.6 -6.5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
ATGTTAAGTTTTGAGTTTAC
3862 SEQ ID NO: 1247 -21.5 -25.5 72 -4 0 -3.1
GTCATTCAACTGCTGGGGGA
4531 SEQ ID NO: 1248 -21.5 -39.6 90.1 -17.4 0 -9
TTTTATTTGGAAATAAACTC
4666 SEQ ID NO: 1249 -21.5 -23.3 67.8 -1.8 0 -7.3
CTGGCAGAGGAGCCGCTCGC
98 SEQ ID NO: 1250 -21.4 -46.1 97.4 -20.2 -4.5 -14.6
TAGGGTTTTATAGAAGTCCA
212 SEQ ID NO: 1251 -21.4 -32 81.8 -7.3 -3.3 -13.4
GAGGTTTGCAATGCTTTCTT
500 SEQ ID NO: 1252 -21.4 -32.1 81.1 -8.8 -1 -11.8
AACCAACGGGAATTGGTGGA
1194 SEQ ID NO: 1253 -21.4 -37.2 88.2 -12.1 -3.7 -10.4
GAACCAACGGGAATTGGTGG
1195 SEQ ID NO: 1254 -21.4 -37.2 87.9 -12.1 -3.7 -10.4
GGAACCAACGGGAATTGGTG
1196 SEQ ID NO: 1255 -21.4 -37.2 87.9 -12.1 -3.7 -10.4
TGCTCTTTTGAAGAAAACTT
1457 SEQ ID NO: 1256 -21.4 -28.5 75.9 -6.4 0 -9
TGTCCTTCCACTGCTCTTTT
1468 SEQ ID NO: 1257 -21.4 -35.8 86.1 -14.4 0 -1.7
TTCGAGCTTCCAGGTTCATT
1587 SEQ ID NO: 1258 -21.4 -35.3 84.6 -9,3 -4.6 -13
TTTCGAGCTTCCAGGTTCAT
1588 SEQ ID NO: 1259 -21.4 -35.3 84.6 -9.3 -4.6 -13
TTTTTTCTTTGTTTTTCGAG
1601 SEQ ID NO: 1260 -21.4 -23.9 68.8 -2.5 0 -3.9
TTACCAGGATTTTCAGAGGT
1663 SEQ ID NO: 1261 -21.4 -33.9 83.5 -11.8 -0.5 -6.7
GAGCGTAGTCATGATCCTCC
1802 SEQ ID NO: 1262 -21.4 -387 87.5 -15.7 -0.3 -11.4
ACCTGGTATTGCCTTTGCCC
1862 SEQ ID NO: 1263 -21.4 -39.2 91.5 -15 -2.8 -10.7
TGCAGTAGGGTCATTTGGTC
1903 SEQ ID NO: 1264 -21.4 -37.4 88 -15.4 -0.3 -7.7
GAAGCAGTAAGGTTTTCATA
2127 SEQ ID NO: 1265 -21.4 -30.5 79.2 -8.1 -0.9 -7.4
TCCATTTGAATATTTTGGTA
2414 SEQ ID NO: 1266 -21.4 -27.1 73.3 -4.9 -0.6 -7.6
CCAGACAGTAATAGCTATAA
2964 SEQ ID NO: 1267 -21.4 -31.2 80.4 -8.9 0 -9.8
AGTCAATTTTTGTGGTCTTC
3177 SEQ ID NO: 1268 -21.4 -30 77.6 -8.6 0 -2.4
TTTGAGTCAATTTTTGTGGT
3181 SEQ ID NO: 1269 -21.4 -28.5 75.7 -6.6 0 -8.2
TTGCACAACCTATATAATTC
3268 SEQ ID NO: 1270 -21.4 -28.6 76.1 -7.2 0 -4.4
GAGGAATTACTTTGTCTGAT
3545 SEQ ID NO: 1271 -21.4 -30.8 78.1 -8.4 -0.9 -6.2
ATTCACAGGACTTGTGTTAA
3765 SEQ ID NO: 1272 -21.4 -31.4 80 -6.7 -1.5 -14.7 .
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
AATTCACAGGACTTGTGTTA
3766 SEQ ID NO: 1273 -21.4 -31.4 80 -6.7 -1.5 -14.7
AAAGTTGGTAAGGTGCACAC
3840 SEQ ID NO: 1274 -21.4 -34.4 85.5 -11.5 0 -11
TGGCTTAGTAAATATGTTAA
3875 SEQ ID NO: 1275 -21.4 -27.3 75.2 -5.9 0 -5.1
TTCCCAGATTAGTGAATACC
4025 SEQ ID NO: 1276 -21.4 -33.3 82.3 -11.9 0 -3
ACATATTGCTGGTACCTCTA
4585 SEQ ID NO: 1277 -21.4 -34.5 84.7 -11.8 0 -10.6
AGGATAGAAAGGAATTAGTG
4639 SEQ ID NO: 1278 -21.4 -30.8 78.9 -9.4 0 -2.8
GCAGAGGAGCCGCTCGCCCG
95 SEQ ID NO: 1279 -21.3 -47.6 99.2 -22.8 -3.3 -14.6
ATATACAGTCCCATTGAGAG
382 SEQ ID NO: 1280 -21.3 -33.5 81.8 -12.2 0 -4.4
CATATACAGTCCCATTGAGA
383 SEQ ID NO: 1281 -21.3 -33.5 817 -12.2 0 -3.4
TCCTTCCAAAAGGAATGAAT
830 SEQ ID NO: 1282 -21.3 -31.4 79.1 -6 -4.1 -10.2
TTACATTACTGGGGCTTGAC
927 SEQ ID NO: 1283 -21.3 -35.1 85.9 -13.8 0 -5.9
GGGGCAGTGTTACATTACTG
936 SEQ ID NO: 1284 -21.3 -36.1 87.3 -11.5 -3.3 -10.1
TTTTCGGAACCAACGGGAAT
1201 SEQ ID NO: 1285 -21.3 -34.1 83.9 -9.4 -3.4 -9
ACTGCTCTTTTGAAGAAAAC
1459 SEQ ID NO: 1286 -21.3 -29.8 77.8 -7.8 0 -8.9
TTTTCTTTGTTTTTCGAGCT
1599 SEQ ID NO: 1287 -21.3 -27.6 74.6 -6.3 0 -5.2
GGCCTGCTGAATTCCTTTTA
1622 SEQ ID NO: 1288 -21.3 -34.8 85.1 -13 0 -8.3
TACCAGGATTTTCAGAGGTT
1662 SEQ ID NO: 1289 -21.3 -33.9 83.5 -11.8 -0.6 -7.6
CTCTGCTCATTAATAATCAG
2011 SEQ ID NO: 1290 -21.3 -29.8 76.8 -8.5 0 -4.4
AAGAGAGAAGCAGTAAGGTT
2133 SEQ ID NO: 1291 -21.3 -33.8 83.6 -12.5 0 -3.4
GAATTCAATACTCATGGTCT
2360 SEQ ID NO: 1292 -21.3 -31 77.8 -9.7 0 -7
AACATCTCGGGGAATTCAAT
2371 SEQ ID NO: 1293 -21.3 -33.2 81 -10.5 0 -10.7
GGACAAACTGATAGTTTATA
2521 SEQ ID NO: 1294 -21.3 -28.7 76.3 -6.4 -0.6 -9.7
GTGGAGATTACGTCCACATA
2730 SEQ ID NO: 1295 -21.3 -35 84.2 -7.7 -6 -15
CTTTTTTTCAGGTTTCCATG
2907 SEQ ID NO: 1296 -21.3 -28.3 75.8 -7 0 -5.3
TCCACTGAAGCAGATATATA
3248 SEQ ID NO: 1297 -21.3 -32.7 80.9 -10.9 -0.1 -7.2
ACCAATTGGTGACAGATGGG
3633 SEQ ID NO: 1298 -21.3 -37.1 87 -7.8 -8 -16 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CAGATTAGTGAATACCAATA
4021 SEQ ID NO: 1299 -21.3 -28.8 75.9 -7.5 0 -3
GGAGGGCTGTATGTGAAAGT
4514 SEQ ID NO: 1300 -21.3 -37.4 88 -16.1 0 -4.9
TTCTGAAGCTTCTGTTGTCA
4547 SEQ ID NO: 1301 -21.3 -337 82.6 -10.2 -07 -12.5
ATTGCTGGTACCTCTATGCA
4581 SEQ ID NO: 1302 -21.3 -36.5 87 -12.7 -2.5 -10.6
AGGGTTTTATAGAAGTCCAT
211 SEQ ID NO: 1303 -21.2 -31.8 80.8 -7.3 -3.3 -13.4
AGCGACAGCCAGTGAGGGTG
266 SEQ ID NO: 1304 -21.2 -43.2 94.8 -19.7 -2.3 -8.6
TTGAGGTTTGCAATGCTTTC
502 SEQ ID NO: 1305 -21.2 -32.1 81 -8.8 -1.2 -12.2
GCAGCAGACACAGCAGTGGA
562 SEQ ID NO: 1306 -21.2 -41.8 92.8 -18.6 -2 -9.1
GGTATACAATTTCACATTGC
668 SEQ ID NO: 1307 -21.2 -29.5 77.2 -8.3 0 -7.1
ATCTATCAACAGGTCTGATC
773 SEQ ID NO: 1308 -21.2 -33.7 80.9 -10.9 0 -11.1
AAACTGTGCCCAGTTTCTCT
1014 SEQ ID NO: 1309 -21.2 -35.6 86.2 -11.1 -3.2 -13.9
AAATGGCAGACATTTTATTA
1071 SEQ ID NO: 1310 -21.2 -26.9 73.3 -4.2 -1.4 -4.8
TCTGTCCTCCAGAGGTACTC
1104 SEQ ID NO: 1311 -21.2 -40.4 90.7 -15.9 -3 -14.4
ATGCTGTATTCATGTCATAG
1131 SEQ ID NO: 1312 -21.2 -31.1 78.2 -9.9 0 -6
TCCTGCTGTTGAGAAAGGGA
1150 SEQ ID NO: 1313 -21.2 -38.4 89 -15.2 -2 -9.3
TCAAGTTGTCATCTCCAGAT
1239 SEQ ID NO: 1314 -21.2 -34.3 82 -13.1 0 -3.9
TCGACCAGGGAAGTTCAGAG
1271 SEQ ID NO: 1315 -21.2 -39 89.1 -17.3 -0.1 -6
TTTCAGAGGTTTCTTGTGAG
1653 SEQ ID NO: 1316 -21.2 -32.4 81 -8.8 -2.4 -11.3
ACGTTGCAGGAACTATTGTT
1686 SEQ ID NO: 1317 -21.2 -32.7 82.4 -10.9 -0.3 -6
GAGTTGTGGTAACGTTGCAG
1697 SEQ ID NO: 1318 -21.2 -35 85.6 -10.6 -1.3 -14.6
AGGTGTAAGTTCCTGAAACC
1879 SEQ ID NO: 1319 -21.2 -34.8 85.4 -12 -1.5 -7.1
CCAGAGCAAATGCCATAAGA
1938 SEQ ID NO: 1320 -21.2 -35 84.6 -13.1 -0.4 -5.9
CATCCAGCCAACTGTGAAAA
2820 SEQ ID NO: 1321 -21.2 -34.5 84.1 -11.1 -2.2 -7.2
TTTATTTGGGAAAGCTACGA
3087 SEQ ID NO: 1322 -21.2 -30.7 79.8 -9.5 0 -5
ATACCAGTTAGGACTGTTAA
3289 SEQ ID NO: 1323 -21.2 -31.9 81.8 -8.3 -2.2 -12.5
ATTAAAAGTCTCTCAGCTGT
3527 SEQ ID NO: 1324 -21.2 -31.6 80 -9.7 0 -9.1 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target imolecular molecul Site oligo binding formation Duplex structure oligo oligo
TTCACAGGACTTGTGTTAAA
3764 SEQ ID NO: 1325 -21.2 -31.2 80.3 -67 -1.5 -14.7
GTTGTCATTCAACTGCTGGG
4534 SEQ ID NO: 1326 -21.2 -35.8 85.5 -12.3 -2.3 -7.6
GGTACCTCTATGCAAACTAT
4575 SEQ ID NO: 1327 -21.2 -33.3 83.1 -10.8 0 -10.6
CTCTTGTTGTAGGATAGAAA
4649 SEQ ID NO: 1328 -21.2 -30.7 79 -8.9 -0.1 -8.2
TTCTCCAAAATACTGAAAAT
4735 SEQ ID NO: 1329 -21.2 -26.8 72.6 -5.6 0 -1.1
CCCCCGAGGAAAGGCTGATT
459 SEQ ID NO: 1330 -21.1 -40.9 92.3 -19.2 -0.3 -7.2
CTTTGGTCTGTGGTATACAA
679 SEQ ID NO: 1331 -21.1 -32.7 82.5 -11.6 0 -7.1
GGTTTAGTGTCCGGTAAAAT
877 SEQ ID NO: 1332 -21.1 -31.8 81.7 -9.7 -0.9 -8
TTACCCCAGGGGTGCAGAGT
987 SEQ ID NO: 1333 -21.1 -42.9 96 -14.9 -4.3 -21.9
TAGTGGTACATCTGTCCTCC
1114 SEQ ID NO: 1334 -21.1 -38.1 88.5 -15.4 -1.5 -8.5
ATGGAGGAGAGCTTACATCT
1332 SEQ ID NO: 1335 -21.1 -36.8 86 -14.7 -0.1 -9.9
AACGTTGCAGGAACTATTGT
1687 SEQ ID NO: 1336 -21.1 -32.7 82.4 -10.9 -0.4 -67
TTGCCTTTGCCCATTTCACT
1854 SEQ ID NO: 1337 -21.1 -35.4 86.1 -14.3 0 -2
TTCTGGCTGGAGTTTCCTTC
2245 SEQ ID NO: 1338 -21.1 -37.2 87.4 -13.2 -1 -13.9
GTTCTGGCTGGAGTTTCCTT
2246 SEQ ID NO: 1339 -21.1 -37 87.6 -13.2 -1 -13.5
TAGAATCCAAGAGTTTTGTC
2286 SEQ ID NO: 1340 -21.1 -30.6 78.3 -9 -0.1 -8
GGGGAATTCAATACTCATGG
2363 SEQ ID NO: 1341 -21.1 -34.2 82.8 -11.8 -1.2 -7
GTGGTTTAGGTGCCATCCTT
2758 SEQ ID NO: 1342 -21.1 -37.1 88.3 -11.7 -4.3 -11.8
TAGAAAATTTCATCCAGCCA
2830 SEQ ID NO: 1343 -21.1 -31.6 79.8 -10.5 0 -6
CTAGAAAATTTCATCCAGCC
2831 SEQ ID NO: 1344 -21.1 -31.6 79.8 -10.5 0 -6
TCTGATAGCTAAGTGCCATC
3159 SEQ ID NO: 1345 -21.1 -35.8 85.1 -12.5 -2.2 -7.2
GGAGATTTGAGTCAATTTTT
3186 SEQ ID NO: 1346 -21.1 -27.9 73.8 -5.2 -0.7 -11
CTTTTGCAACCATCATCCAC
3343 SEQ ID NO: 1347 -21.1 -33.5 82.3 -12.4 0 -6.2
TTTAAATTAGTCTTTTGCAA
3354 SEQ ID NO: 1348 -21.1 -24.1 69.6 -3 0 -4.7
TTTTAAATTAGTCTTTTGCA
3355 SEQ ID NO: 1349 -21.1 -24.1 69.6 -3 0 -2.9
TGGTAGTTATTTTTTAAATT
3366 SEQ ID NO: 1350 -21.1 -22 66.2 -0.8 0 -2.7 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GCCATTTTTGCCATATTAGA
3593 SEQ ID NO: 1351 -21.1 -30.5 78.9 -9.4 0 -1.4
TTGGTAAGGTGCACACAGAA
3836 SEQ ID NO: 1352 -21.1 -35.8 86.9 -13.2 0 -11
TAGAGCAAACTGTTTGGTTT
4249 SEQ ID NO: 1353 -21.1 -30.9 80.4 -7.4 -1.1 -12.9
TCTGTTGTCATTCAACTGCT
4537 SEQ ID NO: 1354 -21.1 -33.7 82.4 -10.3 -2.3 -7.6
CCGAGATAAACAACTTAGCT
50 SEQ ID NO: 1355 -21 -32 81.4 -11 0 -3.1
TTATAGAAGTCCATCACATC
205 SEQ ID NO: 1356 -21 -31.4 78.5 -10.4 0 -2.2
ACCAAAAGTCTTCGCTGCTT
301 SEQ ID NO: 1357 -21 -347 85.1 -13.7 0 -5.2
AACCAAAAGTCTTCGCTGCT
302 SEQ ID NO: 1358 -21 -34.7 85.1 -13.7 0 -5.2
CAACCAAAAGTCTTCGCTGC
303 SEQ ID NO: 1359 -21 -34.7 84.8 -13.7 0 -3.2
GATGCTGAACTCTTGGGGTT
544 SEQ ID NO: 1360 -21 -37.2 87.2 -14 -2.1 -11.6
TAAAATGAGAGGCTTGCAGT
863 SEQ ID NO: 1361 -21 -33.7 83.4 -12.2 0 -8.1
TCTTTTTCTGTTTTCACTTG
955 SEQ ID NO: 1362 -21 -27.3 74 -6.3 0 0
CTTCTTTTTCTGTTTTCACT
957 SEQ ID NO: 1363 -21 -27.3 74.1 -6.3 0 0
TCGGAACCAACGGGAATTGG
1198 SEQ ID NO: 1364 -21 -37.7 88.4 -15.1 -1.5 -5.4
GGAGAGCTTACATCTGGTCT
1327 SEQ ID NO: 1365 -21 -37.9 87.6 -16.3 -0.3 -7.6
TAAGACTCCATAATGACATC
1421 SEQ ID NO: 1366 -21 -31.4 78.5 -10.4 0 -3.5
GAGTCATTCTCTGCTCATTA
2019 SEQ ID NO: 1367 -21 -33.5 81.1 -9.4 -3.1 -8.9
TTTCAACCACTTCATGCATA
2304 SEQ ID NO: 1368 -21 -31.5 79.7 -10.5 0 -7
TATTAAGGTTTCTAATTTCT
2712 SEQ ID NO: 1369 -21 -24.9 70.5 -3.9 0 -37
TTCTTTGACTGTGGAGATTA
2740 SEQ ID NO: 1370 -21 -31.8 79.9 -8.9 -1.9 -8
TGTAAACTTTTTTTCAGGTT
2913 SEQ ID NO: 1371 -21 -26 73.1 -4.4 -0.3 -3
TTTTTTTGACAAGAATACTG
3205 SEQ ID NO: 1372 -21 -25.2 70.9 -3.7 0 -7.8
CCTATATAATTCTCCACTGA
3260 SEQ ID NO: 1373 -21 -31.4 79.3 -10.4 0 -3.4
AAGTTTAGTGAGAGGAATTA
3556 SEQ ID NO: 1374 -21 -29.5 77.3 -8.5 0 -3.6
CTGAGCTTTCCTGTACCATC
3664 SEQ ID NO: 1375 -21 -36.4 86.1 -15.4 0 -6.6
TAGCTGAGCTTTCCTGTACC
3667 SEQ ID NO: 1376 -21 -37.6 89 -14.8 0 -11.8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TGTTAAGTTTTGAGTTTACA
3861 SEQ ID NO: 1377 -21 -26.5 73.8 -5.5 0 -5
ATACAAGCTATTTTACAATC
4085 SEQ ID NO: 1378 -21 -26.2 72.5 -5.2 0 -47
AATAATTTCTCCAAAATACT
4741 SEQ ID NO: 1379 -21 -24.7 69.5 -37 0 0
GCTGTGGGAATCCCAGGTCA
432 SEQ ID NO: 1380 -20.9 -42 92.9 -15.9 -4.1 -18.4
GAAATCTTCTTTTTCTGTTT
962 SEQ ID NO: 1381 -20.9 -25.3 70.2 -3.6 -0.6 -3.8
GAGAAAGGGATGCTGTATTC
1140 SEQ ID NO: 1382 -20.9 -34.4 82.9 -12.3 0 -10.3
TCCAATTTTCGGAACCAACG
1206 SEQ ID NO: 1383 -20.9 -32.9 82.1 -10.4 -1.6 -6.6
GGAGGTGGTCCTGTTGTTGC
1366 SEQ ID NO: 1384 -20.9 -40.5 91.7 -177 -1.9 -7.6
CTGAAGCTTCATCAGAGCAC
1401 SEQ ID NO: 1385 -20.9 -36.3 85.1 -11.7 -3.1 -15.3
GAAGAAAACTTTACAGCTTC
1448 SEQ ID NO: 1386 -20.9 -29 76.9 -8.1 0 -5.5
TTCCACTGCTCTTTTGAAGA
1463 SEQ ID NO: 1387 -20.9 -33.6 82.7 -127 0 -6.8
CTGTCCTTCCACTGCTCTTT
1469 SEQ ID NO: 1388 -20.9 -37 87.4 -16.1 0 -17
CCAGCACATAGGTAATTGTG
1489 SEQ ID NO: 1389 -20.9 -33.8 83.8 -10.8 -2.1 -6.1
GGTGTAAGTTCCTGAAACCT
1878 SEQ ID NO: 1390 -20.9 -34.8 85.4 -12 -1.9 -9.1
AGAGATACTCTTCATAAGAT
2106 SEQ ID NO: 1391 -20.9 -30.4 76.7 -8.8 -0.5 -6.8
TCCTAGCTCTTTGATGTAGG
2207 SEQ ID NO: 1392 -20.9 -35.5 85.5 -13 -1.6 -8.9
ACAACAGATGAAAACAATAA
2566 SEQ ID NO: 1393 -20.9 -267 72.6 -5.8 0 -2.5
TTCTGATAGCTAAGTGCCAT
3160 SEQ ID NO: 1394 -20.9 -34.3 83.8 -12.5 -0.8 -7.5
AAACTCTATGGCACACATTA
3747 SEQ ID NO: 1395 -20.9 -317 80.6 -10.8 0 -4.1
TAAACTCTATGGCACACATT
3748 SEQ ID NO: 1396 -20.9 -31.7 80.6 -10.8 0 -4.2
TTAAACTCTATGGCACACAT
3749 SEQ ID NO: 1397 -20.9 -31.7 80.6 -10.8 0 -4.2
GGACTTGTGTTAAACTCTAT
3758 SEQ ID NO: 1398 -20.9 -30.6 79.2 -9 -0.5 -3.7
TAGTAAATATGTTAAGTTTT
3870 SEQ ID NO: 1399 -20.9 -22.5 67.1 -1.5 0 -4.8
TTAGTAAATATGTTAAGTTT
3871 SEQ ID NO: 1400 -20.9 -22.5 67.1 -1.5 0 -5.1
CTATACAAGCTATTTTACAA
4087 SEQ ID NO: 1401 -20.9 -26.1 73.3 -5.2 0 -5
TTTACATACTTTAGTGCAAG
4430 SEQ ID NO: 1402 -20.9 -28 76.4 -7.1 0 -6.3 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GTTTACATACTTTACATACT
4440 SEQ ID NO: 1403 -20.9 -26.3 73.5 -5.4 0 -2.6
GTTTTATAGAAGTCCATCAC
208 SEQ ID NO: 1404 -20.8 -29.8 77.3 -9 0 -6.5
AGAAACCTTCACAGTAGCTC
239 SEQ ID NO: 1405 -20.8 -35.1 85.1 -14.3 0 -3.9
GGCTGCTGCGCATTGCTTAC
340 SEQ ID NO: 1406 -20.8 -39.7 91.2 -15.5 -1.6 -14.9
ACAGATCTGGCTGCTGCGCA
348 SEQ ID NO: 1407 -20.8 -42.2 93.2 -17.3 -1.5 -16.4
AACACTGGTCGACCTATTGA
518 SEQ ID NO: 1408 -20.8 -35.6 85.4 -10.5 -1 -16.7
ACTCTTGGGGTTCTCTGGAA
536 SEQ ID NO: 1409 -20.8 -38.4 89.1 -14.5 -3.1 -127
ATGCTGAACTCTTGGGGTTC
543 SEQ ID NO: 1410 -20.8 -37.2 87.2 -12.1 -3.7 -16.6
GTTCTGAAGATACATCAGAG
612 SEQ ID NO: 1411 -20.8 -32.3 79.5 -7.6 -3.9 -10.6
AGGGATGCTGTATTCATGTC
1135 SEQ ID NO: 1412 -20.8 -35.6 84.2 -12.2 -0.4 -13.4
GCACCTATTCCAATTTTCGG
1214 SEQ ID NO: 1413 -20.8 -32.9 82.1 -12.1 0 -27
GGCACCTATTCCAATTTTCG
1215 SEQ ID NO: 1414 -20.8 -32.9 82.1 -12.1 0 -3.9
TTTTCAGAGGTTTCTTGTGA
1654 SEQ ID NO: 1415 -20.8 -31.2 79.4 -8.8 -1.6 -9.9
GTTGAGTCTGGAACAGAGCT
1780 SEQ ID NO: 1416 -20.8 -37.4 87.2 -15 -1.4 -10.6
AGAGTCATTCTCTGCTCATT
2020 SEQ ID NO: 1417 -20.8 -34.3 82 -10.2 -3.3 -8.9
GATTTCAGCTAACATCTCGG
2381 SEQ ID NO: 1418 -20.8 -33.5 81.7 -12.7 0 -6.1
TTGATATTTCCATTTGAATA
2422 SEQ ID NO: 1419 -20.8 -25.1 69.2 -4.3 0 -4.1
CTATAAACCACATGTAGTGC
2600 SEQ ID NO: 1420 -20.8 -31.9 81.4 -10 -0.9 -9.4
ATCCTTCTTTGACTGTGGAG
2744 SEQ ID NO: 1421 -20.8 -35 83.9 -12.6 -1.6 -7
GCTATCCTAACTATACAGGG
2873 SEQ ID NO: 1422 -20.8 -34.8 85.4 -12.5 -1.4 -4.9
TTCAGGTTTCCATGCATAAA
2901 SEQ ID NO: 1423 -20.8 -31.4 79.9 -10.6 0 -7
AGTAATAGCTATAAAAGGCA
2958 SEQ ID NO: 1424 -20.8 -29.7 78.9 -6.4 -2.5 -9.8
AATACTGGAGATTTGAGTCA
3192 SEQ ID NO: 1425 -20.8 -32 79.6 -9.6 -0.7 -11
GACAGGGCCTCTTGGTAGTT
3378 SEQ ID NO: 1426 -20.8 -39.6 91.3 -16 -2.8 -10.9
TTGGGTAAAGTTTAGTGAGA
3563 SEQ ID NO: 1427 -20.8 -31.5 81.2 -10.7 0 -3.6
TTGCCATATTAGAGATTTAG
3586 SEQ ID NO: 1428 -20.8 -28.7 75.9 -7.9 0 -2.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TCTTGTGGCTTAGTAAATAT
3880 SEQ ID NO: 1429 -20.8 -29.6 78 -8.8 0 -5
TTTACATACTTTACATACTT
4439 SEQ ID NO: 1430 -20.8 -25 71.3 -4.2 0 0
GCCTTTAAGGATGTAAGCAC
4476 SEQ ID NO: 1431 -20.8 -33.9 84.3 -12.1 -0.7 -9.3
GTGCCTTTAAGGATGTAAGC
4478 SEQ ID NO: 1432 -20.8 -33.9 84.3 -12.1 -07 -9.3
TTTTATAGAAGTCCATCACA
207 SEQ ID NO: 1433 -20.7 -297 77 -9 0 -5.8
TTCTGTCTCTCCCATATACA
395 SEQ ID NO: 1434 -20.7 -34.8 83.6 -14.1 0 -0.7
TTCCCCCGAGGAAAGGCTGA
461 SEQ ID NO: 1435 -20.7 -42.2 94.1 -19.2 -2.3 -9.6
CTTGGGGTTCTCTGGAACAC
533 SEQ ID NO: 1436 -20.7 -38.2 88.9 -14.1 -3.4 -13
AGCAGTGGATGCTGAACTCT
551 SEQ ID NO: 1437 -20.7 -38.5 88.5 -14.5 -3.3 -11
ATCCTGCAAAATGTCAAAGG
701 SEQ ID NO: 1438 -20.7 -32.2 80.7 -11 -0.2 -4.9
AGACATTTTATTACCAATTA
1064 SEQ ID NO: 1439 -20.7 -24.9 70.3 -4.2 0 -0.1
GGGTGAGTTGTGGTAACGTT
1701 SEQ ID NO: 1440 -207 -36.2 87.5 -15.5 0 -5.4
TTGAGCGTAGTCATGATCCT
1804 SEQ ID NO: 1441 -20.7 -36 84.9 -14.5 0 -9.4
AAACCTGGTATTGCCTTTGC
1864 SEQ ID NO: 1442 -20.7 -34.4 85.4 -12.1 -1.1 -10.7
CATGTGTTTACATTGGTCGT
2051 SEQ ID NO: 1443 -20.7 -31.7 80.3 -10.4 0 -8.6
TCCTTCCCTCTTGACAATGG
2231 SEQ ID NO: 1444 -20.7 -37.3 87.1 -16.6 0 -3.3
TGCCAGTTCTGGCTGGAGTT
2251 SEQ ID NO: 1445 -20.7 -40.4 92.1 -13.2 -6.3 -20.6
CCAAGAGTTTTGTCAGTTGA
2280 SEQ ID NO: 1446 -20.7 -32.2 81 -11 0 -8
TCCAAGAGTTTTGTCAGTTG
2281 SEQ ID NO: 1447 -20.7 -32.2 81 -11 0 -8
TGGTATCTGATTGGTGATGA
2399 SEQ ID NO: 1448 -20.7 -34.6 82.6 -12.4 -1.4 -5.4
AGTGATGACGACTCAACTGC
2643 SEQ ID NO: 1449 -207 -36.8 85.7 -16.1 0 -5.5
GACTTTGGGCACTGGTGGTT
2772 SEQ ID NO: 1450 -20.7 -39 90.8 -16.6 0 -11.6
AGAAAATTTCATCCAGCCAA
2829 SEQ ID NO: 1451 -20.7 -31.2 79.1 -10.5 0 -6
TATATAATTCTCCACTGAAG
3258 SEQ ID NO: 1452 -20.7 -29 75.9 -7.6 -0.5 -4.2
ATAATTTGGCCACCTTGAAT
3912 SEQ ID NO: 1453 -207 -31.3 79.9 -9 0 -11.1
TAGTGAATACCAATATGATA
4016 SEQ ID NO: 1454 -20.7 -28.2 74.7 -7.5 0 -1.5 . ,
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
CAGACTCACTGTTGGAATGA 4380 SEQ ID NO: 1455 -20.7 -35.1 83.7 -13.7 -0.4 -5.3
TGTCCTCATTTTATTTGGAA 4674 SEQ ID NO: 1456 -207 -29.2 76.3 -6.9 -1.6 -6.6
AGTAGCTCCTCCTCTTAGGG 227 SEQ ID NO: 1457 -20.6 -40.1 91.9 -18 -1.4 -9.4
CGACCTATTGAGGTTTGCAA 509 SEQ ID NO: 1458 -20.6 -34 83.8 -9.6 -3.8 -12.7
TTGGGGTTCTCTGGAACACT 532 SEQ ID NO: 1459 -20.6 -38.2 89.2 -14.2 -3.4 -12.1
TGCTGAACTCTTGGGGTTCT 542 SEQ ID NO: 1460 -20.6 -38.2 89.1 -12.1 -4.9 -19
GATACATCAGAGTGAGTTTT 604 SEQ ID NO: 1461 -20.6 -30.9 78 -10.3 0 -3.5
TTCTGAAGATACATCAGAGT 611 SEQ ID NO: 1462 -20.6 -32.3 79.5 -7.6 -4.1 -11.2
TCTATCAACAGGTCTGATCT 772 SEQ ID NO: 1463 -20.6 -34.7 82.7 -12.5 0 -11.1
CATCTATCAACAGGTCTGAT 774 SEQ ID NO: 1464 -20.6 -33.4 80.9 -11.2 0 -11.1
GGTACATCTGTCCTCCAGAG 1110 SEQ ID NO: 1465 -20.6 -39.1 88.9 -15.9 -2.6 -9.6
CAGATCAGGAGCAAAACACA 1994 SEQ ID NO: 1466 -20.6 -34.8 83.9 -14.2 0 -5.2
TTTGTCAGTTGATAAAACCG 2272 SEQ ID NO: 1467 -20.6 -29.2 77.1 -5.3 -3.1 -14
GGGAATTCAATACTCATGGT 2362 SEQ ID NO: 1468 -20.6 -33.1 81.3 -12.5 0 -6.7
GAATATTTTGGTATCTGATT 2407 SEQ ID NO: 1469 -20.6 -26.4 71.5 -5.8 0 -5.4 ATTTGAATATTTTGGTATCT
2411 SEQ ID NO: 1470 -20.6 -24.9 69.3 -4.3 0 -5.4 CATTTGAATATTTTGGTATC
2412 SEQ ID NO: 1471 -20.6 -24.9 697 -4.3 0 -6.1 TGCTATCCTAACTATACAGG
2874 SEQ ID NO: 1472 -20.6 -33.6 83.7 -12.5 -0.2 -2.5
AGCTAAGTGCCATCAGGTTA 3153 SEQ ID NO: 1473 -20.6 -36.3 87.4 -14 -1.7 -8.7
AAGTCTCTCAGCTGTGTTAC 3522 SEQ ID NO: 1474 -20.6 -35.2 84.9 -13.9 0 -9.1
GCTATTTTACAATCATTTTA 4079 SEQ ID NO: 1475 -20.6 -23.7 68.5 -3.1 0 -17
AGCATTTCTTTGACCCCTAC 4167 SEQ ID NO: 1476 -20.6 -34.9 85.1 -14.3 0 -2.7
CTCCATCTCCCTCTTCCCCT 4268 SEQ ID NO: 1477 -20.6 -42.3 92.7 -217 0 0
TCCATCAGTGAATATCAACT 118 SEQ ID NO: 1478 -20.5 -32.2 79.4 -11.7 0 -6.7
GTCCATCACATCTCCCCTCT 197 SEQ ID NO: 1479 -20.5 -40.3 89.5 -19.8 0 -0.4
TCCTCCTCTTAGGGTTTTAT 221 SEQ ID NO: 1480 -20.5 -34.2 84.4 -12.1 -1.6 -7.3 .
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecul Site oligo binding formation Duplex structure oligo oligo
AGACGCAGAAACCTTCACAG
245 SEQ ID NO: 1481 -20.5 -36.2 86.3 -15.7 0 -2.8
TGAGAAGCGACAGCCAGTGA
271 SEQ ID NO: 1482 -20.5 -40.1 90.5 -18.5 -1 -7.1
TGCTGCGCATTGCTTACTGA
337 SEQ ID NO: 1483 -20.5 -37.5 88.4 -15.1 -1 -11.9
CAGATCTGGCTGCTGCGCAT
347 SEQ ID NO: 1484 -20.5 -41.1 91.3 -16.5 -1.6 -16.4
ATTTCCCATCACTTTTGTTT
413 SEQ ID NO: 1485 -20.5 -28.6 75.6 -8.1 0 0
TTCCTTCCAAAAGGAATGAA
831 SEQ ID NO: 1486 -20.5 -31.2 79.3 -6 -4.7 -11.6
GTAAAATGAGAGGCTTGCAG
864 SEQ ID NO: 1487 -20.5 -33.7 83.2 -12.6 -0.1 -8.4
TTATTACCAATTATATTTGC
1057 SEQ ID NO: 1488 -20.5 -24 69.3 -3.5 0 -4.6
GAAATGGCAGACATTTTATT
1072 SEQ ID NO: 1489 -20.5 -28 74.5 -4.2 -3.3 -8.6
ACAGAAATGGCAGACATTTT
1075 SEQ ID NO: 1490 -20.5 -31.1 79.1 -77 -2.9 -7.8
CCTCCAGAGGTACTCACACC
1099 SEQ ID NO: 1491 -20.5 -41.1 92.1 -18.7 -1.6 -11.6
ACTTTACAGCTTCCACAAGT
1441 SEQ ID NO: 1492 -20.5 -33.4 83.8 -12.9 0 -4.6
CTTTGCCCATTTCACTGCTG
1850 SEQ ID NO: 1493 -20.5 -35.4 85.6 -14.9 0 -1.8
CAGGGTAGAGTCATTCTCTG
2026 SEQ ID NO: 1494 -20.5 -36.6 85.9 -13 -3 -13.5
GTTTACATTGGTCGTACATG
2046 SEQ ID NO: 1495 -20.5 -30.9 79.5 -10.4 0 -6.1
AGAGAAGCAGTAAGGTTTTC
2130 SEQ ID NO: 1496 -20.5 -32.6 82 -10.8 -1.2 -8.8
GCTCTTCAGACCGTCCTTAG
2159 SEQ ID NO: 1497 -20.5 -38 88.3 -17.5 0 -67
TCGGGGAATTCAATACTCAT
2365 SEQ ID NO: 1498 -20.5 -33.6 81.7 -11.5 -1.5 -7.1
CATCTCGGGGAATTCAATAC
2369 SEQ ID NO: 1499 -20.5 -33.6 81.6 -117 0 -10.7
TTCAGCTAACATCTCGGGGA
2378 SEQ ID NO: 1500 -20.5 -38.1 88.4 -16.8 0 -9.4
TGATATTTCCATTTGAATAT
2421 SEQ ID NO: 1501 -20.5 -25.3 69.1 -4.3 -0.2 -4.1
TCTTGGCCCTCTATAAACCA
2610 SEQ ID NO: 1502 -20.5 -36.4 87.5 -15.4 0 -7.5
ACTCAACTGCTTCTGTTGCC
2633 SEQ ID NO: 1503 -20.5 -36.9 87.4 -14.6 -1.8 -5.8
CATCTACTCTCCCATCACTG
2666 SEQ ID NO: 1504 -20.5 -36.8 85.6 -16.3 0 0
AAAAAAAGTATGAAGAGAAA
2804 SEQ ID NO: 1505 -20.5 -24.2 68.8 -3.7 0 -1
GTGAAAAAAAGTATGAAGAG
2807 SEQ ID NO: 1506 -20.5 -26.7 73 -6.2 0 -1 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TTTGGGAAAGCTACGAACTA
3083 SEQ ID NO: 1507 -20.5 -33 83.5 -12.5 0 -5
GAGTTTATTTGGGAAAGCTA
3090 SEQ ID NO: 1508 -20.5 -30.4 79.3 -8.3 -1.6 -10.3
CACACATTAGGGATGTGTAG
3736 SEQ ID NO: 1509 -20.5 -34.1 83.6 -8.2 -5.3 -18
CTTGTGGCTTAGTAAATATG
3879 SEQ ID NO:1510 -20.5 -29.3 78 -8.8 0 -5
AGTGAATACCAATATGATAT
4015 SEQ ID NO: 1511 -20.5 -28 73.8 -7.5 0 -2.9
CTTCCCAGATTAGTGAATAC
4026 SEQ ID NO: 1512 -20.5 -32.1 80.5 -11.6 0 -3
AGCTATTTTACAATCATTTT
4080 SEQ ID NO: 1513 -20.5 -24.5 69.4 -4 0 -3.9
CTGTAATCTCACTGGGTCAG
4322 SEQ ID NO: 1514 -20.5 -36.4 86 -14.9 -0.9 -87
TACATATTGCTGGTACCTCT
4586 SEQ ID NO: 1515 -20.5 -34.5 84.7 -12.7 0 -10.6
ATGTCCTCATTTTATTTGGA
4675 SEQ ID NO: 1516 -20.5 -29.4 76.1 -8 -0.8 -47
GCCGAGATAAACAACTTAGC
51 SEQ ID NO: 1517 -20.4 -33.3 83 -11 -1.9 -57
GTCTGATTGAGAAGCGACAG
278 SEQ ID NO: 1518 -20.4 -35.7 84.1 -14.1 -1.1 -6.4
TGCTTACTGAGCCTTTTGGA
327 SEQ ID NO: 1519 -20.4 -35.9 86.9 -13.7 -1.4 -11.4
TCCCATTGAGAGTGAAACTG
374 SEQ ID NO: 1520 -20.4 -35 83.9 -14.6 0 -5.1
CCATATACAGTCCCATTGAG
384 SEQ ID NO: 1521 -20.4 -34.4 83.4 -14 0 -3.4
TCTTCTCCCGCCAGAGGAGA
808 SEQ ID NO: 1522 -20.4 -42.8 93.4 -15 -7.4 -20.1
TTTCTTTGTTTTTCGAGCTT
1598 SEQ ID NO: 1523 -20.4 -27.6 74.6 -7.2 0 -5.7
AGTGGCCTGCTGAATTCCTT
1625 SEQ ID NO: 1524 -20.4 -38.1 89 -16.3 0 -107
GCATATAACACTTCAGGTTC
1750 SEQ ID NO: 1525 -20.4 -32 80.6 -11.6 0 -4.6
TTTGGTCATCCAGGTGTAAG
1890 SEQ ID NO: 1526 -20.4 -34.9 84.9 -12 -2.5 -8.2
TCTGCTCATTAATAATCAGA
2010 SEQ ID NO: 1527 -20.4 -30.1 76.7 -9 0 -9
TTCTCTGCTCATTAATAATC
2013 SEQ ID NO: 1528 -20.4 -28.9 75.1 -8.5 0 -2.5
GCAGTAAGGTTTTCATACAG
2124 SEQ ID NO: 1529 -20.4 -31.5 80.9 -11.1 0 -5.1
AGCAGTAAGGTTTTCATACA
2125 SEQ ID NO: 1530 -20.4 -31.5 80.9 -11.1 0 -5.1
AGCAATAGTTAAGGAGATTT
2322 SEQ ID NO: 1531 -20.4 -29.4 77.2 -9 0 -2.7
TTTGATATTTCCATTTGAAT
2423 SEQ ID NO: 1532 -20.4 -24.7 68.4 -4.3 0 -2.4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CAACAGATGAAAACAATAAA
2565 SEQ ID NO: 1533 -20.4 -25.4 70.7 -5 0 -2.5
CTCAACTGCTTCTGTTGCCA
2632 SEQ ID NO: 1534 -20.4 -36.8 87.3 -14.6 -1.8 -5.8
CGTCCACATATTAAGGTTTC
2720 SEQ ID NO: 1535 -20.4 -31 79.5 -10.6 0 -1.8
TCAGGTTTCCATGCATAAAT
2900 SEQ ID NO: 1536 -20.4 -31.6 79.6 -11.2 0 -7
TTTTTCAGGTTTCCATGCAT
2904 SEQ ID NO: 1537 -20.4 -31 79.1 -10.6 0 -5.3
TTGGTAGTTATTTTTTAAAT
3367 SEQ ID NO: 1538 -20.4 -22 66.2 -1.5 0 -2.7
TTACAGAAAGTTGGTAAGGT
3846 SEQ ID NO: 1539 -20.4 -31.3 81.3 -10.9 0 -4.5
AGGTTATCCATCAGCATTTC
4179 SEQ ID NO: 1540 -20.4 -33.1 81.2 -11.5 -1.1 -5.2
ACTGCTGGGGGAGGGCTGTA
4523 SEQ ID NO: 1541 -20.4 -45.3 98.6 -23 -1.9 -9
TAAACTCTTGTTGTAGGATA
4653 SEQ ID NO: 1542 -20.4 -29.7 78.2 -9.3 0 -6.4
GTCCTCATTTTATTTGGAAA
4673 SEQ ID NO: 1543 -20.4 -28 747 -6 -1.6 -6.6
TCTCCAAAATACTGAAAATA
4734 SEQ ID NO: 1544 -20.4 -27.2 73.4 -6.8 0 -0.4
GGTTTTATAGAAGTCCATCA
209 SEQ ID NO: 1545 -20.3 -30.9 78.9 -9.9 -0.4 -8.6
CCAAAAGTCTTCGCTGCTTG
300 SEQ ID NO: 1546 -20.3 -34.6 84.8 -13.7 -0.3 -5.2
GTCATTTCCCATCACTTTTG
416 SEQ ID NO: 1547 -20.3 -31.3 79 -11 0 0
ATTGAGGTTTGCAATGCTTT
503 SEQ ID NO: 1548 -20.3 -30.8 79.2 -8.8 -1.1 -11.3
CACTGGTCGACCTATTGAGG
516 SEQ ID NO: 1549 -20.3 -37.9 88 -13.4 -0.9 -16.5
TGAACTCTTGGGGTTCTCTG
539 SEQ ID NO: 1550 -20.3 -37.2 87.3 -11.4 -4.9 -19
ATACAATTTCACATTGCCAC
665 SEQ ID NO: 1551 -20.3 -30.3 78 -10 0 -2.2
ACAGTTTTCATCTATCAACA
782 SEQ ID NO: 1552 -20.3 -29.4 76.2 -9.1 0 -1.5
TACATTACTGGGGCTTGACA
926 SEQ ID NO: 1553 -20.3 -36.3 87.5 -16 0 -5.9
AGAAATGGCAGACATTTTAT
1073 SEQ ID NO: 1554 -20.3 -29.2 76.2 -5.3 -3.6 -97
TAATGACATCCTGAAGCTTC
1411 SEQ ID NO: 1555 -20.3 -33.2 81.3 -11.8 0 -10.1
AGAAAACTTTACAGCTTCCA
1446 SEQ ID NO: 1556 -20.3 -31.1 80.4 -10.8 0 -4
TTTTCGATAGCGGCATGCTG
1553 SEQ ID NO: 1557 -20.3 -35.7 85.5 -11.8 -1.8 -15.4
TCGAGCTTCCAGGTTCATTC
1586 SEQ ID NO: 1558 -20.3 -36.8 85.9 -11.9 -4.6 -13 ,
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TCCTCCAAGTTGAGTCTGGA
1788 SEQ ID NO: 1559 -20.3 -38.7 88.8 -16.3 -1.8 -11.9
TGATCCTCCAAGTTGAGTCT
1791 SEQ ID NO: 1560 -20.3 -36.5 85.4 -15.5 -0.5 -8.2
CATGATCCTCCAAGTTGAGT
1793 SEQ ID NO: 1561 -20.3 -35.2 83.6 -14 -0.8 -8.2
CAGCATGTGTTTACATTGGT
2054 SEQ ID NO: 1562 -20.3 -32.3 81.5 -11.1 -0.2 -9.3
CATACAGAGATACTCTTCAT
2111 SEQ ID NO: 1563 -20.3 -31.4 78.4 -10.4 -0.5 -7.6
GAAGAGAGAAGCAGTAAGGT
2134 SEQ ID NO: 1564 -20.3 -35.3 85 -15 0 -27
TGACAATGGCTTTTCCTAGC
2220 SEQ ID NO: 1565 -20.3 -34.9 85 -13.6 -0.9 -6.8
TAACATCTCGGGGAATTCAA
2372 SEQ ID NO: 1566 -20.3 -33.4 82 -11.9 0 -10.4
TTTCAGCTAACATCTCGGGG
2379 SEQ ID NO: 1567 -20.3 -36.6 86.9 -15.8 0 -8.2
AAACTTTTTTTCAGGTTTCC
2910 SEQ ID NO: 1568 -20.3 -27 74.4 -5.6 -1 -4.4
TTATTTGGGAAAGCTACGAA
3086 SEQ ID NO: 1569 -20.3 -30.7 79.8 -10.4 0 -5
GATAGCTAAGTGCCATCAGG
3156 SEQ ID NO: 1570 -20.3 -367 86.7 -12.5 -3.9 -9.8
ATTTGAGTCAATTTTTGTGG
3182 SEQ ID NO: 1571 -20.3 -27.4 73.7 -6.6 0 -8.2
TAATTTGCACAACCTATATA
3272 SEQ ID NO: 1572 -20.3 -27.5 75 -7.2 0 -4.2
CAGTTAGGACTGTTAATTTG
3285 SEQ ID NO: 1573 -20.3 -29 77.4 -6.6 -1.9 -11.8
TTAGTGAATACCAATATGAT
4017 SEQ ID NO: 1574 -20.3 -27.8 73.9 -7.5 0 -1.5
ATTAGTGAATACCAATATGA
4018 SEQ ID NO: 1575 -20.3 -27.8 73.9 -7.5 0 -2.3
GATTAGTGAATACCAATATG
4019 SEQ ID NO: 1576 -20.3 -27.8 74.1 -7.5 0 -3
AGATTAGTGAATACCAATAT
4020 SEQ ID NO: 1577 -20.3 -27.8 74 -7.5 0 -3
CATTTCTTTGACCCCTACAA
4165 SEQ ID NO: 1578 -20.3 -32.4 81.8 -12.1 0 -1
TTTGGTTTCTGAGACCATCG
4237 SEQ ID NO: 1579 -20.3 -34.1 82.9 -77 -4.2 -20.4
CCTCTTCCCCTAGAGCAAAC
4259 SEQ ID NO: 1580 -20.3 -38.5 89.6 -15.5 -2.7 -7.8
TCTGTAATCTCACTGGGTCA
4323 SEQ ID NO: 1581 -20.3 -36.7 86.2 -16.4 0 -7
CCTCCTCTTAGGGTTTTATA
220 SEQ ID NO: 1582 -20.2 -33.1 83.5 -11.5 -1.3 -7.4
CAGTGAGGGTGAAGACGCAG
257 SEQ ID NO: 1583 -20.2 -39.8 90.3 -18.4 -1.1 -4.6
CCCCGAGGAAAGGCTGATTT
458 SEQ ID NO: 1584 -20.2 -38.5 89.4 -17.7 -0.3 -6.8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CTTCTCCCGCCAGAGGAGAA
807 SEQ ID NO: 1585 -20.2 -41.3 92 -15 -6.1 -17.5
TCGAGTTTCCTTCCAAAAGG
837 SEQ ID NO: 1586 -20.2 -33.8 83.4 -12.5 -1 -5.7
GTTACATTACTGGGGCTTGA
928 SEQ ID NO: 1587 -20.2 -35.1 85.9 -13.4 -1.4 -7.2
AAATCTTCTTTTTCTGTTTT
961 SEQ ID NO: 1588 -20.2 -23.8 67.9 -3.6 0 0
AACAGAAATGGCAGACATTT
1076 SEQ ID NO: 1589 -20.2 -31.1 79.1 -9.4 -1.4 -5
AGGCTTCTGATCCTGCTGTT
1160 SEQ ID NO: 1590 -20.2 -38.4 88.7 -16.6 -1.5 -9.2
GTTGAGGAGCTGGATGGAGG
1345 SEQ ID NO: 1591 -20.2 -41 91.1 -20.8 0 -5.3
TGTTGAGGAGCTGGATGGAG
1346 SEQ ID NO: 1592 -20.2 -39.8 89.8 -19.6 0 -5.3
AAGACTCCATAATGACATCC
1420 SEQ ID NO: 1593 -20.2 -33.4 81.1 -13.2 0 -3.5
GCTCTTTTGAAGAAAACTTT
1456 SEQ ID NO: 1594 -20.2 -27.3 74.2 -6.4 0 -9
TCCACTGCTCTTTTGAAGAA
1462 SEQ ID NO: 1595 -20.2 -33.6 827 -12.7 0 -9
GTTTTTCGAGCTTCCAGGTT
1591 SEQ ID NO: 1596 -20.2 -337 83.5 -13 -0.1 -6.6
TTTTTTTCTTTGTTTTTCGA
1602 SEQ ID NO: 1597 -20.2 -22.7 66.4 -2.5 0 -2.5
TCCTGCATATAACACTTCAG
1754 SEQ ID NO: 1598 -20.2 -33.1 82 -12.9 0 -4.5
TTCACTGCTGCAATCACTTG
1840 SEQ ID NO: 1599 -20.2 -34.6 83.9 -13.9 0 -7.8
TGCCCATTTCACTGCTGCAA
1847 SEQ ID NO:1600 -20.2 -37.9 88.9 -16.9 -0.6 -7.9
AACCTGGTATTGCCTTTGCC
1863 SEQ ID NO: 1601 -20.2 -36.8 88.5 -14.6 -2 -10.7
GAAACCTGGTATTGCCTTTG
1865 SEQ ID NO: 1602 -20.2 -33.4 83.5 -11.6 -1.1 -10.7
CCAGGTGTAAGTTCCTGAAA
1881 SEQ ID NO: 1603 -20.2 -34.7 85.3 -9.7 -4.8 -12.2
AAGAAACATCCAGGAGTACT
1922 SEQ ID NO: 1604 -20.2 -34 83.4 -13.2 0 -8.4
AGAAGCAGTAAGGTTTTCAT
2128 SEQ ID NO: 1605 -20.2 -31.3 80.1 -9.8 -1.2 -8.8
CCTAGCTCTTTGATGTAGGT
2206 SEQ ID NO: 1606 -20.2 -35.3 85.6 -13 -2.1 -9.9
ACCGCTGCCAGTTCTGGCTG
2256 SEQ ID NO: 1607 -20.2 -42.9 95.1 -16.4 -5.6 -20.6
TTTTGATATTTCCATTTGAA
2424 SEQ ID NO: 1608 -20.2 -24.5 68.5 -4.3 0 -2.4
TTTTTGATATTTCCATTTGA
2425 SEQ ID NO: 1609 -20.2 -24.5 68.5 -4.3 0 -2.4
GACAAACTGATAGTTTATAC
2520 SEQ ID NO: 1610 -20.2 -27.6 74.6 -6.4 -0.6 -9.7 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
AACAACAGATGAAAACAATA
2567 SEQ ID NO: 1611 -20.2 -26.7 72.6 -6.5 0 -2
CTACTCTCCCATCACTGAAA
2663 SEQ ID NO: 1612 -20.2 -35.4 84.6 -15.2 0 -4.6
ACTTTGGGCACTGGTGGTTT
2771 SEQ ID NO: 1613 -20.2 -37.5 897 -15.3 0 -12.2
TTGGGAAAGCTACGAACTAG
3082 SEQ ID NO: 1614 -20.2 -34.2 84.9 -14 0 -5.3
GTCTTCTGATAGCTAAGTGC
3163 SEQ ID NO: 1615 -20.2 -34.5 83.8 -13.6 -0.4 -7.5
CACTGGACTAGGTGCTCTAT
3307 SEQ ID NO: 1616 -20.2 -37.8 88.5 -15.4 -2.2 -8.9
CAGCTTTCTTGCCATATAGC
3425 SEQ ID NO: 1617 -20.2 -34 83.8 -12.7 -1 -5.7
GTTTACCAGCTTTCTTGCCA
3431 SEQ ID NO: 1618 -20.2 -34.5 85.4 -13.2 -1 -4.2
GTCTCTCAGCTGTGTTACAG
3520 SEQ ID NO: 1619 -20.2 -36.4 86.2 -14.7 -0.7 -10.9
AAATTCACAGGACTTGTGTT
3767 SEQ ID NO: 1620 -20.2 -31 79.3 -7.6 -1.4 -14.6
CCAGTCTCCATCTCCCTCTT
4273 SEQ ID NO: 1621 -20.2 -40 89.8 -19.8 0 -1.4
ACCAGTCTCCATCTCCCTCT
4274 SEQ ID NO: 1622 -20.2 -41.3 91.3 -21.1 0 -1.3
ATTTTATTTGGAAATAAACT
4667 SEQ ID NO: 1623 -20.2 -22 65.2 -1.8 0 -7.3
CATTTTATTTGGAAATAAAC
4668 SEQ ID NO: 1624 -20.2 -22 65.8 -1.8 0 -7.3
TCCATCACATCTCCCCTCTC
196 SEQ ID NO: 1625 -20.1 -40.5 89.3 -20.4 0 0
CTTCACAGTAGCTCCTCCTC
233 SEQ ID NO: 1626 -20.1 -38.9 89.2 -18.8 0 -6
AGTCTGTTTCCCCCGAGGAA
468 SEQ ID NO: 1627 -20.1 -39.9 91.2 -18.1 -1.7 -7.5
TCTTGGGGTTCTCTGGAACA
534 SEQ ID NO: 1628 -20.1 -38.4 89.1 -14.9 -3.4 -13
ATGTTGCTGTTCTGAAGATA
620 SEQ ID NO: 1629 -20.1 -31.8 79.6 -11.7 0 -5.3
AAAGGTGCTTTGGTCTGTGG
686 SEQ ID NO: 1630 -20.1 -36.5 87.7 -15.5 0 -9.6
ATCAACAGGTCTGATCTCCA
769 SEQ ID NO: 1631 -20.1 -36.7 85.1 -15 -1.1 -10.8
CTGCTGTTGAGAAAGGGATG
1148 SEQ ID NO: 1632 -20.1 -35.9 85.4 -15.1 -0.5 -3.8
GACATCCTGAAGCTTCATCA
1407 SEQ ID NO: 1633 -20.1 -35.5 83.5 -11.8 0 -15.3
TGACATCCTGAAGCTTCATC
1408 SEQ ID NO: 1634 -20.1 -35.5 83.5 -11.8 0 -15.3
ACTCCATAATGACATCCTGA
1417 SEQ ID NO: 1635 -20.1 -34.6 82.6 -14.5 0 -4.6
AGCTCTTTGATGTAGGTCAT
2203 SEQ ID NO: 1636 -20.1 -34.2 83 -12.5 -0.9 -10.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecul Site oligo binding formation Duplex structure oligo oligo
TTCCTAGCTCTTTGATGTAG
2208 SEQ ID NO: 1637 -20.1 -33.1 82.3 -13 0 -3.6
ATTCAATACTCATGGTCTTA
2358 SEQ ID NO: 1638 -20.1 -29.9 76.8 -9.8 0 -5.2
CAGGACAAACTGATAGTTTA
2523 SEQ ID NO: 1639 -20.1 -30.5 79.1 -8.1 -2.3 -9.7
AAACAACAGATGAAAACAAT
2568 SEQ ID NO: 1640 -20.1 -26.3 71.8 -6.2 0 -2.5
GTCCACATATTAAGGTTTCT
2719 SEQ ID NO: 1641 -20.1 -30.7 79 -10.6 0 -2.1
GATTACGTCCACATATTAAG
2725 SEQ ID NO: 1642 -20.1 -29.2 76.6 -9.1 0 -3
TGAAGCAGATATATACAAAA
3243 SEQ ID NO: 1643 -20.1 -27.6 74 -7.5 0 -3.5
ATAATTCTCCACTGAAGCAG
3255 SEQ ID NO: 1644 -20.1 -32.9 81.3 -12.1 -0.5 -5.5
ACCTATATAATTCTCCACTG
3261 SEQ ID NO: 1645 -20.1 -31.2 79.4 -11.1 0 -3.4
ATTTGCACAACCTATATAAT
3270 SEQ ID NO: 1646 -20.1 -27.3 74 -7.2 0 -4.4
TTGCAAAAATAGGGCGTTAG
3403 SEQ ID NO: 1647 -20.1 -31.4 81.6 -10.8 -0.1 -6.5
TGCCATATAGCCATTGCAAA
3416 SEQ ID NO: 1648 -20.1 -33.9 83.8 -12.9 -07 -67
AATAATTTGGCCACCTTGAA
3913 SEQ ID NO: 1649 -20.1 -31.1 80.1 -9.4 0 -11.1
ATAACTTCCTCTGTAATCTC
4332 SEQ ID NO: 1650 -20.1 -31.2 78.8 -11.1 0 -0.3
ACCTTCCTGTCTCCTGTTTA
4455 SEQ ID NO: 1651 -20.1 -36.2 86.9 -16.1 0 0
GTGAAAGTAACCCGCTATTA
4502 SEQ ID NO: 1652 -20.1 -32.2 82.4 -11.6 -0.2 -3.4
GTACCTCTATGCAAACTATT
4574 SEQ ID NO: 1653 -20.1 -30.9 79.8 -10.8 0 -4.9
TAGCTCCTCCTCTTAGGGTT
225 SEQ ID NO: 1654 -20 -38.9 90.7 -16.6 -2.3 -7.6
AAGCGACAGCCAGTGAGGGT
267 SEQ ID NO: 1655 -20 -42 93.9 -20.4 -1.6 -7.2
GGGGGCAGCAGACACAGCAG
566 SEQ ID NO: 1656 -20 -45 96.9 -24 -0.9 -4.9
TGGGGGCAGCAGACACAGCA
567 SEQ ID NO: 1657 -20 -45 97.5 -24 -0.9 -6.2
CTCTGTGGGGGCAGCAGACA
572 SEQ ID NO: 1658 -20 -44 95.8 -19.9 -4.1 -12.8
CTGTGGTATACAATTTCACA
672 SEQ ID NO: 1659 -20 -30.5 78.8 -8.3 -2.2 -8
AGTGTTACATTACTGGGGCT
931 SEQ ID NO: 1660 -20 -36.1 87.7 -14.6 -1.4 -7.2
ATTACCCCAGGGGTGCAGAG
988 SEQ ID NO: 1661 -20 -41.8 94 -14.9 -4.3 -21.9
TAAACTGTGCCCAGTTTCTC
1015 SEQ ID NO: 1662 -20 -34.8 85.3 -11.1 -3.4 -15 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
CTGACAGTAAACTGTGCCCA 1022 SEQ ID NO: 1663 -20 -37 88.2 -13.7 -3.3 -12.1 ATCCTGAAGCTTCATCAGAG
1404 SEQ ID NO: 1664 -20 -35.4 83.5 -11.8 -2.4 -15.3 CATCCTGAAGCTTCATCAGA
1405 SEQ ID NO: 1665 -20 -35.4 83.4 -11.8 -2.4 -15.2 CATTCCTTCCAGCACATAGG
1497 SEQ ID NO: 1666 -20 -36.2 86 -16.2 0 -57
GTGGCCTGCTGAATTCCTTT 1624 SEQ ID NO: 1667 -20 -36.9 87.4 -16.3 0 -8.5
CTGCATATAACACTTCAGGT 1752 SEQ ID NO: 1668 -20 -32.9 82.2 -12.9 0 -4.5
GCAGTAGGGTCATTTGGTCA 1902 SEQ ID NO: 1669 -20 -37.4 88 -15.4 -2 -10.8
CAGTAAGGTTTTCATACAGA 2123 SEQ ID NO: 1670 -20 -30.5 79.1 -10.5 0 -5.1
AACAGATGAAAACAATAAAA 2564 SEQ ID NO: 1671 -20 -24.2 68.6 -4.2 0 -2.5
AGTGAGAGGAATTACTTTGT 3550 SEQ ID NO: 1672 -20 -31.6 80 -97 -1.9 -6.4
GAAGAAATTCACAGGACTTG 3771 SEQ ID NO: 1673 -20 -31.4 79.1 -8.9 -2.5 -9.7
TACCTCTATGCAAACTATTG 4573 SEQ ID NO: 1674 -20 -30.8 79.7 -10.8 0 -4.9
ATAGAAGTCCATCACATCTC 203 SEQ ID NO: 1675 -19.9 -33.7 80.9 -13.8 0 -3.5
CTTTCTTCCAAAAGCTTTAA 487 SEQ ID NO: 1676 -19.9 -27.3 75 -6.8 0 -8.6
GCAGACACAGCAGTGGATGC 559 SEQ ID NO: 1677 -19.9 -40.8 91 -14.1 -6.8 -18.9
TGGTATACAATTTCACATTG 669 SEQ ID NO: 1678 -19.9 -28.2 75.1 -8.3 0 -7.1
AAAATGAGAGGCTTGCAGTC 862 SEQ ID NO: 1679 -19.9 -34.8 84.1 -14.2 -0.3 -8.5
TGTCCGGTAAAATGAGAGGC 870 SEQ ID NO: 1680 -19.9 -36.7 87 -16.8 0 -6.3
CAGAAATGGCAGACATTTTA 1074 SEQ ID NO: 1681 -19.9 -30.2 78 -6.7 -3.6 -9.7
GGGAATTGGTGGAATGACAT 1187 SEQ ID NO: 1682 -19.9 -35.1 837 -14.5 -0.4 -5.6
AAGTTGTCATCTCCAGATCC 1237 SEQ ID NO: 1683 -19.9 -35.5 83.6 -15.6 0 -4.8
AGACTCCATAATGACATCCT 1419 SEQ ID NO: 1684 -19.9 -34.6 82.7 -14.7 0 -3.5
TGATGCAATCATTCCTTCCA 1506 SEQ ID NO: 1685 -19.9 -34.1 81.9 -14.2 0 -7.3
AGTTGAGTCTGGAACAGAGC 1781 SEQ ID NO: 1686 -19.9 -37.4 87.2 -15.9 -1.4 -10.6
TACAGCATGTGTTTACATTG 2056 SEQ ID NO: 1687 -19.9 -30.3 78.8 -9.6 0 -9.3
CTCTTCAGACCGTCCTTAGG 2158 SEQ ID NO: 1688 -19.9 -37.9 88.3 -17.5 -0.1 -7.4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target imolecular molecul Site oligo binding formation Duplex structure oligo oligo
TGGCTCTTCAGACCGTCCTT
2161 SEQ ID NO: 1689 -19.9 -40 90.9 -18.2 -1.9 -8.7
GCTAACATCTCGGGGAATTC
2374 SEQ ID NO: 1690 -19.9 -35.9 85 -14.6 0 -10.7
ACAAACTGATAGTTTATACA
2519 SEQ ID NO: 1691 -19.9 -27.3 74.3 -6.4 -0.6 -9.7
GGGGGGGATACACCAACAGA
2855 SEQ ID NO: 1692 -19.9 -42.1 94 -19.1 -3.1 -7.1
CCACTGAAGCAGATATATAC
3247 SEQ ID NO: 1693 -19.9 -32.5 81 -12.1 -0.1 -7.2
TTTGCACAACCTATATAATT
3269 SEQ ID NO: 1694 -19.9 -27.1 74.2 -7.2 0 -4.4
AATTTGCACAACCTATATAA
3271 SEQ ID NO: 1695 -19.9 -27.1 74.2 -7.2 0 -4.4
TTTTTAAATTAGTCTTTTGC
3356 SEQ ID NO: 1696 -19.9 -22.9 67.9 -3 0 -1.9
AGACAGGGCCTCTTGGTAGT
3379 SEQ ID NO: 1697 -19.9 -40.8 92.9 -18.1 -2.8 -10.9
CATTGCAAAAATAGGGCGTT
3405 SEQ ID NO: 1698 -19.9 -31.2 80.6 -10.8 -0.1 -6.7
TACCAGCTTTCTTGCCATAT
3428 SEQ ID NO: 1699 -19.9 -34 84 -13.2 -0.8 -4
AGAGGAATTACTTTGTCTGA
3546 SEQ ID NO: 1700 -19.9 -31.8 79.9 -9.9 -2 -8.4
TGACAGATGGGAATGTGAAA
3624 SEQ ID NO: 1701 -19.9 -34 82.1 -13 -1 -4.3
GTAGCTGAGCTTTCCTGTAC
3668 SEQ ID NO: 1702 -19.9 -36.5 87.3 -14.8 0 -11.8
TATACAAGCTATTTTACAAT
4086 SEQ ID NO: 1703 -19.9 -25.1 71.1 -5.2 0 -5
TGAGACCATCGCTGCCTGTA
4228 SEQ ID NO: 1704 -19.9 -40.4 91.1 -20.5 0 -4.5
CCTTCCTGTCTCCTGTTTAC
4454 SEQ ID NO: 1705 -19.9 -36.2 86.6 -16.3 0 0
TGTAAGCACCACCTTCCTGT
4465 SEQ ID NO: 1706 -19.9 -38.2 90.1 -18.3 0 -2.5
AGTGAGGGTGAAGACGCAGA
256 SEQ ID NO: 1707 -19.8 -40.1 90.7 -19.1 -1.1 -4.5
ATCAACCAAAAGTCTTCGCT
305 SEQ ID NO: 1708 -19.8 -32.7 81.5 -12.9 0 -3.2
CTGTTCTGAAGATACATCAG
614 SEQ ID NO: 1709 -19.8 -32 79.4 -11.1 -1 -8
GGTGCCAGTCTGGCCCTTCA
641 SEQ ID NO: 1710 -19.8 -44 96 -15.1 -6.9 -26.4
TGGTGCCAGTCTGGCCCTTC
642 SEQ ID NO: 1711 -19.8 -44 96 -15.1 -6.9 -26.4
TTCTTTTTCTGTTTTCACTT
956 SEQ ID NO: 1712 -19.8 -26.1 72.1 -6.3 0 0
GCCTGACAGTAAACTGTGCC
1024 SEQ ID NO: 1713 -19.8 -38.3 89.6 -15.3 -3.2 -12
CATGAACAGAAATGGCAGAC
1080 SEQ ID NO: 1714 -19.8 -33.8 82.1 -12.5 -1.4 -6 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TGGTACATCTGTCCTCCAGA
1111 SEQ ID NO: 1715 -19.8 -39.1 89.2 -17.5 -1.8 -8.5
CCAGATCCTTGGCACCTATT
1225 SEQ ID NO: 1716 -19.8 -37.4 87.8 -16.2 -1.3 -6.8
GGGCTTGAATAGCCATTAGA
1300 SEQ ID NO: 1717 -19.8 -35.4 85.5 -9.9 -5.7 -14.1
ACATCCTGAAGCTTCATCAG
1406 SEQ ID NO: 1718 -19.8 -35.2 83.5 -11.8 -1 -15.3
ATAATGACATCCTGAAGCTT
1412 SEQ ID NO: 1719 -19.8 -31.9 79.5 -12.1 0 -5.1
TAGTGGCCTGCTGAATTCCT
1626 SEQ ID NO: 1720 -19.8 -38.5 89.6 -16.3 -1 -12.9
TCAGAGGTTTCTTGTGAGAC
1651 SEQ ID NO: 1721 -19.8 -35.2 84.1 -12.5 -2.9 -11.1
TTTGCCCATTTCACTGCTGC
1849 SEQ ID NO: 1722 -19.8 -36.7 87.4 -16.9 0 -2.3
GGTAGAGTCATTCTCTGCTC
2023 SEQ ID NO: 1723 -19.8 -37 85.9 -9.5 -7.7 -18.4
GGCTCTTCAGACCGTCCTTA
2160 SEQ ID NO: 1724 -19.8 -39.2 90 -17.5 -1.9 -8.9
AACCGCTGCCAGTTCTGGCT
2257 SEQ ID NO: 1725 -19.8 -41.7 94.2 -15.6 -5.6 -20.6
TTGGTATCTGATTGGTGATG
2400 SEQ ID NO: 1726 -19.8 -33.1 81 -12.4 -0.8 -5.4
TAAAACAAAACAACAGATGA
2575 SEQ ID NO: 1727 -19.8 -26.5 72.7 -67 0 -2.5
CATCCTTCTTTGACTGTGGA
2745 SEQ ID NO: 1728 -19.8 -35 83.9 -14.3 -0.8 -5.2
AGAGTTTATTTGGGAAAGCT
3091 SEQ ID NO: 1729 -19.8 -31.2 80.3 -10.4 -0.9 -8.4
TTTTTTTTGACAAGAATACT
3206 SEQ ID NO: 1730 -19.8 -24 68.8 -3.7 0 -7.8
TGCACAACCTATATAATTCT
3267 SEQ ID NO: 1731 -19.8 -29.8 77.8 -10 0 -3.5
GACTGTTAATTTGCACAACC
3278 SEQ ID NO: 1732 -19.8 -31.2 80.2 -10.9 -0.1 -5.4
GGACTGTTAATTTGCACAAC
3279 SEQ ID NO: 1733 -19.8 -31.2 80.2 -10.9 -0.1 -5.4
CTTGGTAGTTATTTTTTAAA
3368 SEQ ID NO: 1734 -19.8 -23 68.6 -3.2 0 -27
CCATTGCAAAAATAGGGCGT
3406 SEQ ID NO: 1735 -19.8 -33.6 83.9 -12.9 -0.7 -6.7
TTTGTCTGATTAAAAGTCTC
3535 SEQ ID NO: 1736 -19.8 -28.2 74.7 -8.4 0 -3.7
GTGGCTTAGTAAATATGTTA
3876 SEQ ID NO: 1737 -19.8 -28.6 77.2 -8.8 0 -5.1
AAGCTATTTTACAATCATTT
4081 SEQ ID NO: 1738 -19.8 -24.5 69.4 -47 0 -5
CTGTATGTGAAAGTAACCCG
4508 SEQ ID NO: 1739 -19.8 -33 83 -13.2 0 -1.7
TCTTGTTGTAGGATAGAAAG
4648 SEQ ID NO: 1740 -19.8 -30.7 79 -10.4 0 -8 _
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecu Site oligo binding formation Duplex structure oligo oligo
CATGTCCTCATTTTATTTGG
4676 SEQ ID NO: 1741 -19.8 -29.1 76.2 -9.3 0 -2.4
CGCCACCGTCCGCAGTTCCC
77 SEQ ID NO: 1742 -19.7 -45 97.1 -24.4 -0.7 -4.6
CCAGTGAGGGTGAAGACGCA
258 SEQ ID NO: 1743 -19.7 -41 92 -20.1 -1.1 -6.4
TCTGTCTCTCCCATATACAG
394 SEQ ID NO: 1744 -19.7 -36 85 -16.3 0 -4.1
TCTTGCTTAATTACCCCAGG
997 SEQ ID NO: 1745 -197 -34.9 86 -15.2 0 -5.2
CAGTAAACTGTGCCCAGTTT
1018 SEQ ID NO: 1746 -197 -34.3 85.4 -11.2 -3.3 -14.1
GACATTTTATTACCAATTAT
1063 SEQ ID NO: 1747 -19.7 -23.9 68.5 -4.2 0 -0.1
CCATGAACAGAAATGGCAGA
1081 SEQ ID NO: 1748 -19.7 -34.9 83.7 -12.5 -2.7 -7.5
TCCTTGGCACCTATTCCAAT
1220 SEQ ID NO: 1749 -19.7 -36.2 86.5 -15 -1.4 -6.5
TGAAGAAAACTTTACAGCTT
1449 SEQ ID NO: 1750 -19.7 -28.7 76.8 -8.1 -0.8 -6.9
TTCCAGCACATAGGTAATTG
1491 SEQ ID NO: 1751 -19.7 -32.8 82.3 -13.1 0 -2.5
TGTAGTGGCCTGCTGAATTC
1628 SEQ ID NO: 1752 -19.7 -37.4 87.9 -14.9 -2.2 -13.6
GGGGTGAGTTGTGGTAACGT
1702 SEQ ID NO: 1753 -19.7 -38.6 90.5 -18.9 0 -5.4
AGGGGTGAGTTGTGGTAACG
1703 SEQ ID NO: 1754 -197 -38.5 90.4 -18.8 0 -5.4
CCTGCATATAACACTTCAGG
1753 SEQ ID NO: 1755 -197 -34 83.7 -12.9 -1.3 -7.5
TCATTCTCTGCTCATTAATA
2016 SEQ ID NO: 1756 -197 -30.1 767 -10.4 0 -2.5
CTTGGCTCTTCAGACCGTCC
2163 SEQ ID NO: 1757 -19.7 -40 90.5 -18.4 -1.9 -9.4
ATCCAAGAGTTTTGTCAGTT
2282 SEQ ID NO: 1758 -197 -31.2 79.3 -11 0 -8
ATCTCGGGGAATTCAATACT
2368 SEQ ID NO: 1759 -19.7 -33.6 81.8 -12.5 0 -10.7
ATTTCCATTTGAATATTTTG
2417 SEQ ID NO: 1760 -197 -23.2 66.6 -3.5 0 -4.1
AACAAAACAACAGATGAAAA
2572 SEQ ID NO: 1761 -197 -26.1 71.9 -6.4 0 -2.5
TCTAGAAAATTTCATCCAGC
2832 SEQ ID NO: 1762 -19.7 -30.7 78.1 -11 0 -6.8
CTATACAGGGGGGGGATACA
2863 SEQ ID NO: 1763 -197 -40.3 92.4 -20.6 0 -0.8
CCAGTTAGGACTGTTAATTT
3286 SEQ ID NO: 1764 -19.7 -30.2 79.3 -8.1 -2.2 -12.5
ACCAGTTAGGACTGTTAATT
3287 SEQ ID NO: 1765 -19.7 -31.5 81.1 -9.4 -2.2 -12.5
TTAGGTACAGACAGGGCCTC
3387 SEQ ID NO: 1766 -19.7 -40 92 -18.4 -1.9 -107 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
AAAATAGGGCGTTAGGTACA 3398 SEQ ID NO: 1767 -19.7 -33.1 84.3 -13.4 0 -7.2
ACCAGCTTTCTTGCCATATA 3427 SEQ ID NO: 1768 -19.7 -34 84 -13.2 -1 -4
AGTCTCTCAGCTGTGTTACA 3521 SEQ ID NO: 1769 -197 -36.4 86.4 -15.2 0 -10.9
CATTTTTGCCATATTAGAGA 3591 SEQ ID NO: 1770 -19.7 -28.3 75.1 -8.6 0 -1.2
TGTGGCTTAGTAAATATGTT 3877 SEQ ID NO: 1771 -19.7 -29.4 78.1 -9.7 0 -5
TTCTTGTGGCTTAGTAAATA 3881 SEQ ID NO: 1772 -19.7 -29.4 78.2 -9.7 0 -5
GTGAATACCAATATGATATA 4014 SEQ ID NO: 1773 -19.7 -27.2 72.9 -7.5 0 -4.8
GACCAGTCTCCATCTCCCTC 4275 SEQ ID NO: 1774 -19.7 -41.6 90.8 -21.9 0 -6.4
CTGAAGCTTCTGTTGTCATT 4545 SEQ ID NO: 1775 -19.7 -32.4 80.7 -10.5 -1.3 -12.5
TGCTGTGGGAATCCCAGGTC 433 SEQ ID NO: 1776 -19.6 -42 92.9 -16.7 -4.2 -19.5
GTTGCTGTTCTGAAGATACA 618 SEQ ID NO: 1777 -19.6 -32.9 81.5 -13.3 0 -6.1
TCATAGTGGTACATCTGTCC 1117 SEQ ID NO: 1778 -19.6 -35.9 85.1 -15.4 -0.3 -9.3
GATCCTTGGCACCTATTCCA 1222 SEQ ID NO: 1779 -19.6 -37.7 87.7 -18.1 0 -3.9
CACATAGGTAATTGTGCTGT 1485 SEQ ID NO: 1780 -19.6 -32.7 82.2 -10.8 -2.3 -6.8
TCCTTCCAGCACATAGGTAA
1494 SEQ ID NO: 1781 -19.6 -36.5 87.4 -16.2 -0.4 -6.9 TTCCTTCCAGCACATAGGTA
1495 SEQ ID NO: 1782 -19.6 -36.5 87.4 -16.2 -0.4 -6.9 CGTTGCAGGAACTATTGTTT
1685 SEQ ID NO: 1783 -19.6 -31.4 80.7 -10.9 -0.7 -8.3
AGAAACATCCAGGAGTACTG 1921 SEQ ID NO: 1784 -19.6 -35.2 847 -14.2 -1.3 -87
CATTCTCTGCTCATTAATAA 2015 SEQ ID NO: 1785 -19.6 -28.6 75 -9 0 -2.5
TAGAGTCATTCTCTGCTCAT 2021 SEQ ID NO: 1786 -19.6 -34.7 82.6 -11.6 -3.5 -10
AGGGTAGAGTCATTCTCTGC 2025 SEQ ID NO: 1787 -19.6 -37.9 87.6 -12.3 -6 -15
TCGTACATGCAGGGTAGAGT 2035 SEQ ID NO: 1788 -19.6 -38.2 89.2 -17.5 0 -10.1
ATAGTTAAGGAGATTTTCAA 2318 SEQ ID NO: 1789 -19.6 -27.2 73.5 -6.7 0 -9.8
TGATTTCAGCTAACATCTCG 2382 SEQ ID NO: 1790 -19.6 -32.3 80 -12.7 0 -6.1
CATGTAGTGCGTATTTAAAA 2590 SEQ ID NO: 1791 -19.6 -27.3 74.9 -6.6 0 -10.2
AAAGTGATGACGACTCAACT 2645 SEQ ID NO: 1792 -19.6 -33.1 81.2 -13.5 0 -5.5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CACTGGTGGTTTAGGTGCCA
2763 SEQ ID NO: 1793 -19.6 -39.1 91.5 -16.5 -3 -11.3
GTTAATTTGCACAACCTATA
3274 SEQ ID NO: 1794 -19.6 -28.2 76.4 -8.6 0 -4.4
AATAGGGCGTTAGGTACAGA
3396 SEQ ID NO: 1795 -19.6 -35.8 87.3 -16.2 0 -7.2
TGAAGAAATTCACAGGACTT
3772 SEQ ID NO: 1796 -19.6 -31.4 79.1 -8.9 -2.9 -10.3
ATCGCTGCCTGTATGAATGC
4221 SEQ ID NO: 1797 -19.6 -36.9 86.2 -17.3 0 -5.7
TACATACTTTACATACTTTA
4437 SEQ ID NO: 1798 -19.6 -25.4 72.1 -5.8 0 0
TCCTGTCTCCTGTTTACATA
4451 SEQ ID NO: 1799 -19.6 -34.4 84 -14.8 0 -4.4
TCCGCAGTTCCCGCCGCAGC
69 SEQ ID NO: 1800 -19.5 -46.2 98.5 -24.7 -2 -6.7
CTCCTCCTCTTAGGGTTTTA
222 SEQ ID NO: 1801 -19.5 -35.2 86.1 -14 -1.7 -7.4
GTATACAATTTCACATTGCC
667 SEQ ID NO: 1802 -19.5 -29.5 77.2 -10 0 -7.4
GCTGTTGAGGAGCTGGATGG
1348 SEQ ID NO: 1803 -19.5 -40.8 91.1 -19.4 -1.5 -11.7
AATGACATCCTGAAGCTTCA
1410 SEQ ID NO: 1804 -19.5 -34 82.1 -11.8 0 -13.5
TTGAAGAAAACTTTACAGCT
1450 SEQ ID NO: 1805 -19.5 -28.7 76.8 -8.1 -1 -5.3
GATGATGCAATCATTCCTTC
1508 SEQ ID NO: 1806 -19.5 -32.2 78.5 -11 -1.4 -11.1
TCGATGATGCAATCATTCCT
1510 SEQ ID NO: 1807 -19.5 -33.7 80.4 -12 -17 -12.4
GGGTCATTTGGTCATCCAGG
1896 SEQ ID NO: 1808 -19.5 -38.5 88.3 -16.6 -2.4 -10.8
GAGAGAAGCAGTAAGGTTTT
2131 SEQ ID NO: 1809 -19.5 -32.6 82 -11.9 -1.1 -7.8
GCCAGTTCTGGCTGGAGTTT
2250 SEQ ID NO: 1810 -19.5 -39.2 90.6 -13.2 -6.3 -20.6
CCGCTGCCAGTTCTGGCTGG
2255 SEQ ID NO: 1811 -19.5 -44 96.2 -18 -5.8 -21
CCATTTGAATATTTTGGTAT
2413 SEQ ID NO: 1812 -19.5 -25.8 71.4 -5.6 -0.5 -7.4
AGTTTTTTGATATTTCCATT
2428 SEQ ID NO: 1813 -19.5 -24.3 68.6 -4.8 0 -2.3
AAGTTTTTTGATATTTCCAT
2429 SEQ ID NO: 1814 -19.5 -24.3 68.6 -4.8 0 -2.3
ACTGGACTAGGTGCTCTATA
3306 SEQ ID NO: 1815 -19.5 -37 87.9 -167 -0.3 -8.9
TTACCAGCTTTCTTGCCATA
3429 SEQ ID NO: 1816 -19.5 -33.8 84.3 -13.2 -1 -37
TTGTCTGATTAAAAGTCTCT
3534 SEQ ID NO: 1817 -19.5 -29.4 76.4 -9.9 0 -3.7
AGATTAACCAATTGGTGACA
3639 SEQ ID NO: 1818 -19.5 -31.5 79.9 -8.3 -2.2 -15.6 . . -
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecu! Site oligo binding formation Duplex structure oligo oligo
AGCTTTCCTGTACCATCAGG
3661 SEQ ID NO: 1819 -19.5 -37.3 88 -14.1 -3.7 -10.1
TGTGTTAAACTCTATGGCAC
3753 SEQ ID NO: 1820 -19.5 -32.8 82.5 -10.6 -2.7 -8.2
GAGTTTACAGAAAGTTGGTA
3850 SEQ ID NO: 1821 -19.5 -30.4 79.4 -10.9 0 -3.2
TATTTTACACTATACAAGCT
4096 SEQ ID NO: 1822 -19.5 -27.4 75.2 -7.9 0 -3.1
ATATAGGTTATCCATCAGCA
4183 SEQ ID NO: 1823 -19.5 -32.6 80.9 -12.1 -0.9 -5
TCGCTGCCTGTATGAATGCA
4220 SEQ ID NO: 1824 -19.5 -37.9 88 , -17.6 -0.6 -7.8
CTAGAGCAAACTGTTTGGTT
4250 SEQ ID NO: 1825 -19.5 -32.1 82 -10.7 -1.1 -11.9
CTCAGCAACCTTCACTGCAC
4299 SEQ ID NO: 1826 -19.5 -38.1 88.6 -16.3 -2.3 -5.9
CATACTTTAGTGCAAGGGGA
4426 SEQ ID NO: 1827 -19.5 -35 85.8 -14.9 -0.1 -8.4
ATGTGAAAGTAACCCGCTAT
4504 SEQ ID NO: 1828 -19.5 -33.2 83 -13.2 -0.2 -3.4
ATTTACATATTGCTGGTACC
4589 SEQ ID NO: 1829 -19.5 -30.8 797 -10.8 0 -8.2
TGAGGGTGAAGACGCAGAAA
254 SEQ ID NO: 1830 -19.4 -37.6 87.9 -17 -1.1 -4.5
GAGAAGCGACAGCCAGTGAG
270 SEQ ID NO: 1831 -19.4 -40.1 90.2 -19.8 -0.8 -7.1
CAAAAGTCTTCGCTGCTTGG
299 SEQ ID NO: 1832 -19.4 -34.6 84.8 -13.8 -1.3 -5.8
AGGTTTGCAATGCTTTCTTC
499 SEQ ID NO: 1833 -19.4 -32.1 81.1 -12.1 0 -8.6
AAGGTGCTTTGGTCTGTGGT
685 SEQ ID NO: 1834 -19.4 -37.8 89.4 -17.8 0 -8.4
ATTTTCGGAACCAACGGGAA
1202 SEQ ID NO: 1835 -19.4 -34.1 83.9 -11.6 -3.1 -8.4
CATAATGACATCCTGAAGCT
1413 SEQ ID NO: 1836 -19.4 -33.1 81 -13.7 0 -3.9
ATTCCTTCCAGCACATAGGT
1496 SEQ ID NO: 1837 -19.4 -36.3 86.4 -16.2 -0.4 -6.9
GCCTGCTGAATTCCTTTTAT
1621 SEQ ID NO: 1838 -19.4 -32.6 81.6 -13.2 0 -5.2
GGTAGGGGTGAGTTGTGGTA
1706 SEQ ID NO: 1839 -19.4 -39.8 92.2 -20.4 0 -27
ATCCTCCAAGTTGAGTCTGG
1789 SEQ ID NO: 1840 -19.4 -37.4 87 -17.1 -0.8 -8
CAGCTAACATCTCGGGGAAT
2376 SEQ ID NO: 1841 -19.4 -36.8 86.6 -16 0 -10.7
TATTTGGGAAAGCTACGAAC
3085 SEQ ID NO: 1842 -19.4 -32 81.5 -12.6 0 -5
GTCAATTTTTGTGGTCTTCT
3176 SEQ ID NO: 1843 -19.4 -30 77.6 -10.6 0 -2.4
TGTCTAGCCATTTTTGCCAT
3599 SEQ ID NO: 1844 -19.4 -33.6 83.3 -14.2 0 -5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecular Site oligo binding formation Duplex structure oligo oligo
TTTGAGTTTACAGAAAGTTG
3853 SEQ ID NO: 1845 -19.4 -27.5 74.9 -8.1 0 -2.4
AAATAATTTGGCCACCTTGA
3914 SEQ ID NO: 1846 -19.4 -31.1 80.1 -10.1 0 -11.1
TTGGTTTCTGAGACCATCGC
4236 SEQ ID NO: 1847 -19.4 -36.6 86 -11.1 -4.2 -20.4
GCAAACTGTTTGGTTTCTGA
4245 SEQ ID NO: 1848 -19.4 -32 81.2 -11.1 -1.1 -10.6
TCTCCATCTCCCTCTTCCCC
4269 SEQ ID NO: 1849 -19.4 -42.6 92.6 -23.2 0 0
TCCTCATTTTATTTGGAAAT
4672 SEQ ID NO: 1850 -19.4 -26.9 72.5 -6 -1.4 -5.9
CGTCCGCAGTTCCCGCCGCA
71 SEQ ID NO: 1851 -19.3 -45.3 97.7 -24.3 -1.7 -6.4
CAGAGGAGCCGCTCGCCCGC
94 SEQ ID NO: 1852 -19.3 -47.6 99.2 -24.7 -3.4 -14.6
TCCCCTCTCCTGAGCAAGCA
185 SEQ ID NO: 1853 -19.3 -43.2 94.6 -22.3 -1.6 -5.9
TTAGGGTTTTATAGAAGTCC
213 SEQ ID NO: 1854 -19.3 -30.8 80.2 -97 -1.6 -11
GCGACAGCCAGTGAGGGTGA
265 SEQ ID NO: 1855 -19.3 -43.5 94.7 -20.3 -3.9 -10
ATTGAGAAGCGACAGCCAGT
273 SEQ ID NO: 1856 -19.3 -37.6 87.6 -17.2 -1 -4.6
CTGCTGCGCATTGCTTACTG
338 SEQ ID NO: 1857 -19.3 -37.2 88.2 -16 -1 -11.9
ACATTACTGGGGCTTGACAA
925 SEQ ID NO: 1858 -19.3 -35.9 86.8 -16.6 0 -5.9
CTTAATTACCCCAGGGGTGC
992 SEQ ID NO: 1859 -19.3 -38.3 90.6 -12.1 -4.3 -21.9
CTGTCCTCCAGAGGTACTCA
1103 SEQ ID NO: 1860 -19.3 -40.1 90.7 -17.5 -3 -14.4
GGCTTCTGATCCTGCTGTTG
1159 SEQ ID NO: 1861 -19.3 -38.4 88.3 -17.8 -1.2 -8.6
TAGGCTTCTGATCCTGCTGT
1161 SEQ ID NO: 1862 -19.3 -38.8 89.3 -17.9 -1.5 -9.2
CTGTTCGACCAGGGAAGTTC
1275 SEQ ID NO: 1863 -19.3 -37.6 87.7 -16.2 -1.1 -12.3
TTGAGTCTGGAACAGAGCTA
1779 SEQ ID NO: 1864 -19.3 -36.5 86.5 -15 -2 -12
GAGATACTCTTCATAAGATA
2105 SEQ ID NO: 1865 -19.3 -29.6 75.8 -10.3 0 -5.6
ATACAGAGATACTCTTCATA
2110 SEQ ID NO: 1866 -19.3 -30.6 77.5 -10.4 -0.8 -7.9
AGACTTTGGGCACTGGTGGT
2773 SEQ ID NO: 1867 -19.3 -40.2 92.4 -19.2 0 -11.6
CTATACCAGTTAGGACTGTT
3291 SEQ ID NO: 1868 -19.3 -33.1 83.3 -11.4 -2.2 -12.5
TCTATACCAGTTAGGACTGT
3292 SEQ ID NO: 1869 -19.3 -34.6 84.9 -12.9 -2.2 -12.5
GGTAGTTATTTTTTAAATTA
3365 SEQ ID NO: 1870 -19.3 -21.2 65.3 -1.9 0 -1.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecu! Site oligo binding formation Duplex structure oligo oligo
GTCTGATTAAAAGTCTCTCA
3532 SEQ ID NO: 1871 -19.3 -30.9 78.1 -11.6 0 -3.7
AGTTTAGTGAGAGGAATTAC
3555 SEQ ID NO: 1872 -19.3 -30.8 79.2 -10.8 -0.4 -3.4
TAGGGATGTGTAGTTTTACA
3729 SEQ ID NO: 1873 -19.3 -31.9 817 -11.7 -0.8 -7.1
TATAGGTTATCCATCAGCAT
4182 SEQ ID NO: 1874 -19.3 -32.6 80.9 -12.1 -1.1 -5.2
TGTGAAAGTAACCCGCTATT
4503 SEQ ID NO: 1875 -19.3 -33 83.3 -13.2 -0.2 -3.4
TCATTTTATTTGGAAATAAA
4669 SEQ ID NO: 1876 -19.3 -22.2 65.1 -2.9 0 -5.8
GCTTACTGAGCCTTTTGGAA
326 SEQ ID NO: 1877 -19.2 -347 85.3 -13.7 -1.4 -11.4
GACAGATCTGGCTGCTGCGC
349 SEQ ID NO: 1878 -19.2 -42.5 92.6 -19.2 -1.5 -16.4
CTGCTGTGGGAATCCCAGGT
434 SEQ ID NO: 1879 -19.2 -417 93 -17.2 -4.1 -18.6
AACTTTACAGCTTCCACAAG
1442 SEQ ID NO: 1880 -19.2 -32.1 82 -12.9 0 -4
GTTCATTCCAGCCTGAAGAC
1574 SEQ ID NO: 1881 -19.2 -36.3 85.4 -16.3 -0.6 -6.8
ACACTTCAGGTTCAATAACC
1743 SEQ ID NO: 1882 -19.2 -32.6 81.8 -12.3 -1 -5.4
ATTTCACTGCTGCAATCACT
1842 SEQ ID NO: 1883 -19.2 -33.6 82.2 -13.9 0 -7.9
CTTCAGACCGTCCTTAGGAA
2156 SEQ ID NO: 1884 -19.2 -36.7 87.1 -15.9 -1.2 -11
CACTTCATGCATAGAATCCA
2297 SEQ ID NO: 1885 -19.2 -33.1 80.9 -12.9 0 -9.9
GCAATAGTTAAGGAGATTTT
2321 SEQ ID NO: 1886 -19.2 -28.2 75.5 -9 0 -3.2
TTCAATACTCATGGTCTTAT
2357 SEQ ID NO: 1887 -19.2 -29.9 76.8 -107 0 -5.2
TCTATAAACCACATGTAGTG
2601 SEQ ID NO: 1888 -19.2 -30.9 79.6 -10.6 -0.9 -9.4
ATATTAAGGTTTCTAATTTC
2713 SEQ ID NO: 1889 -19.2 -23.9 68.6 -4.7 0 -4
TCCACATATTAAGGTTTCTA
2718 SEQ ID NO: 1890 -19.2 -29.8 78 -10.6 0 -4
GGGGGGATACACCAACAGAA
2854 SEQ ID NO: 1891 -19.2 -39.7 91.2 -16.9 -3.6 -8.7
AGCCATTGCAAAAATAGGGC
3408 SEQ ID NO: 1892 -19.2 -34.5 85.1 -13.7 -1.5 -8.4
AGTTTACCAGCTTTCTTGCC
3432 SEQ ID NO: 1893 -19.2 -34.5 85.5 -14.2 -1 -4.4
AAGTTATACAAACTACTTCA
3472 SEQ ID NO: 1894 -19.2 -27.1 74.5 -7.1 -0.6 -3.9
GAAATTCACAGGACTTGTGT
3768 SEQ ID NO: 1895 -19.2 -32.5 80.9 -10 -1.5 -14.7
GAAAGTTGGTAAGGTGCACA
3841 SEQ ID NO: 1896 -19.2 -34.6 85.4 -13.9 0 -11
I 35 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
ATATATTAGAAGTTCCATAT 3967 SEQ ID NO: 1897 -19.2 -25.9 70.9 -6.7 0 -4.9
GTCAGAATGGGAGGCAGAGG 4352 SEQ ID NO: 1898 -19.2 -41 91.1 -20.9 -0.8 -4.2
CTTTACATACTTTAGTGCAA 4431 SEQ ID NO: 1899 -19.2 -28 76.4 -8.8 0 -6.3
CACCTTCCTGTCTCCTGTTT 4456 SEQ ID NO: 1900 -19.2 -37 87.5 -17.8 0 0
TTTATAGAAGTCCATCACAT 206 SEQ ID NO: 1901 -19.1 -29.9 76.8 -10.8 0 -4
CATTGAGAGTGAAACTGCTT 371 SEQ ID NO: 1902 -19.1 -32.4 807 -12.2 -1 -6.7
TTAAGTCTGTTTCCCCCGAG 471 SEQ ID NO: 1903 -19.1 -36.4 87.4 -17.3 0 -2.4
GTCCGGTAAAATGAGAGGCT 869 SEQ ID NO: 1904 -19.1 -36.7 87 -17.6 0 -6.3 CCATTATCCTTAATTTTGGG
895 SEQ ID NO: 1905 -19.1 -29.1 77.2 -8.4 -1.6 -7.5 TCCATTATCCTTAATTTTGG
896 SEQ ID NO: 1906 -19.1 -28.2 75.3 -8.4 -0.4 -5 CAGTTTCTCTTGCTTAATTA
1004 SEQ ID NO: 1907 -19.1 -28 75.5 -8.9 0 -27
ATCCTTGGCACCTATTCCAA 1221 SEQ ID NO: 1908 -19.1 -36.2 86.5 -16.2 -0.8 -4.7
CAAGTTGTCATCTCCAGATC 1238 SEQ ID NO: 1909 -19.1 -34.3 81.9 -15.2 0 -4.8
CACAAGTTAAGACTCCATAA 1428 SEQ ID NO: 1910 -19.1 -30.5 79.1 -11.4 0 -4.9
TCATTCCTTCCAGCACATAG 1498 SEQ ID NO: 1911 -19.1 -35.3 84.4 -16.2 0 -2.7
GATAGCGGCATGCTGGGCAG 1548 SEQ ID NO: 1912 -19.1 -42.4 93.5 -18.5 -2.6 -17.7
TTGTTTTTCGAGCTTCCAGG 1593 SEQ ID NO: 1913 -19.1 -33.6 83.4 -14.5 0 -6.6
GGAGTACTGCAGTAGGGTCA 1910 SEQ ID NO: 1914 -19.1 -40.1 91.8 -17 -0.2 -16.1
GTCTATATGATCTCCACCCC 1956 SEQ ID NO: 1915 -19.1 -37.4 86.4 -18.3 0 -4.2
ATTCTCTGCTCATTAATAAT 2014 SEQ ID NO: 1916 -19.1 -27.6 73.1 -8.5 0 -2.5
GTAGAGTCATTCTCTGCTCA 2022 SEQ ID NO: 1917 -19.1 -35.8 84.4 -9.5 -7.2 -17.4
TTGGCTCTTCAGACCGTCCT 2162 SEQ ID NO: 1918 -19.1 -40 90.9 -19 -1.9 -9.4
ACTTCATGCATAGAATCCAA 2296 SEQ ID NO: 1919 -19.1 -31.9 79.5 -11.8 0 -9.9
TTCAACCACTTCATGCATAG 2303 SEQ ID NO: 1920 -19.1 -32.7 81.3 -13.6 0 -7
AGATTTTCAACCACTTCATG 2308 SEQ ID NO: 1921 -19.1 -30.1 77.2 -11 0 -2.1
AGCTAACATCTCGGGGAATT 2375 SEQ ID NO: 1922 -19.1 -35.6 85.3 -15.1 0 -10.7
I36
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
ACAGGACAAACTGATAGTTT
2524 SEQ ID NO: 1923 -19.1 -31.4 80.1 -9.5 -2.8 -8
AAAACAACAGATGAAAACAA
2569 SEQ ID NO: 1924 -19.1 -26.1 71.9 -7 0 -2.5
GCTAAGTGCCATCAGGTTAG
3152 SEQ ID NO: 1925 -19.1 -36.3 87.1 -15.6 -1.5 -7.4
CTGATAGCTAAGTGCCATCA
3158 SEQ ID NO: 1926 -19.1 -35.5 85.1 -12.5 -3.9 -9.8
ATTGCAAAAATAGGGCGTTA
3404 SEQ ID NO: 1927 -19.1 -30.4 79.7 -10.8 -0.1 -6.7
TTGCCATATAGCCATTGCAA
3417 SEQ ID NO: 1928 -19.1 -33.9 83.8 -13.4 -1.3 -5.6
TTTACCAGCTTTCTTGCCAT
3430 SEQ ID NO: 1929 -19.1 -33.4 83.6 -13.2 -1 -4
AGAAGTTATACAAACTACTT
3474 SEQ ID NO: 1930 -19.1 -27.1 74.6 -7.1 -07 -4
CACCTTGAATAGAAATCAAA
3902 SEQ ID NO: 1931 -19.1 -28 74.5 -8.4 -0.2 -3.8
TCCTCTGTAATCTCACTGGG
4326 SEQ ID NO: 1932 -19.1 -37.8 87.6 -16.6 -2.1 -7.6
GCTGTATGTGAAAGTAACCC
4509 SEQ ID NO: 1933 -19.1 -34 84.2 -14.9 0 -3.6
GCAGAAGGAGCAGGAGGGAA
23 SEQ ID NO: 1934 -19 -42 93.1 -22.1 -07 -3.2
GTCCATCAGTGAATATCAAC
119 SEQ ID NO: 1935 -19 -32.3 79.4 -13.3 0 -6.4
AATGATTCTTTGGAGTCCAT
133 SEQ ID NO: 1936 -19 -31.7 78.8 -8.9 -3.8 -14.2
GGACAGATCTGGCTGCTGCG
350 SEQ ID NO: 1937 -19 -42.4 927 -19.3 -2 -16.4
CAGTCCCATTGAGAGTGAAA
377 SEQ ID NO: 1938 -19 -35 83.9 -14.6 -1.1 -10.3
CTGTGGGAATCCCAGGTCAT
431 SEQ ID NO: 1939 -19 -39.7 89.8 -15.9 -3.9 -177
CTGTGGGGGCAGCAGACACA
570 SEQ ID NO: 1940 -19 -43.8 95.9 -217 -2.7 -14
AAAATGTCAAAGGTGCTTTG
694 SEQ ID NO: 1941 -19 -29.4 77.6 -9.5 -0.4 -9.6
TGTTACATTACTGGGGCTTG
929 SEQ ID NO: 1942 -19 -34.8 85.9 -14.3 -1.4 -7.2
ACCATGAACAGAAATGGCAG
1082 SEQ ID NO: 1943 -19 -34.7 83.8 -12.5 -3.2 -87
TTTCACTGCTGCAATCACTT
1841 SEQ ID NO: 1944 -19 -33.4 82.5 -13.9 0 -7.9
AGATCAGGAGCAAAACACAG
1993 SEQ ID NO: 1945 -19 -34.8 83.9 -15.8 0 -5.2
ACCGTCCTTAGGAACTGAAG
2150 SEQ ID NO: 1946 -19 -36.5 87.2 -15.9 -1.2 -11
TCTTCAGACCGTCCTTAGGA
2157 SEQ ID NO: 1947 -19 -38.2 88.6 -17.6 -1.6 -9.3
CTTTGATGTAGGTCATTCTA
2199 SEQ ID NO: 1948 -19 -30.9 78.7 -10.3 -0.9 -10.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TCTTTGATGTAGGTCATTCT
2200 SEQ ID NO: 1949 -19 -32 79.6 -11.4 -0.9 -10.9
TAGCTCTTTGATGTAGGTCA
2204 SEQ ID NO: 1950 -19 -34.4 83.9 -14.6 -0.3 -9.2
AGTTAAGGAGATTTTCAACC
2316 SEQ ID NO: 1951 -19 -30.3 78.5 -10.4 0 -9.8
CTAACATCTCGGGGAATTCA
2373 SEQ ID NO: 1952 -19 -34.6 83.4 -14.2 0 -10.7
ACAGATGAAAACAATAAAAA
2563 SEQ ID NO: 1953 -19 -24.2 68.6 -5.2 0 -2.5
GCACTGGTGGTTTAGGTGCC
2764 SEQ ID NO: 1954 -19 -40.4 92.8 -16.4 -5 -14.8
GTTAAAACCAGACAGTAATA
2971 SEQ ID NO: 1955 -19 -28.3 76.5 -9.3 0 -2.1
ACTACTTCAAAAGGTCCTGA
3461 SEQ ID NO: 1956 -19 -33.8 83.7 -14.1 0 -9
AAGAAGTTATACAAACTACT
3475 SEQ ID NO: 1957 -19 -27.1 74.6 -7.1 -0.9 -4.2
TTTTGAGTTTACAGAAAGTT
3854 SEQ ID NO: 1958 -19 -26.3 73 -6.3 -0.9 -3.6
TTGTGGCTTAGTAAATATGT
3878 SEQ ID NO: 1959 -19 -29.4 78.1 -10.4 0 -4.6
TCAAATTTCTTGTGGCTTAG
3887 SEQ ID NO: 1960 -19 -30 78.3 -11 0 -3.8
CCATCTCCCTCTTCCCCTAG
4266 SEQ ID NO: 1961 -19 -41.2 92 -22.2 0 -2
CTGACAGACTCACTGTTGGA
4384 SEQ ID NO: 1962 -19 -37.4 87 -11.9 -6.5 -15.4
TGTATGTGAAAGTAACCCGC
4507 SEQ ID NO: 1963 -19 -34.3 84.7 -15.3 0 -1.7
AACTGCTGGGGGAGGGCTGT
4524 SEQ ID NO: 1964 -19 -44.9 98 -24.3 -1.5 -9
TTGCTGGTACCTCTATGCAA
4580 SEQ ID NO: 1965 -19 -36.3 87.4 -13.6 -3.7 -13
GGCAACCTATGAGATTCTGC
4612 SEQ ID NO: 1966 -19 -36.6 85.9 -17.6 0 -5.5
TTGCTTACTGAGCCTTTTGG
328 SEQ ID NO: 1967 -18.9 -34.4 85.4 -13.7 -1.4 -11.4
ACTTTTGTTTCTGTCTCTCC
403 SEQ ID NO: 1968 -18.9 -32.5 ~ 81.1 -13.6 0 0
TCAACAGGTCTGATCTCCAA
768 SEQ ID NO: 1969 -18.9 -36.5 85.4 -16 -1.1 -11.1
TTCGAGTTTCCTTCCAAAAG
838 SEQ ID NO: 1970 -18.9 -31.4 80 -12.5 0 -4.5
GTGTCCGGTAAAATGAGAGG
871 SEQ ID NO: 1971 -18.9 -35.5 85.3 -16.6 0 -6.3
TCTCCATTATCCTTAATTTT
898 SEQ ID NO: 1972 -18.9 -27.3 73.3 -8.4 0 -0.4
AATCTTCTTTTTCTGTTTTC
960 SEQ ID NO: 1973 -18.9 -25.3 70.2 -6.4 0 0
CGATGAAATCTTCTTTTTCT
966 SEQ ID NO: 1974 -18.9 -27.2 72.4 -7.5 -0.6 -4.6 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TGAACAGAAATGGCAGACAT
1078 SEQ ID NO: 1975 -18.9 -33.8 82.2 -13.4 -1.4 -5.1
ATAGTGGTACATCTGTCCTC
1115 SEQ ID NO: 1976 -18.9 -35.9 85.2 -15.4 -1.5 -8.5
TGCTGTATTCATGTCATAGT
1130 SEQ ID NO: 1977 -18.9 -32.2 80.2 -12.1 -0.8 -10
CTTCCACAAGTTAAGACTCC
1432 SEQ ID NO: 1978 -18.9 -33.8 83.5 -14.9 0 -5
ATTTTCAGAGGTTTCTTGTG
1655 SEQ ID NO: 1979 -18.9 -29.9 77.5 -10.5 0 -7.6
ACTTCAGGTTCAATAACCTC
1741 SEQ ID NO:1980 -18.9 -32.8 81.8 -11.4 -2.5 -9.4
GCTCTTTGATGTAGGTCATT
2202 SEQ ID NO: 1981 -18.9 -33 81.4 -12.5 -0.9 -10.9
GACAATGGCTTTTCCTAGCT
2219 SEQ ID NO: 1982 -18.9 -34.9 85 -13.3 -2.7 -7.8
CCAGTTCTGGCTGGAGTTTC
2249 SEQ ID NO: 1983 -18.9 -38.2 88.8 -13.2 -6.1 -16.9
CAAGAGTTTTGTCAGTTGAT
2279 SEQ ID NO: 1984 -18.9 -30 77.4 -11.1 0 -7.3
AATCCAAGAGTTTTGTCAGT
2283 SEQ ID NO: 1985 -18.9 -31.2 79.3 -11.8 0 -8
CTTCATGCATAGAATCCAAG
2295 SEQ ID NO: 1986 -18.9 -31.8 79.3 -11.9 0 -9.9
TACAGGACAAACTGATAGTT
2525 SEQ ID NO: 1987 -18.9 -31.8 80.8 -10.1 -2.8 -6.5
TATATACAAAATATGAGCTT
3234 SEQ ID NO: 1988 -18.9 -25.5 71 -6.6 0 -5.5
TACCAGTTAGGACTGTTAAT
3288 SEQ ID NO: 1989 -18.9 -31.9 81.8 -10.6 -2.2 -12.5
CTCTTGGTAGTTATTTTTTA
3370 SEQ ID NO: 1990 -18.9 -25.7 727 -6.8 0 -2.5
TAGTGAGAGGAATTACTTTG
3551 SEQ ID NO: 1991 -18.9 -30.7 79 -97 -2.1 -7.5
AGAAGGAGCAGGAGGGAAAT
21 SEQ ID NO: 1992 -18.8 -38.5 88.9 -19.7 0 -2.7
GTCCGCAGTTCCCGCCGCAG
70 SEQ ID NO: 1993 -18.8 -45 97 -24.2 -2 -67
TAGAAGTCCATCACATCTCC
202 SEQ ID NO: 1994 -18.8 -35.9 84.3 -17.1 0 -3.5
GCTCCTCCTCTTAGGGTTTT
223 SEQ ID NO: 1995 -18.8 -37.3 88.5 -16.8 -1.7 -7.4
AATTACCCCAGGGGTGCAGA
989 SEQ ID NO: 1996 -18.8 -40.6 93 -14.9 -4.3 -21.9
CCAGTTTCTCTTGCTTAATT
1005 SEQ ID NO: 1997 -18.8 -30 78.4 -11.2 0 -2.8
GAACAGAAATGGCAGACATT
1077 SEQ ID NO: 1998 -18.8 -32.6 80.6 -12.3 -1.4 -5.1
GAATGACATTAAAAATAGGC
1176 SEQ ID NO: 1999 -18.8 -27.2 737 -8.4 0 -5
TATTCCAATTTTCGGAACCA
1209 SEQ ID NO:2000 -18.8 -30.7 78.7 -9.4 -2.5 -7.8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
CGACCAGGGAAGTTCAGAGT 1270 SEQ ID NO: 2001 -18.8 -38.8 89.2 -19.5 -0.1 -6
ATGACATCCTGAAGCTTCAT 1409 SEQ ID NO:2002 -18.8 -34.2 81.9 -11.8 0 -15.3
CTCCATAATGACATCCTGAA 1416 SEQ ID NO: 2003 -18.8 -33.3 81 -14.5 0 -4.6
AAGTTAAGACTCCATAATGA 1425 SEQ ID NO: 2004 -18.8 -29.7 77.1 -10.9 0 -4.9
TTGCCCATTTCACTGCTGCA 1848 SEQ ID NO: 2005 -18.8 -37.9 88.9 -19.1 0 -57
TGCCTTTGCCCATTTCACTG 1853 SEQ ID NO: 2006 -18.8 -36.6 87.4 -17.8 0 -2
ATACAGCATGTGTTTACATT 2057 SEQ ID NO: 2007 -18.8 -29.3 77 -9.6 -0.4 -9.3
TCTTGGCTCTTCAGACCGTC 2164 SEQ ID NO:2008 -18.8 -39.1 89 -18.4 -1.9 -9.4
AAACCGCTGCCAGTTCTGGC 2258 SEQ ID NO:2009 -18.8 -40.5 92.6 -16.6 -4.4 -18.2
ATAGAATCCAAGAGTTTTGT 2287 SEQ ID NO: 2010 -18.8 -29.3 76.3 -9 -1.4 -8
AACAAAACCTCTACAGGACA 2536 SEQ ID NO: 2011 -18.8 -33.6 83.7 -13.2 -1.5 -7.3
CTGGTGGTTTAGGTGCCATC 2761 SEQ ID NO: 2012 -18.8 -38.3 89.4 -13.3 -6.2 -14.8
CTTGCCATATAGCCATTGCA 3418 SEQ ID NO: 2013 -18.8 -35.1 85.2 -15.6 -0.5 -5.6
GTGTCTAGCCATTTTTGCCA 3600 SEQ ID NO: 2014 -18.8 -34.7 85.1 -15.9 0 -6.8
GTTTTGAGTTTACAGAAAGT 3855 SEQ ID NO: 2015 -18.8 -27.6 75.1 -7.6 -1.1 -4.3
CAAATTTCTTGTGGCTTAGT 3886 SEQ ID NO: 2016 -18.8 -29.8 78.4 -11 0 -3.8
CCTAGAGCAAACTGTTTGGT 4251 SEQ ID NO:2017 -18.8 -34.5 85.3 -14.1 -1.2 -10.6
GCTGACAGACTCACTGTTGG 4385 SEQ ID NO: 2018 -18.8 -38.4 88.4 -14 -5.6 -13.6
ATGTAAGCACCACCTTCCTG 4466 SEQ ID NO: 2019 -18.8 -37.1 88.1 -18.3 0 -3.2
ACTATTTACATATTGCTGGT 4592 SEQ ID NO: 2020 -18.8 -29.6 77.9 -10.8 0 -4.5
TGGACAGATCTGGCTGCTGC 351 SEQ ID NO: 2021 -18.7 -42.1 92.5 -19.3 -2.2 -16.4
GACCTATTGAGGTTTGCAAT 508 SEQ ID NO: 2022 -18.7 -32.7 81.6 -10.5 -3.5 -12.4
CTGAACTCTTGGGGTTCTCT 540 SEQ ID NO: 2023 -18.7 -37.2 87.4 -13 -4.9 -19
GGCAGCAGACACAGCAGTGG 563 SEQ ID NO: 2024 -18.7 -42.7 94.1 -22 -2 -9.1
CCCTTCAAATGTTGCTGTTC
628 SEQ ID NO:2025 -18.7 -33.3 82.6 -14.6 0 -1.8 GCCCTTCAAATGTTGCTGTT
629 SEQ ID NO:2026 -18.7 -34.3 84.4 -15.6 0 -1.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
TATCAACAGGTCTGATCTCC 770 SEQ ID NO:2027 -18.7 -35.9 84.3 -15.6 0 -11.1
CACCTATTCCAATTTTCGGA 1213 SEQ ID NO:2028 -18.7 -31.9 80.4 -12.1 -1 -4.5
ATTTTTTTCTTTGTTTTTCG 1603 SEQ ID NO: 2029 -18.7 -21.4 64.4 -2.7 0 0
AGTTGTGGTAACGTTGCAGG 1696 SEQ ID NO: 2030 -18.7 -35.9 87.4 -14 -1.9 -14.6
CAATCACTTGCCGCCCTCCT 1830 SEQ ID NO: 2031 -18.7 -40.8 92.1 -22.1 0 -3.9
TAGGTCATTCTAATTTCATC 2191 SEQ ID NO: 2032 -18.7 -28.9 75.2 -9.3 -0.7 -4
TCTTATCCAAAAATGTTTGG 2343 SEQ ID NO: 2033 -18.7 -27.7 74.6 -7 . -2 -9
TCAATACTCATGGTCTTATC 2356 SEQ ID NO:2034 -18.7 -31.4 78.5 -12.7 0 -5.2
AATTCAATACTCATGGTCTT 2359 SEQ ID NO: 2035 -18.7 -29.5 76.1 -10.8 0 -5.2
TTTGGTATCTGATTGGTGAT 2401 SEQ ID NO: 2036 -187 -31.9 79.5 -12.4 -0.6 -5.4
TTTTTTGATATTTCCATTTG 2426 SEQ ID NO: 2037 -187 -23 66.6 -4.3 0 -2.3
ACCATTCTTATTAAGGCAGT 2466 SEQ ID NO: 2038 -18.7 -31.6 80.8 -12.9 0 -7
ACAGTAATAGCTATAAAAGG 2960 SEQ ID NO: 2039 -18.7 -28.5 77.1 -8.9 0 -9.8
TAGAGTTTATTTGGGAAAGC 3092 SEQ ID NO: 2040 -18.7 -30.4 79.3 -11.7 0 -4.8
GATTGATTAATGTTTAGAGT 3106 SEQ ID NO:204i -18.7 -26.3 71.6 -7.6 0 -7.4
TCTTGGTAGTTATTTTTTAA 3369 SEQ ID NO: 2042 -18.7 -24.5 70.6 -5.8 0 -2.7
AAATAGGGCGTTAGGTACAG 3397 SEQ ID NO: 2043 -187 -34.3 85.7 -15.6 0 -7.2
TTTGCCATATTAGAGATTTA 3587 SEQ ID NO: 2044 -18.7 -27.5 74.1 -8.8 0 -2.5
ATTTTTGCCATATTAGAGAT 3590 SEQ ID NO: 2045 -187 -27.3 73.2 -7.7 -07 -3.8
ACACATTAGGGATGTGTAGT 3735 SEQ ID NO: 2046 -18.7 -34.2 84 -10.5 -5 -17.1
CGAGATAAACAACTTAGCTT 49 SEQ ID NO: 2047 -18.6 -29.6 77.9 -11 0 -3.8
AATCAACCAAAAGTCTTCGC 306 SEQ ID NO: 2048 -18.6 -31.5 79.9 -12.9 0 -3.2
CAGCAGTGGATGCTGAACTC
552 SEQ ID NO:2049 -18.6 -38.5 88.1 -13.6 -6.3 -15.8 ACAGCAGTGGATGCTGAACT
553 SEQ ID NO: 2050 -18.6 -38.3 88.6 -12.9 -6.8 -16.3 GCTGTTCTGAAGATACATCA
615 SEQ ID NO: 2051 -18.6 -33.3 81.1 -13.9 -0.5 -8.4
TTCTCCCGCCAGAGGAGAAA 806 SEQ ID NO: 2052 -18.6 -40.1 90.9 -15 -6.5 -15.8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CTCCATTATCCTTAATTTTG
897 SEQ ID NO:2053 -18.6 -27 73.4 -8.4 0 -07
ACATTTTATTACCAATTATA
1062 SEQ ID NO:2054 -18.6 -22.8 66.9 -4.2 0 -0.1
CATAGTGGTACATCTGTCCT
1116 SEQ ID NO:2055 -18.6 -35.6 85.2 -15.4 -1.5 -9.3
ATCCTGCTGTTGAGAAAGGG
1151 SEQ ID NO:2056 -18.6 -37.1 87.2 -16.9 -1.6 -6.8
TTGTTACCAGGATTTTCAGA
1666 SEQ ID NO:2057 -18.6 -31.5 80 -12.9 0 -4.3
TAGGGGTGAGTTGTGGTAAC
1704 SEQ ID NO: 2058 -18.6 -37.4 89.3 -18.8 0 -2.5
GTAGGGGTGAGTTGTGGTAA
1705 SEQ ID NO:2059 -18.6 -37.4 89.3 -18.8 0 -27
CACTTCAGGTTCAATAACCT
1742 SEQ ID NO:2060 -18.6 -32.5 817 -11.2 -2.7 -8.8
CACATGTAGTGCGTATTTAA
2592 SEQ ID NO:2061 -18.6 -29.8 78.5 -10 -0.6 -10.2
TATCCTAACTATACAGGGGG
2871 SEQ ID NO: 2062 -18.6 -35.9 87.2 -15.1 -2.2 -8.5
TGATTGATTAATGTTTAGAG
3107 SEQ ID NO: 2063 -18.6 -26.2 71.4 -7.6 0 -7.4
TTTTTTTTTGACAAGAATAC
3207 SEQ ID NO: 2064 -18.6 -22.8 67 -37 0 -7.8
CCTCTTGGTAGTTATTTTTT
3371 SEQ ID NO:2065 -18.6 -277 757 -8.5 -0.3 -2.8
GTCTAGCCATTTTTGCCATA
3598 SEQ ID NO:2066 -18.6 -32.8 82.4 -14.2 0 -3.2
TCTAAATCACAAAAATCAGT
3686 SEQ ID NO:2067 -18.6 -26.9 72.6 -8.3 0 0
ACACAAATAATTTGGCCACC
3918 SEQ ID NO:2068 -18.6 -32.2 81.8 -12.1 0 -11
TCACTGCACACAGGACCAGT
4288 SEQ ID NO:2069 -18.6 -40.6 91.8 -20.4 -1.6 -5.8
TCAGCAACCTTCACTGCACA
4298 SEQ ID NO:2070 -18.6 -38.1 88.9 -17.2 -2.3 -5.9
AAATAAACTCTTGTTGTAGG
4656 SEQ ID NO:2071 -18.6 -27.8 75.7 -9.2 0 -2.5
TCTGGCAGAGGAGCCGCTCG
99 SEQ ID NO:2072 -18.5 -45.1 96.3 -22.1 -4.5 -14.6
TTGAGAAGCGACAGCCAGTG
272 SEQ ID NO:2073 -18.5 -38.6 89.1 -19 -1 -7.1
TCACTTTTGTTTCTGTCTCT
405 SEQ ID NO: 2074 -18.5 -31.3 79.3 -12.8 0 0
CTCTTGGGGTTCTCTGGAAC
535 SEQ ID NO: 2075 -18.5 -38.4 88.8 -16.7 -3.1 -13.5
TGTGGTATACAATTTCACAT
671 SEQ ID NO:2076 -18.5 -29.5 76.9 -8.3 -27 -9
AGGAATGAATCGTCTTCTCC
820 SEQ ID NO:2077 -18.5 -34.8 82.6 -15.4 -0.7 -7.8
ATTATCCTTAATTTTGGGTT
893 SEQ ID NO: 2078 -18.5 -26.8 73.5 -5.9 -2.4 -10.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CATTATCCTTAATTTTGGGT
894 SEQ ID NO:2079 -18.5 -28 75.3 -7.2 -2.3 -10
TGACAAAACCAGATCTCCAT
911 SEQ ID NO: 2080 -18.5 -34 82.2 -15.5 0 -6.5
ATCTTCTTTTTCTGTTTTCA
959 SEQ ID NO: 2081 -18.5 -26.5 71.9 -8 0 0
GGATGCTGTATTCATGTCAT
1133 SEQ ID NO: 2082 -18.5 -33.4 80.8 -12.1 -0.8 -13.7
AATGACATTAAAAATAGGCT
1175 SEQ ID NO: 2083 -18.5 -26.9 73.4 -8.4 0 -3.7
CTGGATGGAGGAGAGCTTAC
1336 SEQ ID NO: 2084 -18.5 -39 89 -20 0 -7.6
CTTTACAGCTTCCACAAGTT
1440 SEQ ID NO:2085 -18.5 -32.1 82 -12.9 -0.4 -4
ATAGGTAATTGTGCTGTCCT
1482 SEQ ID NO:2086 -18.5 -34.1 84 -14.7 -0.8 -7.2
CATAGGTAATTGTGCTGTCC
1483 SEQ ID NO:2087 -18.5 -34.1 83.8 -14.7 -0.8 -6.6
CAGAGGTTTCTTGTGAGACT
1650 SEQ ID NO: 2088 -18.5 -34.9 84.1 -11.6 -4.8 -11.8
ATGCCATAAGAAACATCCAG
1929 SEQ ID NO: 2089 -18.5 -32.8 81.2 -14.3 0 -2.3
AGGTCATTCTAATTTCATCA
2190 SEQ ID NO:2090 -18.5 -29.7 76 -11.2 0 -3.4
AAACAAAACAACAGATGAAA
2573 SEQ ID NO:2091 -18.5 -26.1 71.9 -7.6 0 -2.5
AGTCTAGAAAATTTCATCCA
2834 SEQ ID NO: 2092 -18.5 -29.5 76.2 -10.5 0 -7.5
AATGCTATCCTAACTATACA
2876 SEQ ID NO: 2093 -18.5 -30.2 78.6 -11.7 0 -17
ATATACAAAATATGAGCTTT
3233 SEQ ID NO: 2094 -18.5 -25.1 70.2 -6.6 0 -6.4
AAAAATAGGGCGTTAGGTAC
3399 SEQ ID NO:2095 -18.5 -31.9 82.6 -13.4 0 -7.2
TCTGATTAAAAGTCTCTCAG
3531 SEQ ID NO:2096 -18.5 -30.8 78 -11.6 -0.4 -4.5
TGAGAGGAATTACTTTGTCT
3548 SEQ ID NO: 2097 -18.5 -31.8 79.9 -11.6 -1.7 -7.8
TTTACAGAAAGTTGGTAAGG
3847 SEQ ID NO: 2098 -18.5 -30 79.5 -10.9 -0.3 -57
TTGAGTTTACAGAAAGTTGG
3852 SEQ ID NO: 2099 -18.5 -29.9 78.5 -11.4 0 -4.6
CACAAATAATTTGGCCACCT
3917 SEQ ID NO:2100 -18.5 -32.1 81.8 -12.1 0 -11
ACCTATGAGATTCTGCACTA
4608 SEQ ID NO: 2101 -18.5 -34.6 83.7 -16.1 0 -5.7
AAACTCTTGTTGTAGGATAG
4652 SEQ ID NO: 2102 -18.5 -30.5 79.1 -11.5 -0.1 -6.5
ATAAACTCTTGTTGTAGGAT
4654 SEQ ID NO: 2103 -18.5 -29.5 77.2 -10.5 -0.1 -6.5
ATCACTTTTGTTTCTGTCTC
406 SEQ ID NO: 2104 -18.4 -30.3 77.5 -11.9 0 0 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecu Site oligo binding formation Duplex structure oligo oligo
AGACACAGCAGTGGATGCTG
557 SEQ ID NO: 2105 -18.4 -39.5 89.7 -15.9 -5.2 -14.1
TGTTGCTGTTCTGAAGATAC
619 SEQ ID NO: 2106 -18.4 -32.9 81.5 -14.5 0 -5.3
AGTTTCTCTTGCTTAATTAC
1003 SEQ ID NO: 2107 -18.4 -28.1 75.7 -97 0 -2.6
AAGGGATGCTGTATTCATGT
1136 SEQ ID NO:2108 -18.4 -34.1 82.9 -13.1 -0.4 -13.4
ATTCCAATTTTCGGAACCAA
1208 SEQ ID NO: 2109 -18.4 -30.3 78 -9.4 -2.5 -7.8
CCCAGAGAAGTCAAGTTGTC
1249 SEQ ID NO: 2110 -18.4 -36.1 85.6 -17.7 0 -3.9
TATTTTTTTCTTTGTTTTTC
1604 SEQ ID NO:2111 -18.4 -20.3 62.6 -1.9 0 0
ATCCTGCATATAACACTTCA
1755 SEQ ID NO: 2112 -18.4 -32.1 80.3 -13.7 0 -4.5
ACAGAGCTATCATATCCTGC
1768 SEQ ID NO:2113 -18.4 -36 84.8 -17.6 0 -5.2
CATGTTGAGCGTAGTCATGA
1808 SEQ ID NO:2114 -18.4 -34.6 83.2 -14.6 -1.4 -10.7
TGAAACCTGGTATTGCCTTT
1866 SEQ ID NO:2115 -18.4 -33.4 837 -13.4 -1.1 -10.7
GATGTAGGTCATTCTAATTT
2195 SEQ ID NO: 2116 -18.4 -28.7 75.3 -9.3 -0.9 -9
TTTTGGTATCTGATTGGTGA
2402 SEQ ID NO: 2117 -18.4 -31.7 79.7 -12.4 -0.8 -5.4
AACCTCTACAGGACAAACTG
2531 SEQ ID NO: 2118 -18.4 -34.8 85.2 -14.7 -1.7 -7.3
GCTTCTGTTGCCAAGTCTTG
2625 SEQ ID NO: 2119 -18.4 -35.6 85.8 -15.1 -0.3 -12.4
CTGCTTCTGTTGCCAAGTCT
2627 SEQ ID NO: 2120 -18.4 -36.8 87.4 -17.8 -0.3 -4.5
AAAAGTGATGACGACTCAAC
2646 SEQ ID NO:2121 -18.4 -31.9 79.6 -13.5 0 -5.5
CCATCTACTCTCCCATCACT
2667 SEQ ID NO: 2122 -18.4 -38 87.3 -19.6 0 0
ATCCAGCCAACTGTGAAAAA
2819 SEQ ID NO: 2123 -18.4 -33.3 82.7 -12.7 -2.2 -7.2
CTTGTAAACTTTTTTTCAGG
2915 SEQ ID NO: 2124 -18.4 -25.9 73.2 -7 -0.1 -3.5
TGATAGCTAAGTGCCATCAG
3157 SEQ ID NO: 2125 -18.4 -35.5 85.1 -12.5 -4.6 -11.2
GAAGTTATACAAACTACTTC
3473 SEQ ID NO:2126 -18.4 -27.4 74.8 -7.1 -1.9 -7.8
AGTAGCTGAGCTTTCCTGTA
3669 SEQ ID NO:2127 -18.4 -36.4 87.5 -16.2 0 -11.8
ACATTAGGGATGTGTAGTTT
3733 SEQ ID NO: 2128 -18.4 -317 80.8 -12.4 -0.8 -8.5
CATCGCTGCCTGTATGAATG
4222 SEQ ID NO: 2129 -18.4 -35.6 84.5 -16.5 -0.4 -57
TTCCTGTCTCCTGTTTACAT
4452 SEQ ID NO: 2130 -18.4 -34 83.3 -15.6 0 -4.4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CTTCCTGTCTCCTGTTTACA
4453 SEQ ID NO: 2131 -18.4 -35 85 -16.6 0 -2.6
CTGTTGTCATTCAACTGCTG
4536 SEQ ID NO:2132 -18.4 -33.4 82.2 -12.7 -2.3 -7.6
TTACATATTGCTGGTACCTC
4587 SEQ ID NO: 2133 -18.4 -33.3 83.1 -13.6 0 -10.6
AACTCTTGTTGTAGGATAGA
4651 SEQ ID NO: 2134 -18.4 -32 80.8 -13.1 -0.1 -7.9
CGCAGAAGGAGCAGGAGGGA
24 SEQ ID NO: 2135 -18.3 -43.5 947 -24.1 -1 -3.3
GAGATAAACAACTTAGCTTG
48 SEQ ID NO:2136 -18.3 -29.3 77.4 -11 0 -3.8
TAATGATTCTTTGGAGTCCA
134 SEQ ID NO: 2137 -18.3 -31.9 79.7 -9.8 -3.8 -14.2
ACTGAGCCTTTTGGAAAATC
322 SEQ ID NO:2138 -18.3 -32.3 81.1 -12.9 -1 -4.9
CATTGCTTACTGAGCCTTTT
330 SEQ ID NO:2139 -18.3 -32.2 81.8 -12.1 -1.4 -11.4
TTTCCCCCGAGGAAAGGCTG
462 SEQ ID NO: 2140 -18.3 -40.7 92.7 -19.2 -3.2 -11.3
TTGGTGCCAGTCTGGCCCTT
643 SEQ ID NO: 2141 -18.3 -42.5 95.2 -15.1 -6.9 -26.4
TCCCGCCAGAGGAGAAAGCA
803 SEQ ID NO: 2142 -18.3 -42.3 93.8 -21.7 -2.3 -7.3
CGAGTTTCCTTCCAAAAGGA
836 SEQ ID NO:2143 -18.3 -33.8 83.4 -12.5 -3 -9
TTATCCTTAATTTTGGGTTT
892 SEQ ID NO: 2144 -18.3 -26.6 73.6 -5.9 -2.4 -10.9
TTGACAAAACCAGATCTCCA
912 SEQ ID NO:2145 -18.3 -33.8 82.4 -15.5 0 -6.5
GCTGTATTCATGTCATAGTG
1129 SEQ ID NO:2146 -18.3 -32.2 80.1 -12.3 -1.3 -11
ACAGCTTCCACAAGTTAAGA
1436 SEQ ID NO: 2147 -18.3 -33.6 837 -14.1 -1.1 -57
TTTGAAGAAAACTTTACAGC
1451 SEQ ID NO:2148 -18.3 -27.5 75 -8.1 -1 -5.1
GCTGTCCTTCCACTGCTCTT
1470 SEQ ID NO:2149 -18.3 -39.5 90.3 -20 -1.1 -4.6
CCTGCTGAATTCCTTTTATT
1620 SEQ ID NO:2150 -18.3 -30.1 78.2 -11.8 0 -5.2
TGCAGGAACTATTGTTTTGT
1682 SEQ ID NO: 2151 -18.3 -31.1 80.1 -10.9 -1.9 -9
AACACTTCAGGTTCAATAAC
1744 SEQ ID NO: 2152 -18.3 -30.2 78.4 -11.9 0 -4.5
GTGTAAGTTCCTGAAACCTG
1877 SEQ ID NO: 2153 -18.3 -33.6 83.6 -15.3 0 -2.4
ACCACTTCATGCATAGAATC
2299 SEQ ID NO: 2154 -18.3 -33.2 81.1 -14 0 -9.8
CATTCTTATTAAGGCAGTCA
2464 SEQ ID NO: 2155 -18.3 -30.6 78.8 -12.3 0 -7.2
TTAAAACAAAACAACAGATG
2576 SEQ ID NO:2156 -18.3 -25 70.9 -6.7 0 -0.8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CTGAAAAGTGATGACGACTC
2649 SEQ ID NO:2157 -18.3 -33.3 81.1 -15 0 -5.5
GAAAATTTCATCCAGCCAAC
2828 SEQ ID NO:2158 -18.3 -31.3 79.2 -13 0 -6
TATACAAAATATGAGCTTTT
3232 SEQ ID NO:2159 -18.3 -24.9 70.2 -6.6 0 -6.4
ATATATACAAAATATGAGCT
3235 SEQ ID NO:2160 -18.3 -25.7 70.9 -7.4 0 -4.8
GGCCTCTTGGTAGTTATTTT
3373 SEQ ID NO:2161 -18.3 -32.6 82.9 -11.5 -2.8 -8.3
CAAACTACTTCAAAAGGTCC
3464 SEQ ID NO:2162 -18.3 -31.1 80.4 -12.8 0 -3.6
ATCACAAAAATCAGTAGCTG
3681 SEQ ID NO:2163 -18.3 -30.3 78 -11.5 0 -7.8
CACATTAGGGATGTGTAGTT
3734 SEQ ID NO:2164 -18.3 -32.9 82.2 -11.4 -3.2 -13.5
TCTCATCTAAAAATCATTCT
4116 SEQ ID NO: 2165 -18.3 -27.5 72.4 -9.2 0 0
TCTTTGACCCCTACAAAAAA
4161 SEQ ID NO:2166 -18.3 -31 80.5 -127 0 -2.4
TTCTTTGACCCCTACAAAAA
4162 SEQ ID NO:2167 -18.3 -31 80.5 -12.7 0 -2.4
TTTCTTTGACCCCTACAAAA
4163 SEQ ID NO:2168 -18.3 -31 80.5 -12.7 0 -2.4
CATCTCCCTCTTCCCCTAGA
4265 SEQ ID NO: 2169 -18.3 -40.3 90.6 -22 0 -4.3
TAACTTCCTCTGTAATCTCA
4331 SEQ ID NO:2170 -18.3 -32.2 80.6 -13.9 0 -0.3
TAGTGCAAGGGGAGATTGAG
4419 SEQ ID NO:2171 -18.3 -37.5 87.8 -19.2 0 -4.8
ACTTTACATACTTTAGTGCA
4432 SEQ ID NO:2172 -18.3 -29.3 78.2 -11 0 -6.3
ACATACTTTACATACTTTAG
4436 SEQ ID NO: 2173 -18.3 -26.2 73.4 -7.9 0 0
TGTTTACATACTTTACATAC
4441 SEQ ID NO: 2174 -18.3 -26.3 73.4 -8 0 -3.6
AGCTTCTGTTGTCATTCAAC
4541 SEQ ID NO: 2175 -18.3 -32.5 80.8 -13 -1.1 -5.2
ACCTCTATGCAAACTATTGC
4572 SEQ ID NO: 2176 -18.3 -32.9 82.3 -13.4 -1.1 -5
CTCATTTTATTTGGAAATAA
4670 SEQ ID NO:2177 -18.3 -23.4 67.5 -5.1 0 -4.1
GAAGCGACAGCCAGTGAGGG
268 SEQ ID NO: 2178 -18.2 -42.2 93.2 -22.9 -1 -7.1
TCAACCAAAAGTCTTCGCTG
304 SEQ ID NO: 2179 -18.2 -33.7 83.2 -15.5 0 -3.2
AGCCTTTTGGAAAATCAACC
318 SEQ ID NO: 2180 -18.2 -32 81.3 -127 -1 -4.6
GCCCTGCTGTGGGAATCCCA
437 SEQ ID NO:2181 -18.2 -44.1 95.8 -21.4 -4.5 -15.8
GCAGTGGATGCTGAACTCTT
550 SEQ ID NO: 2182 -18.2 -37.3 87 -16 -3.1 -11 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
AACAGTTTTCATCTATCAAC 783 SEQ ID NO: 2183 -18.2 -28.2 74.6 -10 0 -0.5
ATCTCCATTATCCTTAATTT 899 SEQ ID NO: 2184 -18.2 -27.5 73.2 -9.3 0 -0.7
TGGCACCTATTCCAATTTTC 1216 SEQ ID NO:2185 -18.2 -32.6 81.7 -13.6 -0.6 -4.3
CCTTGGCACCTATTCCAATT 1219 SEQ ID NO:2186 -18.2 -34.7 84.9 -15 -1.4 -6.5
CCAGAGAAGTCAAGTTGTCA 1248 SEQ ID NO: 2187 -18.2 -34.9 84.1 -16 -0.4 -3.7
CACACCAGGCAGAGTTTGGG 1384 SEQ ID NO:2188 -18.2 -40.1 91.8 -16.8 -5.1 -15.3
GATGCAATCATTCCTTCCAG 1505 SEQ ID NO:2189 -18.2 -34.1 81.8 -15.9 0 -6.3
CCACTTCATGCATAGAATCC 2298 SEQ ID NO: 2190 -18.2 -34.3 82.6 -15.1 0 -9.9
GATATTTCCATTTGAATATT 2420 SEQ ID NO: 2191 -18.2 -24.1 67.5 -4.3 -1.6 -6
GCCAAGTCTTGGCCCTCTAT 2616 SEQ ID NO: 2192 -18.2 -39.7 91 -147 -4.3 -21.8
ACTGCTTCTGTTGCCAAGTC 2628 SEQ ID NO: 2193 -18.2 -36.9 87.4 -18.1 -0.3 -4.5
TCACAAAAATCAGTAGCTGA 3680 SEQ ID NO:2194 -18.2 -31.6 79.8 -11.5 -0.7 -12
TCATTTTAATAAATTGCATG 4067 SEQ ID NO: 2195 -18.2 -23.4 67.3 -5.2 0 -4.6
GGATGTAAGCACCACCTTCC 4468 SEQ ID NO: 2196 -18.2 -38.6 89.4 -18.3 -2.1 -9.1
AACCTATGAGATTCTGCACT 4609 SEQ ID NO: 2197 -18.2 -34.2 83 -16 0 -6
AGATAAACAACTTAGCTTGT 47 SEQ ID NO: 2198 -18.1 -29.1 77.4 -11 0 -5.1
CGCCCGCCACCGTCCGCAGT 81 SEQ ID NO: 2199 -18.1 -47.5 100.4 -28.3 -1 -4.5
CCATCAGTGAATATCAACTC 117 SEQ ID NO:2200 -18.1 -32.2 79.3 -14.1 0 -6.7
AGCTCCTCCTCTTAGGGTTT 224 SEQ ID NO: 2201 -18.1 -38.5 90.1 -18.2 -2.2 -7.4
GTCTGGCCCTTCAAATGTTG 634 SEQ ID NO: 2202 -18.1 -35.7 857 -17.1 0 -7.5
GTTTAGTGTCCGGTAAAATG 876 SEQ ID NO: 2203 -18.1 -30.6 79.8 -12 0 -8
TGGATGGAGGAGAGCTTACA 1335 SEQ ID NO: 2204 -18.1 -39 89.3 -20.4 0 -7.6
ACAAGTTAAGACTCCATAAT 1427 SEQ ID NO: 2205 -18.1 -29.5 77.2 -11.4 0 -4.9
TGTTACCAGGATTTTCAGAG 1665 SEQ ID NO:2206 -18.1 -32.7 81.6 -14.6 0 -4.3
GGGTAGGGGTGAGTTGTGGT 1707 SEQ ID NO:2207 -18.1 -41.8 94.4 -23.7 0 -2.7
TTCAGGTTCAATAACCTCCA 1739 SEQ ID NO: 2208 -18.1 -33.9 83.4 -13 -2.8 -9.7 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CTTCAGGTTCAATAACCTCC
1740 SEQ ID NO: 2209 -18.1 -33.9 83.3 -13 -2.8 -9.7
TGAGTCTGGAACAGAGCTAT
1778 SEQ ID NO:2210 -18.1 -36.7 86.2 -16.4 -2 -12
GCAATCACTTGCCGCCCTCC
1831 SEQ ID NO:2211 -18.1 -42.1 93.3 -22.1 -1.9 -6.6
TGTCTATATGATCTCCACCC
1957 SEQ ID NO: 2212 -18.1 -36.2 84.9 -18.1 0 -4.2
ACTCTTCATAAGATACCTGA
2100 SEQ ID NO: 2213 -18.1 -32.4 80.4 -14.3 0 -4.3
TGAAGAGAGAAGCAGTAAGG
2135 SEQ ID NO: 2214 -18.1 -35.2 84.9 -17.1 0 -2.7
CATAGAATCCAAGAGTTTTG
2288 SEQ ID NO:2215 -18.1 -29.2 76.3 -11.1 0 -6.7
AATAGTTAAGGAGATTTTCA
2319 SEQ ID NO:2216 -18.1 -27.2 73.5 -8.2 0 -9.8
CGGGGAATTCAATACTCATG
2364 SEQ ID NO:2217 -18.1 -33.3 81.6 -13.6 -1.5 -7.1
TATACAGGGGGGGGATACAC
2862 SEQ ID NO: 2218 -18.1 -40.4 92.4 -22.3 0 0
GAGATTTGAGTCAATTTTTG
3185 SEQ ID NO:2219 -18.1 -26.7 72 -6.9 -0.7 -11.5
AGGACTGTTAATTTGCACAA
3280 SEQ ID NO: 2220 -18.1 -31.1 80.1 -13 0 -5.3
GTTATTTTTTAAATTAGTCT
3361 SEQ ID NO: 2221 -18.1 -21.1 64.5 -3 0 -1.9
AGTTATTTTTTAAATTAGTC
3362 SEQ ID NO: 2222 -18.1 -21.1 64.5 -3 0 -1.9
GCCATATAGCCATTGCAAAA
3415 SEQ ID NO: 2223 -18.1 -327 82.2 -13.9 -0.5 -6.7
ATACAAACTACTTCAAAAGG
3467 SEQ ID NO: 2224 -18.1 -27.8 75.7 -9.7 0 -1.9
GTTCTAAATCACAAAAATCA
3688 SEQ ID NO: 2225 -18.1 -25.7 70.9 -7.6 0 -0.4
TTTCTTGTGGCTTAGTAAAT
3882 SEQ ID NO: 2226 -18.1 -29 77.5 -10.9 0 -5
CTATTTTACAATCATTTTAA
4078 SEQ ID NO: 2227 -18.1 -21.2 64.1 -3.1 0 0
CAAGCTATTTTACAATCATT
4082 SEQ ID NO:2228 -18.1 -25.7 71.5 -7.6 0 -5
ATTTTACACTATACAAGCTA
4095 SEQ ID NO:2229 -18.1 -27.4 75.2 -9.3 0 -5
CATGATAAAACAATCTCATC
4129 SEQ ID NO: 2230 -18.1 -27.5 72.5 -9.4 0 -3.4
TGCTGGTACCTCTATGCAAA
4579 SEQ ID NO: 2231 -18.1 -36.3 87.4 -15.1 -3.1 -11.8
TAGTGTATTATTGGCAACCT
4624 SEQ ID NO: 2232 -18.1 -31.8 81.6 -13.7 0 -4.9
CAGAAGGAGCAGGAGGGAAA
22 SEQ ID NO:2233 -18 -39.5 90.3 -21.5 0 -27
TCGCCCGCCACCGTCCGCAG
82 SEQ ID NO:2234 -18 -47.7 100.2 -28.6 -1 -4.6 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TTGCTGTTCTGAAGATACAT
617 SEQ ID NO:2235 -18 -31.8 79.6 -13.3 -0.1 -8
CCTGTTGTTGCTGTTGAGGA
1357 SEQ ID NO:2236 -18 -36.8 87.4 -18.1 -0.4 -6.1
GTAGTGGCCTGCTGAATTCC
1627 SEQ ID NO:2237 -18 -38.6 89.3 -17.8 -2.2 -13.6
ATGATCCTCCAAGTTGAGTC
1792 SEQ ID NO: 2238 -18 -35.5 83.6 -16.6 -0.8 -8.2
TAAGAAACATCCAGGAGTAC
1923 SEQ ID NO:2239 -18 -33.2 82.5 -14.5 0 -8.7
TTAATAATCAGATCAGGAGC
2002 SEQ ID NO:2240 -18 -31.3 78.6 -13.3 0 -5.3
ACAGCATGTGTTTACATTGG
2055 SEQ ID NO: 2241 -18 -32.3 81.5 -13.4 -0.2 -9.4
AAGAGTTTTGTCAGTTGATA
2278 SEQ ID NO: 2242 -18 -29.2 76.4 -10.3 -07 -9.2
GAAGAGAAAGTTCATCACAC
2793 SEQ ID NO: 2243 -18 -31.8 79.1 -12.9 -07 -7
GTCTAGAAAATTTCATCCAG
2833 SEQ ID NO: 2244 -18 -29.5 76.4 -11 0 -7.5
GTAAACTTTTTTTCAGGTTT
2912 SEQ ID NO:2245 -18 -24.8 71.3 -5.6 -1.1 -4.6
GTAATAGCTATAAAAGGCAC
2957 SEQ ID NO: 2246 -18 -29.8 79 -9.3 -2.5 -9.8
TGCCCATCTTAAACAGCTGT
3061 SEQ ID NO: 2247 -18 -35.9 86.7 -16.1 0 -11.7
ATTTGGGAAAGCTACGAACT
3084 SEQ ID NO: 2248 -18 -32.8 82.5 -14.8 0 -5
AGCAGATATATACAAAATAT
3240 SEQ ID NO:2249 -18 -25.7 70.9 -7.7 0 -3.1
CTATATAATTCTCCACTGAA
3259 SEQ ID NO: 2250 -18 -29 75.9 -11 0 -3.4
TTCTAAATCACAAAAATCAG
3687 SEQ ID NO:2251 -18 -25.6 70.7 -7.6 0 0
AGGGATGTGTAGTTTTACAA
3728 SEQ ID NO:2252 -18 -31.5 81 -12.1 -1.3 -9
AGAAAGTTGGTAAGGTGCAC
3842 SEQ ID NO:2253 -18 -34.6 85.4 -16.1 0 -7.9
GTTTACAGAAAGTTGGTAAG
3848 SEQ ID NO:2254 -18 -28.9 77.8 -10.9 0 -4.7
AGTTTACAGAAAGTTGGTAA
3849 SEQ ID NO:2255 -18 -28.9 77.7 -10.9 0 -3.3
ATCAAATTTCTTGTGGCTTA
3888 SEQ ID NO: 2256 -18 -29 76.3 -11 0 -4
TATATTAGAAGTTCCATATT
3966 SEQ ID NO: 2257 -18 -25.7 71 -7.7 0 -4.9
AATATATTAGAAGTTCCATA
3968 SEQ ID NO:2258 -18 -257 71 -77 0 -4.9
TGATATATATCTGAAACTAT
4001 SEQ ID NO: 2259 -18 -26.2 70.8 -6.6 0 -11.4
ATGATATATATCTGAAACTA
4002 SEQ ID NO:2260 -18 -26.2 70.8 -6.6 0 -11.4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TATGATATATATCTGAAACT
4003 SEQ ID NO: 2261 -18 -26.2 70.8 -6.6 0 -11.4
CATTTTAATAAATTGCATGT
4066 SEQ ID NO: 2262 -18 -23.2 67.4 -5.2 0 -5.6
TTATTTTACACTATACAAGC
4097 SEQ ID NO: 2263 -18 -26.2 73.5 -8.2 0 -0.4
CGCTGACAGACTCACTGTTG
4386 SEQ ID NO: 2264 -18 -37.5 87.3 -15.9 -3.6 -9.3
CCGCAGTTCCCGCCGCAGCC
68 SEQ ID NO:2265 -17.9 -47.1 99.7 -27.2 -2 -7.5
GCCAGTGAGGGTGAAGACGC
259 SEQ ID NO: 2266 -17.9 -42.3 93.2 -22.4 -2 -8
TCTGATTGAGAAGCGACAGC
277 SEQ ID NO: 2267 -17.9 -36.9 85.7 -17.4 -1.5 -7.2
GACACAGCAGTGGATGCTGA
556 SEQ ID NO:2268 -17.9 -39.8 89.6 -15.1 -6.8 -16.3
TGCTGTTCTGAAGATACATC
616 SEQ ID NO: 2269 -17.9 -33.3 81.1 -14.9 -0.1 -8
ATTCGAGTTTCCTTCCAAAA
839 SEQ ID NO: 2270 -17.9 -30.4 78.1 -12.5 0 -6.3
GGTAAAATGAGAGGCTTGCA
865 SEQ ID NO:2271 -17.9 -34.9 85 -16.5 -0.1 -6.6
ATGAACAGAAATGGCAGACA
1079 SEQ ID NO: 2272 -17.9 -33.8 82.2 -14.9 -0.9 -5.1
TTTGTTTTTCGAGCTTCCAG
1594 SEQ ID NO: 2273 -17.9 -31.2 80 -13.3 0 -6.6
ATTGTTTTGTTACCAGGATT
1672 SEQ ID NO: 2274 -17.9 -28.8 76.5 -9.7 -1.1 -7.1
TGCATATAACACTTCAGGTT
1751 SEQ ID NO:2275 -17.9 -31.7 80.6 -13.8 0 -4.5
CATTTCACTGCTGCAATCAC
1843 SEQ ID NO: 2276 -17.9 -33.6 82.1 -15.2 0 -7.9
GGAACTGAAGAGAGAAGCAG
2140 SEQ ID NO: 2277 -17.9 -36.3 85.5 -18.4 0 -4.2
AGGAACTGAAGAGAGAAGCA
2141 SEQ ID NO: 2278 -17.9 -36.3 85.7 -18.4 0 -27
TTTTCCTAGCTCTTTGATGT
2210 SEQ ID NO: 2279 -17.9 -31.5 80 -13.6 0 -3.8
AAGCAATAGTTAAGGAGATT
2323 SEQ ID NO:2280 -17.9 -29.4 77.2 -11.5 0 -27
TCTACAGGACAAACTGATAG
2527 SEQ ID NO: 2281 -17.9 -33.2 82.2 -12.2 -3.1 -7.6
ATGTAGTGCGTATTTAAAAC
2589 SEQ ID NO: 2282 -17.9 -27.4 75.1 -8.6 0 -9.6
TATGAAGAGAAAGTTCATCA
2796 SEQ ID NO: 2283 -17.9 -29.8 76.1 -10.9 -0.9 -7
ATACAAAATATGAGCTTTTT
3231 SEQ ID NO: 2284 -17.9 -24.5 69.4 -6.6 0 -6.4
TATACAAACTACTTCAAAAG
3468 SEQ ID NO: 2285 -17.9 -25.8 72.6 -7.9 0 -0.1
AGTTATACAAACTACTTCAA
3471 SEQ ID NO: 2286 -17.9 -27.1 74.5 -8.6 -0.3 -2.8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TGTCTGATTAAAAGTCTCTC
3533 SEQ ID NO:2287 -17.9 -30.9 78.1 -13 0 -3.7
GTTAAGTTTTGAGTTTACAG
3860 SEQ ID NO:2288 -17.9 -26.5 74.1 -8.6 0 -4.2
TGCGCTGACAGACTCACTGT
4388 SEQ ID NO:2289 -17.9 -40 90.5 -20.2 -1.8 -11.1
CTATTTACATATTGCTGGTA
4591 SEQ ID NO:2290 -17.9 -28.7 76.8 -10.8 0 -6.5
AAATAATTTCTCCAAAATAC
4742 SEQ ID NO:2291 -17.9 -23.5 67.7 -5.6 0 0
GCCCGCCACCGTCCGCAGTT
80 SEQ ID NO: 2292 -17.8 -46 98.9 -27.3 -0.7 -4.6
AGTCCATCACATCTCCCCTC
198 SEQ ID NO: 2293 -17.8 -40.3 89.5 -22.5 0 -0.6
TGTGGGAATCCCAGGTCATT
430 SEQ ID NO: 2294 -17.8 -38.5 88.6 -15.9 -3.5 -177
GTGGTATACAATTTCACATT
670 SEQ ID NO: 2295 -17.8 -28.3 75.3 -8.3 -2.2 -8
AAATGAGAGGCTTGCAGTCC
861 SEQ ID NO:2296 -17.8 -37.2 87.1 -18 -1.1 -10.1
CCTTAATTTTGGGTTTAGTG
888 SEQ ID NO:2297 -17.8 -28.6 77.6 -97 -1 -7.4
CCCCAGGGGTGCAGAGTTCG
984 SEQ ID NO:2298 -17.8 -44.2 96.3 -22.6 -1.9 -15.8
AGATCCTTGGCACCTATTCC
1223 SEQ ID NO:2299 -17.8 -37.7 87.7 -19.9 0 -3.9
CTGTTGAGGAGCTGGATGGA
1347 SEQ ID NO:2300 -17.8 -39.8 89.8 -19.6 -2.4 -10.1
CACTGCTCTTTTGAAGAAAA
1460 SEQ ID NO:2301 -17.8 -29.7 77.6 -11.2 0 -9
ATAGCGGCATGCTGGGCAGT
1547 SEQ ID NO:2302 -17.8 -42.2 94.2 -19.5 -2.6 -17.9
GTTGCAGGAACTATTGTTTT
1684 SEQ ID NO:2303 -17.8 -29.9 78.5 -10.9 -1.1 -8.7
GTTGTGGTAACGTTGCAGGA
1695 SEQ ID NO: 2304 -17.8 -36.2 87.4 -15.2 -1.9 -14.6
TAACACTTCAGGTTCAATAA
1745 SEQ ID NO:2305 -17.8 -29.3 77.3 -11.5 0 -4.5
GGTTTTCATACAGAGATACT
2117 SEQ ID NO:2306 -17.8 -31 78.9 -11.6 -1.6 -4.8
TTTGATGTAGGTCATTCTAA
2198 SEQ ID NO:2307 -17.8 -29.7 77 -10.3 -0.9 -10.9
CTACAGGACAAACTGATAGT
2526 SEQ ID NO: 2308 -17.8 -33 82.4 -11.6 -3.6 -8.6
CTCTACAGGACAAACTGATA
2528 SEQ ID NO: 2309 -17.8 -33.2 82.2 -13.1 -2.3 -6
ACAAAACCTCTACAGGACAA
2535 SEQ ID NO: 2310 -17.8 -33.6 83.7 -14.2 -1.5 -7.3
AAATGCTATCCTAACTATAC
2877 SEQ ID NO:2311 -17.8 -29 76.9 -11.2 0 -17
AAAAATGCTATCCTAACTAT
2879 SEQ ID NO:2312 -17.8 -27.3 74.2 -9.5 0 -1.7 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GCTTTCCTGTACCATCAGGA
3660 SEQ ID NO: 2313 -17.8 -37.6 87.9 -14.1 -5.7 -13.8
GCTGAGCTTTCCTGTACCAT
3665 SEQ ID NO: 2314 -17.8 -37.4 87.9 -18.7 0 -9.7
TGTCTGACATACAGTTCTAA
3701 SEQ ID NO: 2315 -17.8 -32.1 80.6 -13.4 -0.4 -9.1
GCCCTTCCCTTCCCAGATTA
4034 SEQ ID NO:2316 -17.8 -39.8 91.2 -22 0 -1.1
CCATCGCTGCCTGTATGAAT
4223 SEQ ID NO:2317 -17.8 -36.8 86.2 -18.1 -0.7 -5.7
CAACCTTCACTGCACACAGG
4294 SEQ ID NO: 2318 -17.8 -37.8 887 -18.4 -1.5 -7.1
TCATTCAACTGCTGGGGGAG
4530 SEQ ID NO:2319 -17.8 -39.5 90 -20.8 -0.7 -9
CCATCACATCTCCCCTCTCC
195 SEQ ID NO:2320 -17.7 -41.4 90.7 -23.7 0 0
GTCTGTTTCCCCCGAGGAAA
467 SEQ ID NO:2321 -17.7 -38.7 89.6 -18.6 -2.4 -9.1
CAAAATGTCAAAGGTGCTTT
695 SEQ ID NO: 2322 -17.7 -29.4 77.6 -11.2 0 -7.6
GGAATGAATCGTCTTCTCCC
819 SEQ ID NO: 2323 -17.7 -36 84.1 -16.9 -1.3 -8.4
AGTAAACTGTGCCCAGTTTC
1017 SEQ ID NO: 2324 -17.7 -34.6 85.4 -13.2 -3.4 -15
GACTCCATAATGACATCCTG
1418 SEQ ID NO: 2325 -17.7 -34.6 82.5 -16.9 0 -3.2
TTTGTTACCAGGATTTTCAG
1667 SEQ ID NO: 2326 -17.7 -30 78.4 -12.3 0 -4.3
CTGAAACCTGGTATTGCCTT
1867 SEQ ID NO: 2327 -17.7 -34.6 85.1 -15.3 -1.1 -10.7
TACTCTTCATAAGATACCTG
2101 SEQ ID NO: 2328 -17.7 -31.3 79.5 -13.6 0 -4.3
AACCATTCTTATTAAGGCAG
2467 SEQ ID NO:2329 -17.7 -30.3 79 -12.6 0 -7.2
TCTTTAAGGCAACCATTCTT
2477 SEQ ID NO:2330 -177 -31.2 80.2 -13.5 0 -5.4
ATTAATTCGACTTTCTTTAA
2490 SEQ ID NO: 2331 -17.7 -23.9 68.1 -6.2 0 -4
CTATTAATTCGACTTTCTTT
2492 SEQ ID NO: 2332 -177 -25.1 70.3 -7.4 0 -4
ACAACAAAACCTCTACAGGA
2538 SEQ ID NO: 2333 -17.7 -33.6 83.7 -14.3 -1.5 -7.3
AAAACAAAACAACAGATGAA
2574 SEQ ID NO:2334 -17.7 -26.1 71.9 -8.4 0 -2.5
TGCTTCTGTTGCCAAGTCTT
2626 SEQ ID NO:2335 -17.7 -35.6 86.1 -17.3 -0.3 -4.5
AAGAGAAAGTTCATCACACA
2792 SEQ ID NO: 2336 -17.7 -31.5 79 -13.8 0 -7
TGAAGAGAAAGTTCATCACA
2794 SEQ ID NO:2337 -17.7 -31.7 78.9 -12.8 -1.1 -7.2
GGGGGATACACCAACAGAAA
2853 SEQ ID NO:2338 -17.7 -37.3 88.2 -17.5 -2.1 -7.1 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecu Site oligo binding formation Duplex structure oligo oligo
CAAAAATGCTATCCTAACTA
2880 SEQ ID NO:2339 -17.7 -28.3 76.2 -10.6 0 -1.7
AGCGTAGTTCACTAAATATA
3000 SEQ ID NO:2340 -17.7 -29.2 77.6 -11 -0.2 -5.6
TACAAAATATGAGCTTTTTT
3230 SEQ ID NO:2341 -17.7 -24.3 69.5 -6.6 0 -6.4
GAAGCAGATATATACAAAAT
3242 SEQ ID NO: 2342 -17.7 -26.6 72.3 -8.9 0 -3.5
AAACTACTTCAAAAGGTCCT
3463 SEQ ID NO: 2343 -17.7 -31.1 80.5 -13.4 0 -7
TTAGTGAGAGGAATTACTTT
3552 SEQ ID NO: 2344 -17.7 -29.5 77.3 -9.7 -2.1 -7.5
GACAGATGGGAATGTGAAAA
3623 SEQ ID NO: 2345 -17.7 -32.8 80.6 -14 -1 -4.3
ACTCTATGGCACACATTAGG
3745 SEQ ID NO:2346 -17.7 -35.3 85.4 -16.4 -1.1 -4.2
ATGATAAAACAATCTCATCT
4128 SEQ ID NO:2347 -17.7 -27.5 72.2 -9.8 0 -1.9
TAGGTTATCCATCAGCATTT
4180 SEQ ID NO: 2348 -177 -32 80.4 -13.1 -1.1 -5.2
TCATATAGGTTATCCATCAG
4185 SEQ ID NO: 2349 -17.7 -31.6 79.1 -12.7 -1.1 -5.2
CCACCTTCCTGTCTCCTGTT
4457 SEQ ID NO:2350 -17.7 -39.4 90.4 -217 0 0
GATGTAAGCACCACCTTCCT
4467 SEQ ID NO:2351 -17.7 -37.4 88 -19.7 0 -3.5
TTAGTGTATTATTGGCAACC
4625 SEQ ID NO:2352 -17.7 -30.6 79.9 -12.9 0 -4.9
TTTAAGTCTGTTTCCCCCGA
472 SEQ ID NO: 2353 -17.6 -35.2 86 -17.6 0 -2.4
ATACATCAGAGTGAGTTTTT
603 SEQ ID NO: 2354 -17.6 -29.4 76.3 -11.8 0 -3.5
AGCAAACAGTTTTCATCTAT
787 SEQ ID NO:2355 -17.6 -29.1 76.3 -11.5 0 -3.3
CCAGAGGAGAAAGCAAACAG
798 SEQ ID NO: 2356 -17.6 -35.7 857 -18.1 0 -2.7
CATTACTGGGGCTTGACAAA
924 SEQ ID NO: 2357 -17.6 -34.6 85.1 -17 0 -5.9
ACCAATTATATTTGCTCCAG
1052 SEQ ID NO:2358 -17.6 -30.5 78.8 -12.9 0 -4.6
ATGACATTAAAAATAGGCTT
1174 SEQ ID NO: 2359 -17.6 -26.9 73.4 -9.3 0 -4.1
CGGAACCAACGGGAATTGGT
1197 SEQ ID NO:2360 -17.6 -37.5 88.5 -16.9 -3 -8.4
TTCCAATTTTCGGAACCAAC
1207 SEQ ID NO:2361 -17.6 -31.4 80 -11.6 -2.2 -6.9
GAAAACTTTACAGCTTCCAC
1445 SEQ ID NO: 2362 -17.6 -31.2 80.4 -13.6 0 -4
CAGCACATAGGTAATTGTGC
1488 SEQ ID NO:2363 -17.6 -33.9 83.8 -10.8 -5.5 -12.9
TCAGGTTCAATAACCTCCAA
1738 SEQ ID NO: 2364 -17.6 -33.9 83.4 -13.5 -2.8 -9.7 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CATGCATAGAATCCAAGAGT
2292 SEQ ID NO:2365 -17.6 -33.1 81 -15.5 0 -7
GTTAAGGAGATTTTCAACCA
2315 SEQ ID NO:2366 -17.6 -30.3 78.4 -11.8 0 -9.6
GCTATTAATTCGACTTTCTT
2493 SEQ ID NO: 2367 -17.6 -27.6 74.2 -10 0 -4
CAACAAAACCTCTACAGGAC
2537 SEQ ID NO: 2368 -17.6 -33.6 83.6 -14.4 -1.5 -7.3
CTGTTGCCAAGTCTTGGCCC
2621 SEQ ID NO: 2369 -17.6 -40.1 91.9 -157 -4.3 -21.8
CTTCTGTTGCCAAGTCTTGG
2624 SEQ ID NO: 2370 -17.6 -35.5 85.8 -15.1 -0.3 -13.8
TCCAGCCAACTGTGAAAAAA
2818 SEQ ID NO:2371 -17.6 -33.1 83 -13.6 -1.9 -6.4
ATGATTGATTAATGTTTAGA
3108 SEQ ID NO:2372 -17.6 -25.2 69.2 -7.6 0 -7.4
TAGTTTTACAAACATGGATG
3719 SEQ ID NO: 2373 -17.6 -28 75.4 -10.4 0 -6.2
CATATAGGTTATCCATCAGC
4184 SEQ ID NO: 2374 -17.6 -32.6 80.8 -13.8 -1.1 -5.2
ATCTCCCCTCTCCTGAGCAA
188 SEQ ID NO:2375 -17.5 -41.2 91.4 -22.1 -1.6 -5.4
CATTTTATTACCAATTATAT
1061 SEQ ID NO:2376 -17.5 -21.7 64.9 -4.2 0 -07
GAGAAGTCAAGTTGTCATCT
1245 SEQ ID NO: 2377 -17.5 -33 80.6 -15 -0.2 -7.1
GATGGAGGAGAGCTTACATC
1333 SEQ ID NO: 2378 -17.5 -37.1 85.7 -16.7 -2.9 -10.2
CCATAATGACATCCTGAAGC
1414 SEQ ID NO:2379 -17.5 -34.3 82.6 -16.8 0 -4.6
CTTTGTTTTTCGAGCTTCCA
1595 SEQ ID NO:2380 -17.5 -31.2 80 -13.7 0 -6.6
TTGTTTTGTTACCAGGATTT
1671 SEQ ID NO: 2381 -17.5 -28.6 76.7 -9.9 -1.1 -7.1
CCGTCCTTAGGAACTGAAGA
2149 SEQ ID NO: 2382 -17.5 -36.7 87.1 -17.6 -0.7 -11
TGATGTAGGTCATTCTAATT
2196 SEQ ID NO:2383 -17.5 -29.9 76.8 -11.2 -0.7 -10
TTTTGTCAGTTGATAAAACC
2273 SEQ ID NO:2384 -17.5 -27.7 74.9 -6.3 -37 -15.2
TATTTCCATTTGAATATTTT
2418 SEQ ID NO:2385 -17.5 -22.4 65.1 -4.9 0 -4.1
CAGACTTTGGGCACTGGTGG
2774 SEQ ID NO:2386 -17.5 -40.1 91.8 -20.9 -0.1 -11.6
AGGGGGGGGATACACCAACA
2857 SEQ ID NO:2387 -17.5 -43 95.9 -23.5 -2 -7.1
GGGAAAGCTACGAACTAGCT
3080 SEQ ID NO:2388 -17.5 -36.7 88.1 -16.2 -3 -9.1
AACCTATATAATTCTCCACT
3262 SEQ ID NO:2389 -17.5 -30 77.8 -12.5 0 -3.4
TTAGGGATGTGTAGTTTTAC
3730 SEQ ID NO:2390 -17.5 -307 80.1 -13.2 0 -3.5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
AGAAATTCACAGGACTTGTG
3769 SEQ ID NO:2391 -17.5 -32.4 80.8 -12.2 -0.9 -13.5
TGAGTTTACAGAAAGTTGGT
3851 SEQ ID NO: 2392 -17.5 -31.2 80.3 -137 0 -3.2
CCCTCTTCCCCTAGAGCAAA
4260 SEQ ID NO: 2393 -17.5 -39.6 91.3 -19.4 -2.7 -7.8
AGGATAACTTCCTCTGTAAT
4335 SEQ ID NO: 2394 -17.5 -32.1 80.6 -11.7 -2.9 -8.2
AGGATGTAAGCACCACCTTC
4469 SEQ ID NO:2395 -17.5 -37.4 88 -17.6 -2.3 -9.1
CAACTGCTGGGGGAGGGCTG
4525 SEQ ID NO:2396 -17.5 -44.8 97.2 -26.1 -1.1 -9
TCAACTGCTGGGGGAGGGCT
4526 SEQ ID NO:2397 -17.5 -45.1 97.8 -26.3 -1.1 -9.4
TATTTACATATTGCTGGTAC
4590 SEQ ID NO:2398 -17.5 -28.8 76.9 -10.8 0 -8
ACATGTCCTCATTTTATTTG
4677 SEQ ID NO:2399 -17.5 -28 74.4 -10.5 0 -4.8
AGAAGTCCATCACATCTCCC
201 SEQ ID NO:2400 -17.4 -37.9 86.6 -20.5 0 -2.4
CCCATATACAGTCCCATTGA
385 SEQ ID NO: 2401 -17.4 -35.6 85.1 -18.2 0 -3.4
ACACAGCAGTGGATGCTGAA
555 SEQ ID NO: 2402 -17.4 -38.3 88.6 -14.1 -6.8 -16.3
TCTCTGTGGGGGCAGCAGAC
573 SEQ ID NO: 2403 -17.4 -44.3 95.6 -22.4 -4.5 -12.8
AAGCAAACAGTTTTCATCTA
788 SEQ ID NO:2404 -17.4 -28.9 76.5 -11.5 0 -3.3
GGAATGACATTAAAAATAGG
1177 SEQ ID NO: 2405 -17.4 -27.1 73.6 -9.7 0 -5
GCTGGATGGAGGAGAGCTTA
1337 SEQ ID NO: 2406 -17.4 -40.2 90.6 -21.9 -0.8 -7.6
CGAGCTTCCAGGTTCATTCC
1585 SEQ ID NO: 2407 -17.4 -37.7 87.4 -15.7 -4.6 -13
AGAGGTTTCTTGTGAGACTC
1649 SEQ ID NO:2408 -17.4 -35.2 84.1 -12.9 -4.9 -12.8
GTAGGGTCATTTGGTCATCC
1899 SEQ ID NO:2409 -17.4 -36.6 86.1 -17.2 -2 -10.8
TTCATGCATAGAATCCAAGA
2294 SEQ ID NO:2410 -17.4 -32.1 79.3 -14 0 -9.1
CAACCACTTCATGCATAGAA
2301 SEQ ID NO:2411 -17.4 -32.7 81.3 -14.8 0 -8.1
TTAATTCGACTTTCTTTAAG
2489 SEQ ID NO:2412 -17.4 -24.9 70.4 -7.5 0 -3.8
TATTAATTCGACTTTCTTTA
2491 SEQ ID NO:2413 -17.4 -24.3 69 -6.9 0 -4
ACTGGTGGTTTAGGTGCCAT
2762 SEQ ID NO: 2414 -17.4 -38.1 90 -16.1 -4.6 -12.3
ATGAAGAGAAAGTTCATCAC
2795 SEQ ID NO:2415 -17.4 -30.7 77.2 -11.8 -1.4 -8.1
CAGGGGGGGGATACACCAAC
2858 SEQ ID NO:2416 -17.4 -43 95.3 -23.6 -2 -7.1 ι _
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
ACAGGGGGGGGATACACCAA 2859 SEQ ID NO: 2417 -17.4 -43 95.9 -23.6 -2 -7.1
ATACAGGGGGGGGATACACC 2861 SEQ ID NO: 2418 -17.4 -42.4 94.6 -24.4 -0.3 -4
TGTACAATAACTTGAATAAA 3044 SEQ ID NO: 2419 -17.4 -24.8 70.4 -7.4 0 -6.4
TGATTAATGTTTAGAGTTTA 3103 SEQ ID NO: 2420 -17.4 -25 70.4 -7.6 0 -3
ACTGTTAATTTGCACAACCT 3277 SEQ ID NO: 2421 -17.4 -30.9 80.3 -13 -0.1 -5.4
TTTTTTAAATTAGTCTTTTG 3357 SEQ ID NO: 2422 -17.4 -20.4 63.3 -3 0 -1.9
TTTTGCCATATTAGAGATTT 3588 SEQ ID NO: 2423 -17.4 -27.1 73.3 -9.7 0 -0.9
CCATCAGGAAAGATTAACCA 3649 SEQ ID NO: 2424 -17.4 -32.8 81.4 -14.8 -0.3 -3.6
ATTTCTTTGACCCCTACAAA 4164 SEQ ID NO: 2425 -17.4 -31.2 80.3 -13.8 0 -1
CACTGCACACAGGACCAGTC 4287 SEQ ID NO:2426 -17.4 -40.6 91.3 -21.6 -1.5 -10
TAACATTCAAAAAGCATTCA 4709 SEQ ID NO: 2427 -17.4 -26.6 72.5 -9.2 0 -2.7
CTCCAAAATACTGAAAATAA 4733 SEQ ID NO: 2428 -17.4 -25.7 71.6 -8.3 0 -0.4
CAGTAGCTCCTCCTCTTAGG 228 SEQ ID NO: 2429 -17.3 -38.9 90.1 -21 -0.1 -8.2
GAGCCTTTTGGAAAATCAAC 319 SEQ ID NO: 2430 -17.3 -31.1 79.4 -12.7 -1 -6.2
ATTGAGAGTGAAACTGCTTT 370 SEQ ID NO:243l -17.3 -31.2 79.2 -12.2 -1.7 -7.6
CGGTAAAATGAGAGGCTTGC 866 SEQ ID NO: 2432 -17.3 -35.2 85.4 -17.9 0 -6.5
GATCTCCATTATCCTTAATT 900 SEQ ID NO: 2433 -17.3 -29 75.1 -11.7 0 -4.2
AGAGAAGTCAAGTTGTCATC 1246 SEQ ID NO: 2434 -17.3 -33 80.6 -15 -0.4 -3.8
CTTCCACTGCTCTTTTGAAG 1464 SEQ ID NO: 2435 -17.3 -33.3 82.6 -16 0 -4.5
TGCTGTCCTTCCACTGCTCT 1471 SEQ ID NO: 2436 -17.3 -40.7 91.7 -22.2 -1.1 -4.6
TTGCAGGAACTATTGTTTTG 1683 SEQ ID NO:2437 -17.3 -29.8 78.4 -10.9 -1.5 -7.6
GAACAGAGCTATCATATCCT 1770 SEQ ID NO: 2438 -17.3 -33.8 81.8 -16.5 0 -5.2
TCCAAAAATGTTTGGAAGCA 2338 SEQ ID NO: 2439 -17.3 -30.8 79.3 -8.9 -4.6 -14.2
AAACCTCTACAGGACAAACT 2532 SEQ ID NO: 2440 -17.3 -33.6 83.8 -147 -1.5 -7.3
TCTGTTGCCAAGTCTTGGCC 2622 SEQ ID NO: 2441 -17.3 -39.2 90.5 -15.1 -4.3 -21.8
AAGTCTAGAAAATTTCATCC 2835 SEQ ID NO: 2442 -17.3 -28.3 74.7 -10.5 0 -7.5 kcal/mol kcal/mol degC kcal/mol kcal/mol kcal/mol Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
ATCAAAAATGCTATCCTAAC
2882 SEQ ID NO: 2443 -17.3 -28.4 75.3 -11.1 0 -1.7 TGTTAAAACCAGACAGTAAT
2972 SEQ ID NO: 2444 -17.3 -29.1 77.4 -11.1 -0.5 -3.4 ATAACTTGAATAAAAAATTA
3038 SEQ ID NO: 2445 -17.3 -20 61.6 -2.7 0 -0.3 TTAGAGTTTATTTGGGAAAG
3093 SEQ ID NO: 2446 -17.3 -27.9 75.6 -10.6 0 -3.8 TTTTTTTTTTGACAAGAATA
3208 SEQ ID NO: 2447 -17.3 -21.5 64.4 -3.7 0 -7.6 GCCTCTTGGTAGTTATTTTT
3372 SEQ ID NO: 2448 -17.3 -30.2 79.4 -10.5 -2.4 -7 CTGATTAAAAGTCTCTCAGC
3530 SEQ ID NO: 2449 -17.3 -31.8 79.8 -13.6 -0.7 -5.1 CCATTTTTGCCATATTAGAG
3592 SEQ ID NO:2450 -17.3 -29.2 77 -11.9 0 -0.9 AAGATTAACCAATTGGTGAC
3640 SEQ ID NO: 2451 -17.3 -30.3 78.3 -9.3 -2.2 -15.6 ACAAAAATCAGTAGCTGAGC
3678 SEQ ID NO:2452 -17.3 -32.6 81.7 -12.5 -1.5 -13.8 TCTATGGCACACATTAGGGA
3743 SEQ ID NO: 2453 -17.3 -367 87 -17.9 -1.4 -4.8 ATATATATCTGAAACTATAA
3999 SEQ ID NO:2454 -17.3 -23.9 67.7 -6.6 0 -3.4 GAAAGGAATTAGTGTATTAT
4633 SEQ ID NO: 2455 -17.3 -26.4 72.5 -9.1 0 -4.3 CCCGCCACCGTCCGCAGTTC
79 SEQ ID NO: 2456 -17.2 -45 97.1 -26.9 -07 -4.6 TGAGCCTTTTGGAAAATCAA
320 SEQ ID NO: 2457 -17.2 -31 79.3 -12.7 -1 -7.6 TACTGAGCCTTTTGGAAAAT
323 SEQ ID NO: 2458 -17.2 -31.2 80.2 -12.9 -1 -4.9 GGTCATTTCCCATCACTTTT
417 SEQ ID NO: 2459 -17.2 -32.5 80.8 -14.8 -0.2 -3.4 CCTTCAAATGTTGCTGTTCT
627 SEQ ID NO: 2460 -17.2 -32.1 80.9 -14.9 0 -1.8 CTTGACAAAACCAGATCTCC
913 SEQ ID NO: 2461 -17.2 -33.8 82.4 -16.6 0 -6.6 TTAATTACCCCAGGGGTGCA
991 SEQ ID NO: 2462 -17.2 -38.3 91 -14.3 -4.1 -21.7 GCTTGAATAGCCATTAGAAA
1298 SEQ ID NO: 2463 -17.2 -30.6 79 -11.8 -1.5 -6 TACAGCTTCCACAAGTTAAG
1437 SEQ ID NO: 2464 -17.2 -32.5 82.8 -14.1 -1.1 -57 GGTTCATTCCAGCCTGAAGA
1575 SEQ ID NO: 2465 -17.2 -37.4 87.1 -17.9 -2.3 -7 CCATAAGAAACATCCAGGAG
1926 SEQ ID NO: 2466 -17.2 -34 82.9 -16.3 0 -7.6 CAGAGCAAATGCCATAAGAA
1937 SEQ ID NO: 2467 -17.2 -32.6 81.4 -14.6 -0.6 -6.1 ATGCAGGGTAGAGTCATTCT
2029 SEQ ID NO:2468 -17.2 -36.6 86.1 -18.5 -0.8 -8.3
I57 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
AACATACAGCATGTGTTTAC 2060 SEQ ID NO: 2469 -17.2 -30.4 79 -11.5 -1.7 -9.8
CCTTAGGAACTGAAGAGAGA 2145 SEQ ID NO: 2470 -17.2 -35.4 84.9 -18.2 0 -7.1
GGAATTCAATACTCATGGTC 2361 SEQ ID NO: 2471 -17.2 -32.2 79.5 -15 0 -6.7
AACAACAAAACCTCTACAGG 2539 SEQ ID NO: 2472 -17.2 -32.1 82.1 -14.2 -0.4 -5.6
TCAAAAATGCTATCCTAACT 2881 SEQ ID NO: 2473 -17.2 -29.4 77.2 -12.2 0 -17
ACTTGTAAACTTTTTTTCAG 2916 SEQ ID NO: 2474 -17.2 -24.8 71.1 -7.6 0 -1.1
GAATAAAAAATTATGTAAGA 3031 SEQ ID NO: 2475 -17.2 -21.5 64.4 -4.3 0 -4.2
TGGGAAAGCTACGAACTAGC 3081 SEQ ID NO: 2476 -17.2 -36.7 88.1 -17.9 -1.6 -6
ATTGATTAATGTTTAGAGTT 3105 SEQ ID NO: 2477 -17.2 -24.8 69.4 -7.6 0 -6
TTTTTTTTTTTTGACAAGAA
3210 SEQ ID NO: 2478 -17.2 -20.9 63.4 -37 0 -6.1 TTTTTTTTTTTTTGACAAGA
3211 SEQ ID NO:2479 -17.2 -20.9 63.4 -3.7 0 -4.3 TTAATTTGCACAACCTATAT
3273 SEQ ID NO: 2480 -17.2 -27.1 74.2 -9.9 0 -4.4
ACAAACTACTTCAAAAGGTC 3465 SEQ ID NO: 2481 -17.2 -30 78.7 -12.8 0 -2.3
CTGTACCATCAGGAAAGATT 3654 SEQ ID NO: 2482 -17.2 -32.9 81.4 -14.1 -1.6 -5.6
AGTTTTGAGTTTACAGAAAG 3856 SEQ ID NO:2483 -17.2 -27.5 74.9 -9.1 -1.1 -4.3
ATTTCTTGTGGCTTAGTAAA 3883 SEQ ID NO: 2484 -17.2 -29 77.5 -11.8 0 -5
GATATATATCTGAAACTATA 4000 SEQ ID NO: 2485 -17.2 -25.4 70 -6.6 0 -11.4
GAATACCAATATGATATATA 4012 SEQ ID NO: 2486 -17.2 -25.3 69.8 -8.1 0 -4.8
AGCCCTTCCCTTCCCAGATT 4035 SEQ ID NO: 2487 -17.2 -40.6 92.1 -23.4 0 -1.3
ATCATTTTAATAAATTGCAT 4068 SEQ ID NO: 2488 -17.2 -22.4 65.1 -5.2 0 -4.6
GCGCTGACAGACTCACTGTT 4387 SEQ ID NO: 2489 -17.2 -38.8 89 -18.7 -2.7 -13.1
CCTGCGCTGACAGACTCACT 4390 SEQ ID NO:2490 -17.2 -41.1 91.8 -22.5 -1.3 -7.5
ACTTTAGTGCAAGGGGAGAT 4423 SEQ ID NO: 2491 -17.2 -36.1 867 -17.4 -0.1 -11
ATACTTTAGTGCAAGGGGAG 4425 SEQ ID NO: 2492 -17.2 -35 85.9 -17 -0.1 -9.3
TAAGGATGTAAGCACCACCT 4471 SEQ ID NO: 2493 -17.2 -36.3 87.5 -16.8 -2.3 -9.1
ACTTCTGAAGCTTCTGTTGT 4549 SEQ ID NO: 2494 -17.2 -33.5 82.7 -14 0 -12.7 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
CTTAATTTTGGGTTTAGTGT
887 SEQ ID NO: 2495 -17.1 -27.5 75.8 -10.4 0 -5.5
TATCCTTAATTTTGGGTTTA
891 SEQ ID NO: 2496 -17.1 -27 74.4 -7.5 -2.4 -10.9
CAGTGTTACATTACTGGGGC
932 SEQ ID NO: 2497 -17.1 -36.1 87.3 -14.6 -4.4 -10.6
AGCTGGATGGAGGAGAGCTT
1338 SEQ ID NO:2498 -17.1 -41 91.5 -22.5 -1.3 -57
CCACTGCTCTTTTGAAGAAA
1461 SEQ ID NO: 2499 -17.1 -32.1 81.1 -14.3 0 -9
CATACAGCATGTGTTTACAT
2058 SEQ ID NO:2500 -17.1 -30.5 78.6 -12.3 -0.7 -9.8
TAGGAACTGAAGAGAGAAGC
2142 SEQ ID NO:2501 -17.1 -35.5 84.8 -18.4 0 -2.9
TCATGCATAGAATCCAAGAG
2293 SEQ ID NO: 2502 -17.1 -33.3 80.9 -15.7 0 -7.7
GACTTTCTTTAAGGCAACCA
2482 SEQ ID NO: 2503 -17.1 -32.3 82.1 -15.2 0 -5.4
AAACAACAAAACCTCTACAG
2540 SEQ ID NO: 2504 -17.1 -29.7 78.6 -12.6 0 0
TTTAAAACAAAACAACAGAT
2577 SEQ ID NO: 2505 -17.1 -23.8 68.8 -6.7 0 -1.4
ATTTAAAACAAAACAACAGA
2578 SEQ ID NO: 2506 -17.1 -23.8 68.8 -6.7 0 -2.2
GTGCGTATTTAAAACAAAAC
2584 SEQ ID NO: 2507 -17.1 -25.6 72.8 -8.5 0 -7.3
TGTAGTGCGTATTTAAAACA
2588 SEQ ID NO:2508 -17.1 -28.4 77 -10.4 0 -9.6
TCTAATTTCTGGGATATATT
2702 SEQ ID NO:2509 -17.1 -27.9 74 -10.8 0 -6.8
ATTACGTCCACATATTAAGG
2724 SEQ ID NO:2510 -17.1 -30.1 78.4 -13 0 -3
TACAGGGGGGGGATACACCA
2860 SEQ ID NO:2511 -17.1 -43.4 96.5 -24.3 -2 -7.1
AAAATGCTATCCTAACTATA
2878 SEQ ID NO: 2512 -17.1 -27.7 75 -10.6 0 -17
AATGTTTAGAGTTTATTTGG
3098 SEQ ID NO: 2513 -17.1 -25.5 71.7 -8.4 0 -3.8
TAAGAAGTTATACAAACTAC
3476 SEQ ID NO: 2514 -17.1 -26.3 73.6 -8.6 -0.3 -3.6
GTTACAGCTGGTTATCTGGA
3507 SEQ ID NO: 2515 -17.1 -35.4 85.6 -16.6 -1.4 -11.1
TGTTACAGCTGGTTATCTGG
3508 SEQ ID NO: 2516 -17.1 -35.1 85.7 -16.6 -0.4 -10.6
ATTAGGGATGTGTAGTTTTA
3731 SEQ ID NO: 2517 -17.1 -29.6 78.1 -12.5 0 -3.1
TGTTAAACTCTATGGCACAC
3751 SEQ ID NO: 2518 -17.1 -32.8 82.5 -15 -0.5 -4.2
TTAAGTTTTGAGTTTACAGA
3859 SEQ ID NO: 2519 -17.1 -26.7 73.8 -9.6 0 -2.6
ACCTTGAATAGAAATCAAAT
3901 SEQ ID NO: 2520 -17.1 -27 72.5 -9.4 -0.2 -3.8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
TATATATCTGAAACTATAAC 3998 SEQ ID NO: 2521 -17.1 -25 70.1 -7.9 0 -2.2
AGATGGTGCCTTTAAGGATG 4483 SEQ ID NO: 2522 -17.1 -35 84.7 -16.8 -0.7 -9.8
GGGCTGTATGTGAAAGTAAC 4511 SEQ ID NO: 2523 -17.1 -34 84.2 -16.4 -0.2 -4.9
AGGAATTAGTGTATTATTGG 4630 SEQ ID NO: 2524 -17.1 -28.5 76.1 -11.4 0 -4.3
AAGGAGCAGGAGGGAAATAT
19 SEQ ID NO: 2525 -17 -36.4 86.4 -19.4 0 -2.5 GAAGGAGCAGGAGGGAAATA
20 SEQ ID NO: 2526 -17 -37.7 87.9 -207 0 -2.7 GGCAGAGGAGCCGCTCGCCC
96 SEQ ID NO: 2527 -17 -48.5 100.1 -24.9 -6.6 -19.8
TTACTGAGCCTTTTGGAAAA 324 SEQ ID NO: 2528 -17 -31 80.5 -12.9 -1 -4.9
GTCTCTCCCATATACAGTCC 391 SEQ ID NO: 2529 -17 -37.3 86.5 -20.3 0 -0.7
CTATCAACAGGTCTGATCTC 771 SEQ ID NO:2530 -17 -34.7 82.6 -16.1 0 -11.1
CTCACACCATGAACAGAAAT 1087 SEQ ID NO: 2531 -17 -32.6 80.5 -15.6 0 -4.1
ATAGGCTTCTGATCCTGCTG 1162 SEQ ID NO: 2532 -17 -37.7 87.4 -19.1 -1.6 -9.8
CAGAGAAGTCAAGTTGTCAT 1247 SEQ ID NO: 2533 -17 -32.7 80.6 -15 -0.4 -3.8
ACACCAGGCAGAGTTTGGGA 1383 SEQ ID NO:2534 -17 -40.4 92.2 -18.3 -5.1 -15.3
AATTGTGCTGTCCTTCCACT 1476 SEQ ID NO: 2535 -17 -35.8 85.8 -18.8 0 -3.8
CTATTGTTTTGTTACCAGGA 1674 SEQ ID NO: 2536 -17 -30.2 79.2 -13.2 0 -4.1
AATCACTTGCCGCCCTCCTA 1829 SEQ ID NO:2537 -17 -40 91.7 -23 0 -3.9
CAGGTGTAAGTTCCTGAAAC 1880 SEQ ID NO:2538 -17 -33.6 83.6 -12.1 -4.5 -11.5
AGGGTCATTTGGTCATCCAG 1897 SEQ ID NO:2539 -17 -37.3 86.9 -17.7 -2.6 -10.8
GAGTACTGCAGTAGGGTCAT 1909 SEQ ID NO: 2540 -17 -37.9 88.5 -17 -0.1 -16
GATACTCTTCATAAGATACC
2103 SEQ ID NO: 2541 -17 -30.6 77.8 -13.6 0 -4.2 AGATACTCTTCATAAGATAC
2104 SEQ ID NO: 2542 -17 -29.4 75.9 -12.4 0 -4.3 GAACTGAAGAGAGAAGCAGT
2139 SEQ ID NO: 2543 -17 -35.2 84 -17.1 -1 -7.2
AGCTCTTGGCTCTTCAGACC 2167 SEQ ID NO: 2544 -17 -39.7 90.1 -21.3 -1.3 -6.9
TTTCCTAGCTCTTTGATGTA 2209 SEQ ID NO: 2545 -17 -31.9 80.7 -14.9 0 -3.8
AATATTTTGGTATCTGATTG 2406 SEQ ID NO: 2546 -17 -26.1 71.3 -9.1 0 -5.4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
ATATTTCCATTTGAATATTT
2419 SEQ ID NO:2547 -17 -22.6 65.1 -5.6 0 -4.1
CCATTCTTATTAAGGCAGTC
2465 SEQ ID NO: 2548 -17 -31.8 80.6 -14.8 0 -7.2
TGCCAAGTCTTGGCCCTCTA
2617 SEQ ID NO:2549 -17 -407 92.9 -16.9 -4.3 -21.8
CAACTGCTTCTGTTGCCAAG
2630 SEQ ID NO:2550 -17 -35.3 85.8 -16.6 -1.7 -5.6
TCAACTGCTTCTGTTGCCAA
2631 SEQ ID NO:2551 -17 -35.6 86 -16.8 -1.8 -5.8
GTATGAAGAGAAAGTTCATC
2797 SEQ ID NO: 2552 -17 -29.9 76.4 -12.1 -0.6 -5.6
GCTGCCCATCTTAAACAGCT
3063 SEQ ID NO:2553 -17 -37.1 88.1 -16.8 -3.3 -10.1
TTGATTAATGTTTAGAGTTT
3104 SEQ ID NO: 2554 -17 -24.6 69.5 -7.6 0 -3.6
CACAAAAATCAGTAGCTGAG
3679 SEQ ID NO:2555 -17 -31.3 79.8 -11.5 -1.5 -13.8
CATTAGGGATGTGTAGTTTT
3732 SEQ ID NO:2556 -17 -30.4 79 -13.4 0 -4.9
ACAAGCTATTTTACAATCAT
4083 SEQ ID NO:2557 -17 -27 73.3 -10 0 -5
TTTTACACTATACAAGCTAT
4094 SEQ ID NO:2558 -17 -27.4 75.2 -10.4 0 -5
AACCTTCACTGCACACAGGA
4293 SEQ ID NO:2559 -17 -38.1 89.1 -19.5 -1.5 -8.8
AACATTCAAAAAGCATTCAA
4708 SEQ ID NO:2560 -17 -26.2 71.7 -9.2 0 -27
AGTTAATGATTCTTTGGAGT
137 SEQ ID NO:2561 -16.9 -29.3 76.3 -11.7 -0.5 -7.6
GCAAGCACACTGCTGGGGTT
172 SEQ ID NO: 2562 -16.9 -41.2 93.5 -22.1 -2.2 -9.3
CAACAGGTCTGATCTCCAAG
767 SEQ ID NO:2563 -16.9 -36.2 85.2 -177 -0.9 -11.1
TCCGGTAAAATGAGAGGCTT
868 SEQ ID NO:2564 -16.9 -35.4 85.6 -18.5 0 -6.3
GTGTTACATTACTGGGGCTT
930 SEQ ID NO:2565 -16.9 -34.9 86 -16.5 -1.4 -7.2
TTATTTTTTTCTTTGTTTTT
1605 SEQ ID NO: 2566 -16.9 -18.8 59.6 -1.9 0 0
TTTATTTTTTTCTTTGTTTT
1606 SEQ ID NO:2567 -16.9 -18.8 59.6 -1.9 0 0
AGAGCTATCATATCCTGCAT
1766 SEQ ID NO:2568 -16.9 -34.9 83.1 -17.1 -0.7 -5.2
CCTAACATGTTGAGCGTAGT
1813 SEQ ID NO: 2569 -16.9 -34.3 84.6 -16 0 -10.8
GATCAGGAGCAAAACACAGC
1992 SEQ ID NO:2570 -16.9 -36.1 85.4 -19.2 0 -5.2
TAATAATCAGATCAGGAGCA
2001 SEQ ID NO:2571 -16.9 -32.5 80.1 -15.6 0 -5.3
TGCAGGGTAGAGTCATTCTC
2028 SEQ ID NO: 2572 -16.9 -37.9 87.6 -19.9 -0.9 -9.2 ,
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecul Site oligo binding formation Duplex structure oligo oligo
AACCACTTCATGCATAGAAT
2300 SEQ ID NO: 2573 -16.9 -31.7 79.6 -13.8 0 -9.9
CGACTTTCTTTAAGGCAACC
2483 SEQ ID NO: 2574 -16.9 -32.6 82.6 -15.7 0 -5.4
AGCTATTAATTCGACTTTCT
2494 SEQ ID NO: 2575 -16.9 -28.8 75.9 -11.9 0 -4
CAAACTGATAGTTTATACAA
2518 SEQ ID NO:2576 -16.9 -26 72.4 -8.1 -0.6 -97
AGATGAAAACAATAAAAAAT
2561 SEQ ID NO:2577 -16.9 -21.9 64.4 -5 0 -0.4
ACATGTAGTGCGTATTTAAA
2591 SEQ ID NO:2578 -16.9 -28.6 76.8 -10.6 0 -10.2
AACTGCTTCTGTTGCCAAGT
2629 SEQ ID NO: 2579 -16.9 -35.4 86.2 -17.9 -0.3 -3.6
TGAAAAGTGATGACGACTCA
2648 SEQ ID NO:2580 -16.9 -33.3 81.1 -15.9 0 -8
TGAATAAAAAATTATGTAAG
3032 SEQ ID NO: 2581 -16.9 -21.2 64.1 -4.3 0 -4.2
CTGCCCATCTTAAACAGCTG
3062 SEQ ID NO: 2582 -16.9 -35.8 86.3 -18.2 0 -8.7
TTTTTTTTTTTGACAAGAAT
3209 SEQ ID NO:2583 -16.9 -21.1 63.5 -3.7 0 -7.8
TATCTGGAATCACAACTTTT
3495 SEQ ID NO: 2584 -16.9 -29.3 76.3 -12.4 0 -6.3
TCTCTCAGCTGTGTTACAGC
3519 SEQ ID NO:2585 -16.9 -37.6 87.8 -16.4 -4.1 -16.2
GATGGGAATGTGAAAATGGG
3619 SEQ ID NO:2586 -16.9 -33.8 82.1 -16.9 0 0
CCTTGAATAGAAATCAAATT
3900 SEQ ID NO:2587 -16.9 -25.7 70.7 -8.3 -0.2 -3.8
TTTACAATCATTTTAATAAA
4074 SEQ ID NO: 2588 -16.9 -20 61.6 -3.1 0 -0.3
TTTTACAATCATTTTAATAA
4075 SEQ ID NO:2589 -16.9 -20 61.6 -3.1 0 -0.3
TGATAAAACAATCTCATCTA
4127 SEQ ID NO: 2590 -16.9 -277 73.1 -10.8 0 -1.6
TCCATCTCCCTCTTCCCCTA
4267 SEQ ID NO:2591 -16.9 -41.5 92.3 -24.6 0 0
ACCACCTTCCTGTCTCCTGT
4458 SEQ ID NO: 2592 -16.9 -40.7 92 -23.8 0 0
CCTTTAAGGATGTAAGCACC
4475 SEQ ID NO:2593 -16.9 -33.8 84.3 -16 -0.6 -8.9
CACTATTTACATATTGCTGG
4593 SEQ ID NO:2594 -16.9 -29.5 77.7 -12.6 0 -17
CCTATGAGATTCTGCACTAT
4607 SEQ ID NO:2595 -16.9 -33.5 81.7 -16.6 0 -6
TAGAAAGGAATTAGTGTATT
4635 SEQ ID NO:2596 -16.9 -27.4 74.3 -10.5 0 -2.8
AGGAGCAGGAGGGAAATATA
18 SEQ ID NO: 2597 -16.8 -36.8 87.1 -20 0 -2.7
CCACCGTCCGCAGTTCCCGC
75 SEQ ID NO:2598 -16.8 -45 97.1 -28.2 0 -3.1 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecu Site oligo binding formation Duplex structure oligo oligo
GCCACCGTCCGCAGTTCCCG
76 SEQ ID NO:2599 -16.8 -45 97.1 -27.5 -0.4 -4.5
AGAAGCGACAGCCAGTGAGG
269 SEQ ID NO:2600 -16.8 -41 92 -23.1 -1 -7.1
GCCTTTTGGAAAATCAACCA
317 SEQ ID NO:2601 -16.8 -32 81.2 -14.1 -1 -4.6
TCTCCCATATACAGTCCCAT
388 SEQ ID NO: 2602 -16.8 -37.1 86.7 -20.3 0 -07
CATCACTTTTGTTTCTGTCT
407 SEQ ID NO: 2603 -16.8 -30 77.4 -13.2 0 0
GCAAACAGTTTTCATCTATC
786 SEQ ID NO: 2604 -16.8 -29.4 76.5 -12.6 0 -4
TCTTCTTTTTCTGTTTTCAC
958 SEQ ID NO:2605 -16.8 -27.6 74.1 -10.8 0 0
CTCCAGAGGTACTCACACCA
1098 SEQ ID NO:2606 -16.8 -39.9 90.8 -20.8 -2.3 -8.3
CTATTCCAATTTTCGGAACC
1210 SEQ ID NO: 2607 -16.8 -30.7 78.8 -11.4 -2.5 -7.8
CACCAGGCAGAGTTTGGGAG
1382 SEQ ID NO: 2608 -16.8 -40.3 91.7 -18.4 -5.1 -15.3
GCTATCATATCCTGCATATA
1763 SEQ ID NO: 2609 -16.8 -32 79.7 -14.5 -0.5 -4.5
CCTCCAAGTTGAGTCTGGAA
1787 SEQ ID NO:2610 -16.8 -37.2 87.3 -17.3 -2.6 -14.1
ATTAATAATCAGATCAGGAG
2003 SEQ ID NO:2611 -16.8 -29 74.9 -12.2 0 -5.3
CTTTTCCTAGCTCTTTGATG
2211 SEQ ID NO:2612 -16.8 -31.4 79.9 -14.6 0 -3.8
CAGATGAAAACAATAAAAAA
2562 SEQ ID NO:2613 -16.8 -22.9 66.7 -6.1 0 -2
ATCCTAACTATACAGGGGGG
2870 SEQ ID NO: 2614 -16.8 -37.9 89.5 -17.9 -3.2 -10.8
CTATCCTAACTATACAGGGG
2872 SEQ ID NO:2615 -16.8 -34.7 85.4 -157 -2.2 -6.5
ATGCTATCCTAACTATACAG
2875 SEQ ID NO:2616 -16.8 -31.4 80.2 -14.6 0 -17
CAGGTTTCCATGCATAAATC
2899 SEQ ID NO:2617 -16.8 -31.6 79.6 -14.3 0 -7.9
AAGCGTAGTTCACTAAATAT
3001 SEQ ID NO:2618 -16.8 -28.8 76.8 -12 0 -5.4
GTACAATAACTTGAATAAAA
3043 SEQ ID NO:2619 -16.8 -23.6 687 -6.8 0 -5
TGTGGTCTTCTGATAGCTAA
3167 SEQ ID NO: 2620 -16.8 -34.4 83.9 -15.4 -2.2 -10.4
TTGTGGTCTTCTGATAGCTA
3168 SEQ ID NO: 2621 -16.8 -34.4 83.9 -15.4 -2.2 -8.5
TATTTTTTAAATTAGTCTTT
3359 SEQ ID NO:2622 -16.8 -19.8 61.7 -3 0 -1.8
TTATTTTTTAAATTAGTCTT
3360 SEQ ID NO: 2623 -16.8 -19.8 61.7 -3 0 -1.9
GTCTGACATACAGTTCTAAA
3700 SEQ ID NO: 2624 -16.8 -30.9 79 -13.4 -0.4 -8.1 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
CTTATTTTACACTATACAAG 4098 SEQ ID NO: 2625 -16.8 -24.9 71.5 -8.1 0 -0.4
TCATGATAAAACAATCTCAT 4130 SEQ ID NO: 2626 -16.8 -27.5 72.2 -10.7 0 -6.2
GCAACCTATGAGATTCTGCA 4611 SEQ ID NO:2627 -16.8 -35.4 84.4 -17.6 -0.9 -5.9
AGTGTATTATTGGCAACCTA 4623 SEQ ID NO: 2628 -16.8 -31.8 81.6 -15 0 -4.9
TCACATCTCCCCTCTCCTGA 192 SEQ ID NO: 2629 -16.7 -41.5 91.1 -24.8 0 -1.1
ACCTATTGAGGTTTGCAATG 507 SEQ ID NO:2630 -16.7 -32.4 81.6 -13.4 -2.2 -11.9
TGTGGGGGCAGCAGACACAG 569 SEQ ID NO: 2631 -16.7 -43.8 95.9 -23.4 -2.5 -15.4
AATGTTGCTGTTCTGAAGAT 621 SEQ ID NO:2632 -167 -31.4 78.9 -147 0 -5.3
GGGCAGTGTTACATTACTGG 935 SEQ ID NO: 2633 -16.7 -36.1 87.3 -14.8 -4.6 -10.9
TAGCGGCATGCTGGGCAGTT 1546 SEQ ID NO: 2634 -16.7 -42 94.6 -20.4 -2.3 -17.9
CAGGATTTTCAGAGGTTTCT 1659 SEQ ID NO: 2635 -16.7 -32.5 80.8 -15.3 0 -7.8
AAGTTGAGTCTGGAACAGAG 1782 SEQ ID NO: 2636 -16.7 -34.9 84.1 -16.6 -1.4 -10.6
GATCCTCCAAGTTGAGTCTG 1790 SEQ ID NO: 2637 -167 -36.5 85.2 -18.9 -0.8 -8.2
GAAACATCCAGGAGTACTGC 1920 SEQ ID NO: 2638 -16.7 -36.5 86.2 -18.1 -1.7 -8.7
ATACTCTTCATAAGATACCT 2102 SEQ ID NO: 2639 -167 -30.3 77.7 -13.6 0 -4.3
TCCTTAGGAACTGAAGAGAG 2146 SEQ ID NO:2640 -16.7 -35.4 84.9 -17.5 -0.3 -10.3
GAATCCAAGAGTTTTGTCAG 2284 SEQ ID NO: 2641 -16.7 -31.4 79.1 -14.2 0 -7.7
GTATCTGATTGGTGATGATT 2397 SEQ ID NO:2642 -16.7 -31.2 77.6 -13 -1.4 -5.4
CTTATTAAGGCAGTCACTTT 2460 SEQ ID NO: 2643 -16.7 -30.2 79.2 -13.5 0 -6.2
CTTTAAGGCAACCATTCTTA 2476 SEQ ID NO:2644 -167 -30.1 79.3 -13.4 0 -5.2
TTCTTTAAGGCAACCATTCT 2478 SEQ ID NO: 2645 -16.7 -31.2 80.2 -14.5 0 -5.4
TGTTGCCAAGTCTTGGCCCT 2620 SEQ ID NO: 2646 -16.7 -40.1 92.4 -16.6 -4.3 -21.8
AAGTGATGACGACTCAACTG 2644 SEQ ID NO: 2647 -167 -34.3 82.7 -17.1 -0.1 -5.5
ATTTCACCATCTACTCTCCC 2673 SEQ ID NO: 2648 -16.7 -35.6 84.5 -18.9 0 0
AGAGAAAGTTCATCACACAG 2791 SEQ ID NO: 2649 -16.7 -32.7 80.6 -16 0 -7
AAGTATGAAGAGAAAGTTCA 2799 SEQ ID NO: 2650 -167 -29.4 76.4 -12.7 0 -5.6 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecu Site oligo binding formation Duplex structure oligo oligo
GGGGATACACCAACAGAAAG
2852 SEQ ID NO:2651 -16.7 -36.1 86.5 -17.4 -2 -7.1
GGGGGGGGATACACCAACAG
2856 SEQ ID NO:2652 -16.7 -43 95.3 -22.9 -3.4 -7.3
GCCATCAGGTTAGAAGCACC
3145 SEQ ID NO:2653 -16.7 -38.6 89.3 -20.5 -1.3 -5.3
AAAATATGAGCTTTTTTTTT
3227 SEQ ID NO:2654 -16.7 -21.4 64.3 -4.2 0 -7.5
GGTCCTGAAAGACAAATAGT
3449 SEQ ID NO:2655 -16.7 -32.8 81.8 -14.2 -1.9 -5.6
TACAAACTACTTCAAAAGGT
3466 SEQ ID NO: 2656 -16.7 -28.9 77.7 -12.2 0 -1.9
TTATACAAACTACTTCAAAA
3469 SEQ ID NO:2657 -16.7 -24.6 70.5 -7.9 0 -0.5
TTTAAGAAGTTATACAAACT
3478 SEQ ID NO:2658 -167 -24.6 70.6 -7.9 0 -2
GTGAGAGGAATTACTTTGTC
3549 SEQ ID NO:2659 -16.7 -31.9 80 -13.6 -1.6 -6.4
ATGGGAATGTGAAAATGGGT
3618 SEQ ID NO:2660 -16.7 -33.6 82.3 -16.9 0 0
TGTACCATCAGGAAAGATTA
3653 SEQ ID NO: 2661 -16.7 -32.1 80.5 -14.5 -0.8 -4.2
AATCACAAAAATCAGTAGCT
3682 SEQ ID NO:2662 -16.7 -29.1 76.3 -12.4 0 -3
ATAGGTTATCCATCAGCATT
4181 SEQ ID NO:2663 -16.7 -32.2 80.2 -14.3 -1.1 -5.2
GGTGCCTTTAAGGATGTAAG
4479 SEQ ID NO:2664 -167 -33.8 84.3 -16 -0.7 -9.8
CTTCTGAAGCTTCTGTTGTC
4548 SEQ ID NO:2665 -16.7 -33.7 82.5 -14.8 0 -12.5
GAGTTAATGATTCTTTGGAG
138 SEQ ID NO:2666 -16.6 -29.5 76.4 -11.7 -07 -10
CTCCTGAGCAAGCACACTGC
179 SEQ ID NO: 2667 -16.6 -40.5 91.3 -23.3 -0.3 -5.7
ATTGCTTACTGAGCCTTTTG
329 SEQ ID NO: 2668 -16.6 -32.2 81.8 -13.8 -1.4 -11.4
GTGGGAATCCCAGGTCATTT
429 SEQ ID NO:2669 -16.6 -37.3 87.1 -15.9 -3.5 -17.7
GTTTCCCCCGAGGAAAGGCT
463 SEQ ID NO:2670 -16.6 -40.8 92.8 -20.8 -3.4 -11.4
ACTCACACCATGAACAGAAA
1088 SEQ ID NO:2671 -16.6 -33.7 82.4 -17.1 0 -4.1
GTACTCACACCATGAACAGA
1090 SEQ ID NO:2672 -16.6 -35.4 84.6 -18.8 0 -4.1
AAAACTTTACAGCTTCCACA
1444 SEQ ID NO:2673 -16.6 -30.9 80.4 -14.3 0 -4
ACATAGGTAATTGTGCTGTC
1484 SEQ ID NO:2674 -16.6 -33 82.3 -15 -1.3 -6.6
GCACATAGGTAATTGTGCTG
1486 SEQ ID NO:2675 -16.6 -33.9 83.8 -10.8 -6.5 -14.9
TGTTTTGTTACCAGGATTTT
1670 SEQ ID NO: 2676 -16.6 -28.6 76.7 -10.8 -1.1 -7.1 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
TATTGTTTTGTTACCAGGAT 1673 SEQ ID NO:2677 -16.6 -29.2 77.3 -11.5 -1 -6.2
AGCTATCATATCCTGCATAT 1764 SEQ ID NO: 2678 -16.6 -32.8 80.6 -15.3 -0.7 -4.5
ATGATTTCAGCTAACATCTC 2383 SEQ ID NO: 2679 -16.6 -31 77.8 -14.4 0 -6.1
TTTAGAGTTTATTTGGGAAA 3094 SEQ ID NO: 2680 -16.6 -26.7 737 -10.1 0 -3.8
CAAATAATTTGGCCACCTTG 3915 SEQ ID NO:268l -16.6 -30.8 80 -12.7 0 -11
TACACAAATAATTTGGCCAC 3919 SEQ ID NO: 2682 -16.6 -30.2 79.1 -12.1 0 -11
ATATTAGAAGTTCCATATTT 3965 SEQ ID NO: 2683 -16.6 -25.3 70.2 -87 0 -4.9
AAATATATTAGAAGTTCCAT 3969 SEQ ID NO: 2684 -16.6 -25.3 70.2 -8.7 0 -5.1
CCTTCCCTTCCCAGATTAGT 4032 SEQ ID NO: 2685 -16.6 -37.4 88.1 -20.8 0 -1.1
TCCCTCTTCCCCTAGAGCAA 4261 SEQ ID NO:2686 -16.6 -41.1 92.8 -21.8 -2.7 -7.8
CTGTTTACATACTTTACATA 4442 SEQ ID NO: 2687 -16.6 -26.2 73.3 -9.6 0 -4.4
AAGGATGTAAGCACCACCTT 4470 SEQ ID NO: 2688 -16.6 -35.9 86.8 -17 -2.3 -9.1
GGCTGTATGTGAAAGTAACC 4510 SEQ ID NO: 2689 -16.6 -34 84.2 -16.9 -0.2 -4.9
GAGGGCTGTATGTGAAAGTA 4513 SEQ ID NO: 2690 -16.6 -35.4 85.6 -18.3 -0.2 -4.9
CCTCATTTTATTTGGAAATA 4671 SEQ ID NO:269l -16.6 -25.8 71.5 -9.2 0 -3.9
TTAATGATTCTTTGGAGTCC 135 SEQ ID NO: 2692 -16.5 -307 78.1 -10.4 -3.8 -14.2
GGAAAATCAACCAAAAGTCT 310 SEQ ID NO: 2693 -16.5 -29.9 77.7 -12.7 -0.5 -2.9
CCATCACTTTTGTTTCTGTC 408 SEQ ID NO:2694 -16.5 -31.2 79.2 -14.7 0 0
AAATGTTGCTGTTCTGAAGA 622 SEQ ID NO: 2695 -16.5 -31.2 79.1 -14.7 0 -3.6
CAGAGGAGAAAGCAAACAGT 797 SEQ ID NO: 2696 -16.5 -34.6 84.3 -18.1 0 -2.7
CCCGCCAGAGGAGAAAGCAA 802 SEQ ID NO: 2697 -16.5 -40.8 92.4 -22.7 -1.5 -5.8
TGACATTAAAAATAGGCTTC 1173 SEQ ID NO: 2698 -16.5 -28.2 75.5 -11.7 0 -5
TAGGTAATTGTGCTGTCCTT 1481 SEQ ID NO: 2699 -16.5 -33.9 84.3 -16.5 -0.8 -8.1
GAGTCTGGAACAGAGCTATC 1777 SEQ ID NO: 2700 -16.5 -37 85.9 -18.3 -2 -12
ATTTGGTCATCCAGGTGTAA 1891 SEQ ID NO: 2701 -16.5 -33.9 83.2 -13.4 -4 -11.2
TCATTTGGTCATCCAGGTGT 1893 SEQ ID NO: 2702 -16.5 -36.2 85.4 -13.1 -6.6 -16.4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target imolecular molecul Site oligo binding formation Duplex structure oligo oligo
GTCCTTAGGAACTGAAGAGA
2147 SEQ ID NO:2703 -16.5 -35.5 84.9 -17.5 -0.5 -11
CGTCCTTAGGAACTGAAGAG
2148 SEQ ID NO: 2704 -16.5 -35.5 85.3 -17.5 -0.5 -11
CTCTTTGATGTAGGTCATTC
2201 SEQ ID NO: 2705 -16.5 -32 79.6 -14.1 -07 -10.7
TAATTCGACTTTCTTTAAGG
2488 SEQ ID NO:2706 -16.5 -27.3 74.3 -10.8 0 -4.6
TGTTTAGAGTTTATTTGGGA
3096 SEQ ID NO: 2707 -16.5 -29.2 77.4 -12.7 0 -3.8
CCTGTACCATCAGGAAAGAT
3655 SEQ ID NO:2708 -16.5 -35.3 84.5 -14.1 -47 -11.8
GGATGTCTGACATACAGTTC
3704 SEQ ID NO:2709 -16.5 -34.6 82.7 -15.8 -0.4 -12.8
CTATGGCACACATTAGGGAT
3742 SEQ ID NO:2710 -16.5 -35.4 85.2 -17.9 -0.9 -4
ATTGCCAAACTTCTGAAGCT
4557 SEQ ID NO:2711 -16.5 -33.3 82.7 -16.3 0 -8.3
CAACCTATGAGATTCTGCAC
4610 SEQ ID NO: 2712 -16.5 -34.2 82.8 -17.7 0 -6
CCGCCACCGTCCGCAGTTCC
78 SEQ ID NO:2713 -16.4 -45 97.1 -27.7 -0.7 -4.6
ACATCTCCCCTCTCCTGAGC
190 SEQ ID NO: 2714 -16.4 -42.5 92.5 -24.5 -1.6 -5.4
TCCCATATACAGTCCCATTG
386 SEQ ID NO:2715 -16.4 -35.6 85.1 -19.2 0 -1.7
CTCCCATATACAGTCCCATT
387 SEQ ID NO:2716 -16.4 -35.6 85.2 -19.2 0 -0.5
AGATCTCCATTATCCTTAAT
901 SEQ ID NO:2717 -16.4 -30.2 76.7 -13.8 0 -5.2
CTCTTGCTTAATTACCCCAG
998 SEQ ID NO: 2718 -16.4 -33.7 84.2 -17.3 0 -2.3
GGCTTGAATAGCCATTAGAA
1299 SEQ ID NO: 2719 -16.4 -33 82.3 -10.9 -57 -14.4
CTTCCAGCACATAGGTAATT
1492 SEQ ID NO: 2720 -16.4 -32.8 82.3 -16.4 0 -2.7
AGGTTCATTCCAGCCTGAAG
1576 SEQ ID NO:2721 -16.4 -37.1 87.2 -17.3 -3.4 -9.2
GCAGGAACTATTGTTTTGTT
1681 SEQ ID NO:2722 -16.4 -29.9 78.5 -11.7 -1.8 -7
GAGCTATCATATCCTGCATA
1765 SEQ ID NO: 2723 -16.4 -34.1 82.2 -16.8 -0.7 -5.2
CATGCAGGGTAGAGTCATTC
2030 SEQ ID NO: 2724 -16.4 -36.6 85.8 -19.7 -0.2 -5.9
CTAGCTCTTTGATGTAGGTC
2205 SEQ ID NO: 2725 -16.4 -34.4 83.8 -18 0 -5.6
AGAGTTTTGTCAGTTGATAA
2277 SEQ ID NO: 2726 -16.4 -29.2 76.4 -11 -1.6 -11
ATGCATAGAATCCAAGAGTT
2291 SEQ ID NO:2727 -16.4 -31.9 79.5 -15.5 0 -6.1
ATATTTTGGTATCTGATTGG
2405 SEQ ID NO: 2728 -16.4 -28.5 75 -12.1 0 -5.4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target ιmolecular molecul Site oligo binding formation Duplex structure oligo oligo
CCTCTACAGGACAAACTGAT
2529 SEQ ID NO: 2729 -16.4 -35.2 84.7 -16 -2.8 -6.8
AAAATTTCATCCAGCCAACT
2827 SEQ ID NO: 2730 -16.4 -31 79.2 -14.6 0 -1.3
TTTGTGGTCTTCTGATAGCT
3169 SEQ ID NO:2731 -16.4 -34 83.2 -15.4 -2.2 -6.6
AATATGAGCTTTTTTTTTTT
3225 SEQ ID NO:2732 -16.4 -21.4 64.3 -5 0 -6.2
AAATATGAGCTTTTTTTTTT
3226 SEQ ID NO:2733 -16.4 -21.4 64.3 -5 0 -6.4
ATTTTTTAAATTAGTCTTTT
3358 SEQ ID NO:2734 -16.4 -19.4 60.7 -3 0 -1.9
TATAGCCATTGCAAAAATAG
3411 SEQ ID NO: 2735 -16.4 -28.2 76 -11.1 -0.5 -6.7
GTTATACAAACTACTTCAAA
3470 SEQ ID NO: 2736 -16.4 -25.9 72.8 -9.5 0 -1.7
GAGAGGAATTACTTTGTCTG
3547 SEQ ID NO:2737 -16.4 -31.8 79.9 -13 -2.4 -9.8
TTTTTGCCATATTAGAGATT
3589 SEQ ID NO:2738 -16.4 -27.1 73.3 -10.7 0 -1.9
AGCTGAGCTTTCCTGTACCA
3666 SEQ ID NO: 2739 -16.4 -38.4 89.9 -20.6 0 -10.7
AATACCAATATGATATATAT
4011 SEQ ID NO: 2740 -16.4 -24 67.6 -7.6 0 -4.8
CAGCAACCTTCACTGCACAC
4297 SEQ ID NO: 2741 -16.4 -37.9 88.7 -19.2 -2.3 -5.9
CTTCCTCTGTAATCTCACTG
4328 SEQ ID NO: 2742 -16.4 -34.2 82.9 -17.8 0 -0.6
TGCCAAACTTCTGAAGCTTC
4555 SEQ ID NO:2743 -16.4 -34.6 84.4 -17.1 0 -10.1
GATAGAAAGGAATTAGTGTA
4637 SEQ ID NO: 2744 -16.4 -28.9 76.2 -12.5 0 -2.8
CTCCCCTCTCCTGAGCAAGC
186 SEQ ID NO: 2745 -16.3 -43.2 94.1 -25.5 -1.3 -5.1
CACAGCAGTGGATGCTGAAC
554 SEQ ID NO: 2746 -16.3 -38.3 88.2 -15.2 -6.8 -16.3
GGGCAGCAGACACAGCAGTG
564 SEQ ID NO: 2747 -16.3 -42.7 94.1 -25.3 -1 -6.7
AAAGCAAACAGTTTTCATCT
789 SEQ ID NO: 2748 -16.3 -28.5 757 -11.5 -0.4 -3
TCCTTAATTTTGGGTTTAGT
889 SEQ ID NO: 2749 -16.3 -28.9 77.6 -11.2 -1.3 -8
ATTTTATTACCAATTATATT
1060 SEQ ID NO: 2750 -16.3 -20.5 62.5 -4.2 0 -0.7
TCACACCATGAACAGAAATG
1086 SEQ ID NO: 2751 -16.3 -32.6 80.4 -15.8 -0.1 -4.3
GGAATTGGTGGAATGACATT
1186 SEQ ID NO: 2752 -16.3 -32.7 80.5 -15.5 -0.7 -6.2
TGGCCTGCTGAATTCCTTTT
1623 SEQ ID NO: 2753 -16.3 -35.6 86 -18.8 0 -8.3
TATCCTGCATATAACACTTC
1756 SEQ ID NO: 2754 -16.3 -31.3 79.4 -15 0 -4.4 . .
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecul Site oligo binding formation Duplex structure oligo oligo
CTAACATGTTGAGCGTAGTC
1812 SEQ ID NO: 2755 -16.3 -33.4 82.8 -15.7 0 -10.8
TAGGGTCATTTGGTCATCCA
1898 SEQ ID NO:2756 -16.3 -36.5 86.3 -17.6 -2.6 -10.8
AACTCAGAGGAAACATACAG
2071 SEQ ID NO:2757 -16.3 -32.8 81.5 -16.5 0 -7.2
GGTCATTCTAATTTCATCAA
2189 SEQ ID NO: 2758 -16.3 -28.5 74.4 -12.2 0 -3.4
TGGTCTTATCCAAAAATGTT
2346 SEQ ID NO:2759 -16.3 -29 76.4 -10.9 -1.8 -6
TAATAGCTATAAAAGGCACA
2956 SEQ ID NO:2760 -16.3 -29.7 78.9 -11.1 -2.3 -9.8
TTTTGTGGTCTTCTGATAGC
3170 SEQ ID NO:2761 -16.3 -32.8 81.6 -15.4 -1 -4.5
CCAGCTTTCTTGCCATATAG
3426 SEQ ID NO: 2762 -16.3 -33.9 83.8 -16.5 -1 -4
AACTTCCTCTGTAATCTCAC
4330 SEQ ID NO:2763 -16.3 -33.1 81.6 -16.8 0 -0.6
AGGCAGAGGATAACTTCCTC
4341 SEQ ID NO: 2764 -16.3 -37.8 87.8 -15.8 -5.7 -14
GTAAGCACCACCTTCCTGTC
4464 SEQ ID NO:2765 -16.3 -38.5 89.6 -22.2 0 -2.7
GTATGTGAAAGTAACCCGCT
4506 SEQ ID NO:2766 -16.3 -34.3 84.8 -17.5 -0.2 -3.4
TCCAAAATACTGAAAATAAC
4732 SEQ ID NO:2767 -16.3 -25.8 71.8 -9.5 0 -0.4
TCTCTCCCATATACAGTCCC
390 SEQ ID NO:2768 -16.2 -38.4 88.2 -22.2 0 -0.7
CAAATGTTGCTGTTCTGAAG
623 SEQ ID NO:2769 -16.2 -30.9 79.1 -147 0 -3.4
GACAAAACCAGATCTCCATT
910 SEQ ID NO: 2770 -16.2 -32.8 80.7 -16.6 0 -6.5
TAATTACCCCAGGGGTGCAG
990 SEQ ID NO:2771 -16.2 -39.5 92.2 -16.4 -4.3 -21.9
GTTTCTCTTGCTTAATTACC
1002 SEQ ID NO: 2772 -16.2 -29.3 77.6 -13.1 0 -2.3
TAATTGTGCTGTCCTTCCAC
1477 SEQ ID NO: 2773 -16.2 -35 84.9 -18.8 0 -3.2
GTAATTGTGCTGTCCTTCCA
1478 SEQ ID NO: 2774 -16.2 -35 84.9 -18.8 0 -3.8
TCCTTTTATTTTTTTCTTTG
1610 SEQ ID NO:2775 -16.2 -22.6 66.8 -6.4 0 0
TGCAATCACTTGCCGCCCTC
1832 SEQ ID NO:2776 -16.2 -40.9 92 -22.8 -1.9 -6.6
ACTCAGAGGAAACATACAGC
2070 SEQ ID NO:2777 -16.2 -35.3 847 -19.1 0 -7.2
CAATAGTTAAGGAGATTTTC
2320 SEQ ID NO:2778 -16.2 -27.2 737 -11 0 -6.8
GTCTTATCCAAAAATGTTTG
2344 SEQ ID NO: 2779 -16.2 -26.6 72.9 -10.4 0 -3.6
ATGGTCTTATCCAAAAATGT
2347 SEQ ID NO:2780 -16.2 -29.2 76.2 -10.9 -2.1 -6.8 . .
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecul Site oligo binding formation Duplex structure oligo oligo
ACTGTGAAAAAAAGTATGAA
2810 SEQ ID NO:2781 -16.2 -26.5 72.8 -10.3 0 -2.6
AACTGTGAAAAAAAGTATGA
2811 SEQ ID NO: 2782 -16.2 -26.5 72.8 -10.3 0 -3.7
GCCCATCTTAAACAGCTGTA
3060 SEQ ID NO: 2783 -16.2 -35.1 85.8 -16.2 0 -13.6
GTTTAGAGTTTATTTGGGAA
3095 SEQ ID NO: 2784 -16.2 -28 75.8 -11.8 0 .-3.9
CATATAGCCATTGCAAAAAT
3413 SEQ ID NO:2785 -16.2 -28 75 -11.1 -0.5 -6.7
TCCTGTACCATCAGGAAAGA
3656 SEQ ID NO:2786 -16.2 -36.6 86.4 -14.1 -6.3 -15
AATTTCTTGTGGCTTAGTAA
3884 SEQ ID NO:2787 -16.2 -29 77.5 -12.8 0 -5
TTACAATCATTTTAATAAAT
4073 SEQ ID NO:2788 -16.2 -20.2 61.6 -4 0 -1.5
TACAAGCTATTTTACAATCA
4084 SEQ ID NO:2789 -16.2 -27.2 74.2 -11 0 -5
TAACATGTCATGATAAAACA
4137 SEQ ID NO: 2790 -16.2 -27.3 73.2 -9.2 0 -11.9
TGACCCCTACAAAAAATATA
4157 SEQ ID NO: 2791 -16.2 -29.5 78.1 -13.3 0 -0.9
TTCAACTGCTGGGGGAGGGC
4527 SEQ ID NO:2792 -16.2 -43.9 96.2 -26.3 -1.1 -10.2
CCTTTTGGAAAATCAACCAA
316 SEQ ID NO: 2793 -16.1 -29.5 77.7 -11.8 -1.5 -4.9
GCTGCTGCGCATTGCTTACT
339 SEQ ID NO: 2794 -16.1 -38.5 89.9 -19.5 -1.5 -13.9
CACTTTTGTTTCTGTCTCTC
404 SEQ ID NO:2795 -16.1 -31.3 79.3 -15.2 0 0
AGGTCATTTCCCATCACTTT
418 SEQ ID NO:2796 -16.1 -33.7 82.4 -16.9 -0.4 -3.4
AGAAGAAAACTCCAAATCCT
716 SEQ ID NO: 2797 -16.1 -31.3 79.3 -15.2 0 -0.3
CTCCCGCCAGAGGAGAAAGC
804 SEQ ID NO: 2798 -16.1 -42.3 93.3 -22.2 -4 -10.8
CTTTTTCTGTTTTCACTTGG
954 SEQ ID NO:2799 -16.1 -28.2 76 -12.1 0 -0.5
TTTTATTACCAATTATATTT
1059 SEQ ID NO:2800 -16.1 -20.3 62.5 -4.2 0 -0.7
ATCATTCCTTCCAGCACATA
1499 SEQ ID NO:2801 -16.1 -34.3 82.7 -18.2 0 -2.7
CTGCAATCACTTGCCGCCCT
1833 SEQ ID NO -.2802 -16.1 -40.6 92.1 -22.6 -1.9 -6.6
ACTGCTGCAATCACTTGCCG
1837 SEQ ID NO: 2803 -16.1 -38.3 89.1 -20.3 -1.9 -7.9
TTGATGTAGGTCATTCTAAT
2197 SEQ ID NO:2804 -16.1 -29.9 76.8 -12.2 -0.9 -10.9
GAAGCAATAGTTAAGGAGAT
2324 SEQ ID NO:2805 -16.1 -30.9 78.9 -14.8 0 -2.7
TATTTTGGTATCTGATTGGT
2404 SEQ ID NO:2806 -16.1 -29.6 77 -13.5 0 -5.4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
AAAACCTCTACAGGACAAAC
2533 SEQ ID NO: 2807 -16.1 -32.4 82.1 -14.7 -1.5 -7.3
GATGAAAACAATAAAAAATA
2560 SEQ ID NO: 2808 -16.1 -21.1 63.5 -5 0 -0.4
AATTTCACCATCTACTCTCC
2674 SEQ ID NO:2809 -16.1 -33.2 81.4 -17.1 0 0
TTCTAATTTCTGGGATATAT
2703 SEQ ID NO:2810 -16.1 -27.9 74 -10.8 -0.9 -6.8
GGTCTTCTGATAGCTAAGTG
3164 SEQ ID NO:2811 -16.1 -34.4 83.8 -17.4 -0.8 -7.5
ACAAAATATGAGCTTTTTTT
3229 SEQ ID NO:2812 -16.1 -23.9 687 -7.8 0 -6.4
ACTTCAAAAGGTCCTGAAAG
3458 SEQ ID NO:2813 -16.1 -32.1 81.2 -15.1 -0.7 -9
TAGCCCTTCCCTTCCCAGAT
4036 SEQ ID NO: 2814 -16.1 -41 92.7 -24.9 0 -1.3
ACTTCCTCTGTAATCTCACT
4329 SEQ ID NO:2815 -16.1 -34.3 83.2 -18.2 0 -0.6
CTTTAGTGCAAGGGGAGATT
4422 SEQ ID NO:2816 -16.1 -34.8 85 -17.4 0 -10.5
CATACTTTACATACTTTAGT
4435 SEQ ID NO:2817 -16.1 -26.2 73.4 -10.1 0 -2.8
TGAAAGTAACCCGCTATTAG
4501 SEQ ID NO:2818 -16.1 -32.1 82.3 -15.5 -0.2 -3.1
AGGGCTGTATGTGAAAGTAA
4512 SEQ ID NO:2819 -16.1 -33.9 84.3 -17.3 -0.2 -4.9
GCTGAACTCTTGGGGTTCTC
541 SEQ ID NO:2820 -16 -38.5 88.7 -17 -4.9 -19
TAATTTTGGGTTTAGTGTCC
885 SEQ ID NO: 2821 -16 -30.2 79.4 -12.6 -1.5 -3
TTAATTTTGGGTTTAGTGTC
886 SEQ ID NO: 2822 -16 -27.8 75.8 -11.8 0 -2.3
CCCCAGAGAAGTCAAGTTGT
1250 SEQ ID NO: 2823 -16 -37 87.5 -21 0 -3.8
CGATGATGCAATCATTCCTT
1509 SEQ ID NO: 2824 -16 -32.2 78.9 -14 -1.7 -12.4
AACTGAAGAGAGAAGCAGTA
2138 SEQ ID NO:2825 -16 -34.1 83.4 -16.4 -1.7 -9.1
CGTATTTAAAACAAAACAAC
2581 SEQ ID NO: 2826 -16 -23.1 68.7 -7.1 0 -2.9
GACTCAACTGCTTCTGTTGC
2634 SEQ ID NO: 2827 -16 -36 85.6 -18.2 -1.8 -5.8
CTAATTTCTGGGATATATTA
2701 SEQ ID NO: 2828 -16 -26.8 73 -10.8 0 -6.6
CACATATTAAGGTTTCTAAT
2716 SEQ ID NO:2829 -16 -26.1 72.4 -10.1 0 -4
TTACGTCCACATATTAAGGT
2723 SEQ ID NO:2830 -16 -31.2 80.4 -15.2 0 -3
AGTATGAAGAGAAAGTTCAT
2798 SEQ ID NO:2831 -16 -29.6 76.2 -12.9 -0.5 -5.6
AATCAAAAATGCTATCCTAA
2883 SEQ ID NO: 2832 -16 -27.1 73.3 -11.1 0 -17 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CTGGACTAGGTGCTCTATAC
3305 SEQ ID NO: 2833 -16 -37 87.6 -20.3 -0.3 -8.1
CAAAAATAGGGCGTTAGGTA
3400 SEQ ID NO: 2834 -16 -31.8 82.5 -15.8 0 -6.9
TGGATGTCTGACATACAGTT
3705 SEQ ID NO: 2835 -16 -34.3 82.9 -15.5 -0.8 -13.7
CTCTGTAATCTCACTGGGTC
4324 SEQ ID NO: 2836 -16 -36.7 86 -19.7 -0.9 -7
GGCAGAGGATAACTTCCTCT
4340 SEQ ID NO:2837 -16 -37.8 87.8 -14.2 -7.6 -18
GCTTCTGTTGTCATTCAACT
4540 SEQ ID NO: 2838 -16 -32.5 80.8 -14.2 -2.3 -7.6
ACGCAGAAGGAGCAGGAGGG
25 SEQ ID NO: 2839 -15.9 -43.3 94.9 -26.3 -1 -3.3
GTGAAACTGCTTTGGACAGA
363 SEQ ID NO:2840 -15.9 -34.7 84.3 -17.6 -1 -9.3
TGGGAATCCCAGGTCATTTC
428 SEQ ID NO: 2841 -15.9 -37.5 87 -17.1 -3.3 -17
TTTCTTCCAAAAGCTTTAAG
486 SEQ ID NO:2842 -15.9 -27.3 75 -10.8 0 -8.6
GTTTGCAATGCTTTCTTCCA
497 SEQ ID NO: 2843 -15.9 -32.1 81 -15.7 0 -7.6
ACTCCAAATCCTGCAAAATG
708 SEQ ID NO: 2844 -15.9 -32.2 80.7 -16.3 0 -4.9
AGTTTCCTTCCAAAAGGAAT
834 SEQ ID NO: 2845 -15.9 -31 79.5 -10.6 -4.5 -12.5
GCTTGCAGTCCTCATTCGAG
852 SEQ ID NO:2846 -15.9 -37.8 87.2 -21 -07 -7.9
GCTTCTGATCCTGCTGTTGA
1158 SEQ ID NO: 2847 -15.9 -37.5 86.8 -21.1 -0.1 -6.4
AGGTGGTCCTGTTGTTGCTG
1364 SEQ ID NO: 2848 -15.9 -39 90.5 -21.5 -1.5 -6.2
CACTTGCCGCCCTCCTAACA
1826 SEQ ID NO:2849 -15.9 -40.8 93.3 -24.9 0 -3.9
ATCACTTGCCGCCCTCCTAA
1828 SEQ ID NO:2850 -15.9 -40 91.7 -24.1 0 -3.9
AATAATCAGATCAGGAGCAA
2000 SEQ ID NO:2851 -15.9 -32.1 79.4 -16.2 0 -5.3
TTTGGAAGCAATAGTTAAGG
2328 SEQ ID NO:2852 -15.9 -30.1 79.3 -13.4 -0.6 -27
CCAAAAATGTTTGGAAGCAA
2337 SEQ ID NO:2853 -15.9 -29.3 77.6 -10.4 -3 -11
TATCTGATTGGTGATGATTT
2396 SEQ ID NO:2854 -15.9 -29.9 75.8 -12.7 -1.2 -5.4
GTTTTTTGATATTTCCATTT
2427 SEQ ID NO: 2855 -15.9 -23.1 66.8 -7.2 0 -2.3
GAAGTTTTTTGATATTTCCA
2430 SEQ ID NO: 2856 -15.9 -25.6 70.9 -8.3 -1.3 -6.6
AAAACAACAAAACCTCTACA
2541 SEQ ID NO: 2857 -15.9 -28.5 76.9 -12.6 0 0
GTTGCCAAGTCTTGGCCCTC
2619 SEQ ID NO: 2858 -15.9 -40.4 91.9 -17.7 -4.3 -21.8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TAGCTGCCCATCTTAAACAG
3065 SEQ ID NO:2859 -15.9 -35 85.7 -19.1 0 -5.5
ACAGATGATTGATTAATGTT
3112 SEQ ID NO: 2860 -15.9 -27.3 72.3 -10.8 0 -8.4
CAAAATATGAGCTTTTTTTT
3228 SEQ ID NO: 2861 -15.9 -22.6 66.7 -6.7 0 -6.8
GTAGTTATTTTTTAAATTAG
3364 SEQ ID NO: 2862 -15.9 -20 63.4 -4.1 0 -1.9
TCAAAAGGTCCTGAAAGACA
3455 SEQ ID NO: 2863 -15.9 -33.6 82.8 -15.8 -1.9 -8.7
GTTTAGTGAGAGGAATTACT
3554 SEQ ID NO: 2864 -15.9 -30.8 79.2 -12.9 -2 -6.6
ATATCTGAAACTATAACTAA
3995 SEQ ID NO: 2865 -15.9 -25.6 71.1 -9.7 0 -1.7
ATTTTACAATCATTTTAATA
4076 SEQ ID NO: 2866 -15.9 -20.2 61.6 -4.3 0 -0.3
TATTTTACAATCATTTTAAT
4077 SEQ ID NO: 2867 -15.9 -20.2 61.6 -4.3 0 -0.3
ATAGAAAGGAATTAGTGTAT
4636 SEQ ID NO: 2868 -15.9 -27.6 74.2 -11.7 0 -2.8
GGATAGAAAGGAATTAGTGT
4638 SEQ ID NO: 2869 -15.9 -30.9 79 -15 0 -2.8
TGGCAGAGGAGCCGCTCGCC
97 SEQ ID NO:2870 -15.8 -47.3 99.1 -257 -5.8 -18.2
TTGAGAGTGAAACTGCTTTG
369 SEQ ID NO: 2871 -15.8 -32.2 81 -14 -2.4 -7.6
TGTCTCTCCCATATACAGTC
392 SEQ ID NO: 2872 -15.8 -36.1 85 -20.3 0 -1
CTTGGCACCTATTCCAATTT
1218 SEQ ID NO: 2873 -15.8 -32.3 817 -15 ' -1.4 -6.5
GTTGCTGTTGAGGAGCTGGA
1351 SEQ ID NO: 2874 -15.8 -39.5 90.2 -21.2 -2.5 -11.7
GATTTTCAGAGGTTTCTTGT
1656 SEQ ID NO: 2875 -15.8 -30.2 77.6 -13.9 0 -7.6
AACAGAGCTATCATATCCTG
1769 SEQ ID NO: 2876 -15.8 -33.5 81.7 -17.7 0 -4.8
TCATTAATAATCAGATCAGG
2005 SEQ ID NO: 2877 -15.8 -29 74.8 -13.2 0 -5.2
AGGAAACATACAGCATGTGT
2064 SEQ ID NO: 2878 -15.8 -33.8 83.1 -17.1 -0.7 -8.9
GTCATTCTAATTTCATCAAA
2188 SEQ ID NO: 2879 -15.8 -26.1 707 -10.3 0 0
CAATACTCATGGTCTTATCC
2355 SEQ ID NO: 2880 -15.8 -32.3 80.2 -16.5 0 -5.2
GGCAACCATTCTTATTAAGG
2470 SEQ ID NO: 2881 -15.8 -31.5 80.8 -157 0 -5.9
ACTTTCTTTAAGGCAACCAT
2481 SEQ ID NO: 2882 -15.8 -31 80.3 -15.2 0 -5.3
ACCATCTACTCTCCCATCAC
2668 SEQ ID NO: 2883 -15.8 -38.1 87.4 -22.3 0 0
TAACTTGAATAAAAAATTAT
3037 SEQ ID NO: 2884 -15.8 -20 61.6 -4.2 0 -0.3 kcal/mol kcal/mol degC kcal/mol kcal/mol kcal/mol Intra Inter
Target total duplex Tmof target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
TTACAGCTGGTTATCTGGAA
3506 SEQ ID NO:2885 -15.8 -34.1 84.2 -16.6 -1.4 -11.1 GTGTTACAGCTGGTTATCTG
3509 SEQ ID NO:2886 -15.8 -34 84 -16.6 0 -11.1 ATGGATGTCTGACATACAGT
3706 SEQ ID NO:2887 -15.8 -34.5 82.6 -15.5 -1.1 -14.5 TTTCAACAGTGAAGAAATTC
3781 SEQ ID NO:2888 -15.8 -27.8 73.9 -10.9 -1 -8.1 ATATATCTGAAACTATAACT
3997 SEQ ID NO:2889 -15.8 -25.8 71 -10 0 -1.4 GGATAACTTCCTCTGTAATC
4334 SEQ ID NO: 2890 -15.8 -32.4 80.6 -14.2 -2.4 -7.2 ACTAAACCTGCGCTGACAGA
4396 SEQ ID NO: 2891 -15.8 -37.6 88.8 -20.4 -1.3 -7.5 TAGATGGTGCCTTTAAGGAT
4484 SEQ ID NO: 2892 -15.8 -34.2 84 -16.8 -07 -11 AAGCTTCTGTTGTCATTCAA
4542 SEQ ID NO: 2893 -15.8 -31.2 79.1 -15.4 0 -4.5 TTGGCAACCTATGAGATTCT
4614 SEQ ID NO: 2894 -15.8 -34.1 83.1 -18.3 0 -4 TAAATAATTTCTCCAAAATA
4743 SEQ ID NO: 2895 -15.8 -22.6 66.1 -6.8 0 -0.3 TTTCCTTCCAAAAGGAATGA
832 SEQ ID NO:2896 -15.7 -31.2 79.3 -11 -4.5 -12.5 TAGAAAAAACTGTTCGACCA
1284 SEQ ID NO:2897 -15.7 -30.5 79.1 -14.8 0 -4.4 TTTTATTTTTTTCTTTGTTT
1607 SEQ ID NO:2898 -15.7 -18.8 59.6 -3.1 0 0 TGTGGTAACGTTGCAGGAAC
1693 SEQ ID NO:2899 -15.7 -36.2 87.4 -17.3 -1.1 -14.6 TCATATCCTGCATATAACAC
1759 SEQ ID NO: 2900 -157 -31.5 79.1 -15.8 0 -4.5 ACATGTTGAGCGTAGTCATG
1809 SEQ ID NO:2901 -15.7 -34.4 83.3 -17.6 -0.8 -9.5 TCCTAACATGTTGAGCGTAG
1814 SEQ ID NO: 2902 -15.7 -34.5 84.4 -17.4 0 -10.8 CCATTTCACTGCTGCAATCA
1844 SEQ ID NO:2903 -157 -34.7 83.7 -18.5 0 -7.9 TTGTCTATATGATCTCCACC
1958 SEQ ID NO:2904 -15.7 -33.8 81.8 -18.1 0 -4.2 TAACTCAGAGGAAACATACA
2072 SEQ ID NO:2905 -15.7 -32 80.7 -16.3 0 -7.2 CTCTTGGCTCTTCAGACCGT
2165 SEQ ID NO:2906 -15.7 -38.8 89.1 -21.2 -1.9 -9.4 AGGCAACCATTCTTATTAAG
2471 SEQ ID NO: 2907 -15.7 -30.3 79 -14.6 0 -3.9 GTATTTAAAACAAAACAACA
2580 SEQ ID NO:2908 -15.7 -22.8 67.9 -7.1 0 -2.8 TTCTGTTGCCAAGTCTTGGC
2623 SEQ ID NO: 2909 -15.7 -36.8 87.6 -15.1 -3.5 -20.2 ACATATTAAGGTTTCTAATT
2715 SEQ ID NO:2910 -15.7 -24.9 70.4 -9.2 0 -4 ,
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecular Site oligo binding formation Duplex structure oligo oligo
CACTTGTAAACTTTTTTTCA
2917 SEQ ID NO:2911 -15.7 -24.8 71.1 -9.1 0 -2
AATAAAAAATTATGTAAGAA
3030 SEQ ID NO:2912 -157 -20 61.6 -4.3 0 -4.2
ATGTTTAGAGTTTATTTGGG
3097 SEQ ID NO: 2913 -157 -27.9 75.5 -12.2 0 -3.6
CACAGATGATTGATTAATGT
3113 SEQ ID NO:2914 -15.7 -28.5 74.1 -12.1 0 -8.8
TTTTTTTTTTTTTTGACAAG
3212 SEQ ID NO:2915 -157 -19.4 61.4 -3.7 0 -1.9
AGAAGTTCCATATTTTTAAT
3960 SEQ ID NO:2916 -157 -24.7 69.5 -8.5 -0.2 -4.9
TCTGAAACTATAACTAAATA
3992 SEQ ID NO: 2917 -15.7 -25.4 71.2 -97 0 -1
TATCTGAAACTATAACTAAA
3994 SEQ ID NO:2918 -157 -25.4 71.2 -97 0 -1.7
CCCTTCCCTTCCCAGATTAG
4033 SEQ ID NO:2919 -15.7 -38.5 89.5 -22.8 0 -1.1
AACATGTCATGATAAAACAA
4136 SEQ ID NO: 2920 -15.7 -26.9 72.4 -9.2 0 -12.2
TATTGCCAAACTTCTGAAGC
4558 SEQ ID NO: 2921 -15.7 -32.5 81.8 -16.3 0 -8.3
TATTGGCAACCTATGAGATT
4616 SEQ ID NO: 2922 -15.7 -32 80.4 -16.3 0 -4.9
CTTACTGAGCCTTTTGGAAA
325 SEQ ID NO: 2923 -15.6 -32.2 82.1 -15.5 -1 0
CAGGTCATTTCCCATCACTT
419 SEQ ID NO: 2924 -15.6 -34.9 83.8 -18.6 -0.4 -3.4
TTGCAATGCTTTCTTCCAAA
495 SEQ ID NO:2925 -15.6 -30.8 79.3 -14.7 0 -7.6
TACTCACACCATGAACAGAA
1089 SEQ ID NO:2926 -15.6 -34.1 83.1 -18.5 0 -4.1
AAAGGGATGCTGTATTCATG
1137 SEQ ID NO: 2927 -15.6 -32.8 81.2 -14.6 -0.4 -13.4
CAAGTTAAGACTCCATAATG
1426 SEQ ID NO: 2928 -15.6 -29.4 77.1 -13.8 0 -5
TTTTTCGAGCTTCCAGGTTC
1590 SEQ ID NO:2929 -15.6 -33.9 83.4 -15.4 -2.9 -10.1
TAACGTTGCAGGAACTATTG
1688 SEQ ID NO:2930 -15.6 -31.8 81.4 -16.2 0 -5.8
ACCAGGGTAGGGGTGAGTTG
1711 SEQ ID NO: 2931 -15.6 -41.7 94.3 -24.5 -1.5 -8
ATCCAGGAGTACTGCAGTAG
1915 SEQ ID NO: 2932 -15.6 -37.8 88.5 -18.3 -1.7 -16
TAGTTAAGGAGATTTTCAAC
2317 SEQ ID NO:2933 -15.6 -28.3 757 -11.8 0 -9.8
TTCTTATTAAGGCAGTCACT
2462 SEQ ID NO: 2934 -15.6 -31.7 80.9 -16.1 0 -7.2
TTCTGGGATATATTAACTAA
2696 SEQ ID NO:2935 -15.6 -27.9 75 -12.3 0 -6.4
CATATTAAGGTTTCTAATTT
2714 SEQ ID NO:2936 -15.6 -23.6 68.4 -8 0 -4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
TAATGTTTAGAGTTTATTTG 3099 SEQ ID NO: 2937 -15.6 -23.5 68.5 -7.9 0 -3.8
ACCCATTTTCACACAGATGA 3124 SEQ ID NO: 2938 -15.6 -33.8 82.3 -16.9 -1.2 -4.9
AGGTCCTGAAAGACAAATAG 3450 SEQ ID NO: 2939 -15.6 -32.7 81.7 -15.2 -1.9 -7.8
TCTGGAATCACAACTTTTAA 3493 SEQ ID NO: 2940 -15.6 -29.1 76.5 -13.5 0 -5.5
CATCAGGAAAGATTAACCAA 3648 SEQ ID NO: 2941 -15.6 -30.4 78.1 -14.8 0 -3.4
GTAGTTTTACAAACATGGAT 3720 SEQ ID NO: 2942 -15.6 -28.1 75.5 -12.5 0 -6.9
AACTCTATGGCACACATTAG 3746 SEQ ID NO: 2943 -15.6 -32.9 82.2 -17.3 0 -4.2
GTGTTAAACTCTATGGCACA 3752 SEQ ID NO: 2944 -15.6 -32.8 82.5 -12.8 -4.4 -11.6
TAAGTTTTGAGTTTACAGAA 3858 SEQ ID NO: 2945 -15.6 -26.7 73.8 -10.5 -0.3 -2.6
TCTTATTTTACACTATACAA 4099 SEQ ID NO: 2946 -15.6 -25.2 71.2 -9.6 0 -0.4
CCCCTCTCCTGAGCAAGCAC 184 SEQ ID NO: 2947 -15.5 -43 94.2 -25.9 -1.6 -5.9
GCTTTCTTCCAAAAGCTTTA 488 SEQ ID NO:2948 -15.5 -29.8 78.7 -11 -3.3 -8.8
TCAAATGTTGCTGTTCTGAA 624 SEQ ID NO: 2949 -15.5 -31.2 79.1 -157 0 -2.8
GAGGTGGTCCTGTTGTTGCT 1365 SEQ ID NO:2950 -15.5 -39.3 90.5 -21.9 -1.9 -7.6
AGGATTTTCAGAGGTTTCTT 1658 SEQ ID NO:295l -15.5 -31.3 79.3 -15.3 0 -7.6
CATTTGGTCATCCAGGTGTA 1892 SEQ ID NO: 2952 -15.5 -35.1 84.6 -13.1 -6.5 -16.2
AGTAGGGTCATTTGGTCATC 1900 SEQ ID NO: 2953 -15.5 -35.4 84.6 -17.9 -2 -10.8
AGTACTGCAGTAGGGTCATT 1908 SEQ ID NO: 2954 -15.5 -36.4 87.4 -17 -0.1 -16
CATCCAGGAGTACTGCAGTA 1916 SEQ ID NO:2955 -15.5 -37.8 88.4 -19.3 -1.7 -14.1
ATAATCAGATCAGGAGCAAA 1999 SEQ ID NO: 2956 -15.5 -32.1 79.4 -16.6 0 -5.3
GCAGGGTAGAGTCATTCTCT 2027 SEQ ID NO: 2957 -15.5 -37.9 87.6 -20.2 -2.2 -9.4
ACATACAGCATGTGTTTACA 2059 SEQ ID NO: 2958 -15.5 -31.6 80.6 -14.9 -0.8 -9.8
ATTCTTATTAAGGCAGTCAC 2463 SEQ ID NO:2959 -15.5 -30.7 79 -15.2 0 -7.2
CAAAACCTCTACAGGACAAA 2534 SEQ ID NO: 2960 -15.5 -32.3 82 -15.7 -1 -6.8
TATTTAAAACAAAACAACAG 2579 SEQ ID NO: 2961 -15.5 -22.7 67.7 -7.2 0 -2.2
GTAGTGCGTATTTAAAACAA 2587 SEQ ID NO: 2962 -15.5 -27.2 75.3 -10.8 0 -9.8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target ιmolecular molecu Site oligo binding formation Duplex structure oligo oligo
CGACTCAACTGCTTCTGTTG
2635 SEQ ID NO: 2963 -15.5 -35 84.5 -18.1 -1.3 -4.9
TTTCACCATCTACTCTCCCA
2672 SEQ ID NO: 2964 -15.5 -36.6 86.4 -21.1 0 0
GTTTCTAATTTCTGGGATAT
2705 SEQ ID NO: 2965 -15.5 -28.6 75.3 -11.5 -1.5 -6.8
GAGAAAGTTCATCACACAGA
2790 SEQ ID NO: 2966 -15.5 -33 80.6 -17.5 0 -7
CAAGCGTAGTTCACTAAATA
3002 SEQ ID NO:2967 -15.5 -29.8 78.7 -13.8 -0.2 -5.6
TACAATAACTTGAATAAAAA
3042 SEQ ID NO:2968 -15.5 -22.3 66.2 -6.8 0 -1.3
AAGCAGATATATACAAAATA
3241 SEQ ID NO:2969 -15.5 -25.5 71 -10 0 -3.5
GCACAACCTATATAATTCTC
3266 SEQ ID NO:2970 -15.5 -30.1 77.9 -14.6 0 -3.4
ATATAGCCATTGCAAAAATA
3412 SEQ ID NO:2971 -15.5 -27.2 74 -11.1 -0.3 -6.7
AAAAGGTCCTGAAAGACAAA
3453 SEQ ID NO:2972 -15.5 -30.9 79.6 -13.5 -1.9 -9
TTTTAAGAAGTTATACAAAC
3479 SEQ ID NO:2973 -15.5 -23.4 68.8 -7.9 0 -3.4
ATCTGAAACTATAACTAAAT
3993 SEQ ID NO: 2974 -15.5 -25.2 70.3 -97 0 -1.7
TACAATCATTTTAATAAATT
4072 SEQ ID NO:2975 -15.5 -20.2 61.6 -4.7 0 -1.5
TCATTCTTATTTTACACTAT
4103 SEQ ID NO:2976 -15.5 -25.2 70.3 -9.7 0 0
ATCATTCTTATTTTACACTA
4104 SEQ ID NO:2977 -15.5 -25.2 70.3 -9.7 0 0
CTGCACACAGGACCAGTCTC
4285 SEQ ID NO:2978 -15.5 -40.8 91.2 -24 -1 -10
TTGCCAAACTTCTGAAGCTT
4556 SEQ ID NO:2979 -15.5 -33.1 83 -17.1 0 -8.3
TTATTGGCAACCTATGAGAT
4617 SEQ ID NO:2980 -15.5 -32 80.4 -16 -0.2 -5.7
ATTAGTGTATTATTGGCAAC
4626 SEQ ID NO:2981 -15.5 -28.4 76.2 -12.9 0 -4.9
AGAAAGGAATTAGTGTATTA
4634 SEQ ID NO:2982 -15.5 -27.4 74.3 -11.9 0 -2.8
CAAAAAGCAAATGATTGTTT
4769 SEQ ID NO:2983 -15.5 -24.7 69.9 -7.6 -1.5 -5.8
AAGTCCATCACATCTCCCCT
199 SEQ ID NO: 2984 -15.4 -38.8 88.4 -23.4 0 -0.6
GGTTTGCAATGCTTTCTTCC
498 SEQ ID NO:2985 -15.4 -33.3 827 -17.4 0 -7.6
CCGGTAAAATGAGAGGCTTG
867 SEQ ID NO:2986 -15.4 -35.1 85.4 -197 0 -6.5
CACCATGAACAGAAATGGCA
1083 SEQ ID NO:2987 -15.4 -34.7 83.8 -16.1 -3.2 -8.7
GGTACTCACACCATGAACAG
1091 SEQ ID NO:2988 -15.4 -36.3 86.2 -18.8 -2.1 -6.2 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
AGCGGCATGCTGGGCAGTTT
1545 SEQ ID NO:2989 -15.4 -41.6 93.9 -21.3 -1.8 -17.9
AGGGTAGGGGTGAGTTGTGG
1708 SEQ ID NO:2990 -15.4 -417 94.3 -26.3 0 -2.7
TTCATCAAATAGCTCTTGGC
2177 SEQ ID NO:2991 -15.4 -32.9 81.4 -17.5 0 -6
GGAAGCAATAGTTAAGGAGA
2325 SEQ ID NO:2992 -15.4 -33.1 82.5 -17.7 0 -27
TTTCTAATTTCTGGGATATA
2704 SEQ ID NO:2993 -15.4 -27.7 74.1 -11.1 -1.1 -6.8
CAACTGTGAAAAAAAGTATG
2812 SEQ ID NO:2994 -15.4 -26.2 72.9 -10.8 0 -3.7
AATAACTTGAATAAAAAATT
3039 SEQ ID NO: 2995 -15.4 -19.6 60.7 -4.2 0 -0.2
AGATTTGAGTCAATTTTTGT
3184 SEQ ID NO: 2996 -15.4 -26.5 71.8 -9.5 -0.7 -11
ACAACCTATATAATTCTCCA
3264 SEQ ID NO: 2997 -15.4 -30 77.8 -14.6 0 -3.4
CACAACCTATATAATTCTCC
3265 SEQ ID NO:2998 -15.4 -30 77.8 -14.6 0 -3.4
CTAGCCATTTTTGCCATATT
3596 SEQ ID NO:2999 -15.4 -30.2 78.9 -14.8 0 -2.2
ACAGATGGGAATGTGAAAAT
3622 SEQ ID NO:3000 -15.4 -31.5 78.8 -15 -1 -4.3
AGTTTTACAAACATGGATGT
3718 SEQ ID NO:3001 -15.4 -28.9 76.4 -12.8 0 -9
GAAGTTCCATATTTTTAATA
3959 SEQ ID NO: 3002 -15.4 -23.9 68.5 -8.5 0 -4.5
TATTAGAAGTTCCATATTTT
3964 SEQ ID NO: 3003 -15.4 -25.1 70.3 -97 0 -47
AAAATATATTAGAAGTTCCA
3970 SEQ ID NO: 3004 -15.4 -25.1 70.3 -9.7 0 -5.1
ATTTTAATAAATTGCATGTA
4065 SEQ ID NO:3005 -15.4 -22.4 66 -6.5 0 -7.6
AACTTCTGAAGCTTCTGTTG
4550 SEQ ID NO:3006 -15.4 -32.2 81 -14.6 0 -12.5
TTTACATATTGCTGGTACCT
4588 SEQ ID NO:3007 -15.4 -31.8 81.6 -15.1 0 -10.6
AACATGTCCTCATTTTATTT
4678 SEQ ID NO: 3008 -15.4 -26.8 72.5 -11.4 0 -6.2
ATAACATTCAAAAAGCATTC
4710 SEQ ID NO: 3009 -15.4 -25.6 70.8 -10.2 0 -2.7
CATCAGTGAATATCAACTCT
116 SEQ ID NO: 3010 -15.3 -31 77.6 -15.7 0 -67
CTGAGCCTTTTGGAAAATCA
321 SEQ ID NO:3011 -15.3 -32.2 80.9 -16 -0.8 -6.2
TTTGCAATGCTTTCTTCCAA
496 SEQ ID NO:3012 -15.3 -30.8 79.3 -15 0 -7.6
AGAGGAGAAAGCAAACAGTT
796 SEQ ID NO: 3013 -15.3 -33.4 82.9 -18.1 0 -27
GCCAGAGGAGAAAGCAAACA
799 SEQ ID NO: 3014 -15.3 -37 87.5 -21.7 0 -3.1 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GAGTTTCCTTCCAAAAGGAA
835 SEQ ID NO:3015 -15.3 -32.3 81.3 -13.1 -3.9 -10.8
TGGAATGACATTAAAAATAG
1178 SEQ ID NO:3016 -15.3 -25.9 71.4 -10.6 0 -5
AGAAAAAACTGTTCGACCAG
1283 SEQ ID NO:3017 -15.3 -31.3 80 -14.8 -0.3 -10.3
CTTGAATAGCCATTAGAAAA
1297 SEQ ID NO:3018 -15.3 -28.1 75.4 -12.8 0 -2.8
TTGCTGTTGAGGAGCTGGAT
1350 SEQ ID NO:3019 -15.3 -38.4 88.6 -21.2 -1.5 -11.7
GAGCTTCCAGGTTCATTCCA
1584 SEQ ID NO:3020 -15.3 -37.4 87.1 -17.8 -4.3 -12.9
TCTTTGTTTTTCGAGCTTCC
1596 SEQ ID NO:3021 -15.3 -31.5 80 -16.2 0 -6.6
CTTTTATTTTTTTCTTTGTT
1608 SEQ ID NO:3022 -15.3 -20 62.4 -47 0 0
TTGTGGTAACGTTGCAGGAA
1694 SEQ ID NO:3023 -15.3 -34.9 85.9 -16.4 -1.7 -14.6
CATTAATAATCAGATCAGGA
2004 SEQ ID NO:3024 -15.3 -29 74.8 -13.7 0 -5.2
GGAAACATACAGCATGTGTT
2063 SEQ ID NO:3025 -15.3 -32.6 81.5 -15.4 -1.9 -9.8
CTCTTCATAAGATACCTGAA
2099 SEQ ID NO:3026 -15.3 -31.1 78.7 -15.8 0 -4.3
TGCATAGAATCCAAGAGTTT
2290 SEQ ID NO:3027 -15.3 -31.7 797 -16.4 0 -37
GGTATCTGATTGGTGATGAT
2398 SEQ ID NO:3028 -15.3 -33.6 80.8 -16.8 -1.4 -5.4
AACTGATAGTTTATACAATA
2516 SEQ ID NO:3029 -15.3 -25.4 71.1 -10.1 0 -5.8
GCGTATTTAAAACAAAACAA
2582 SEQ ID NO:3030 -15.3 -24.3 70.6 -9 0 -37
TAAAAGGCACAACTTCCCTT
2947 SEQ ID NO:3031 -15.3 -33.2 84.1 -17.2 -0.4 -5
TTGTTAAAACCAGACAGTAA
2973 SEQ ID NO:3032 -15.3 -28.9 77.6 -12.6 -0.9 -4.2
CAGATGATTGATTAATGTTT
3111 SEQ ID NO:3033 -15.3 -26 70.5 -10.7 0 -7.4
CTCTATACCAGTTAGGACTG
3293 SEQ ID NO:3034 -15.3 -34.5 84.6 -17.4 -1.6 -11.2
TACTTCAAAAGGTCCTGAAA
3459 SEQ ID NO:3035 -15.3 -31.3 80.3 -15.1 -0.7 -9
CTACTTCAAAAGGTCCTGAA
3460 SEQ ID NO:3036 -15.3 -32.5 81.9 -16.5 0 -9
TACAGCTGGTTATCTGGAAT
3505 SEQ ID NO: 3037 -15.3 -34.3 83.9 -17.4 -1.2 -11.1
CTCTATGGCACACATTAGGG
3744 SEQ ID NO:3038 -15.3 -36.4 86.8 -17.9 -3.2 -8.4
TTAGAAGTTCCATATTTTTA
3962 SEQ ID NO:3039 -15.3 -24.9 70.4 -8.5 -1 -5
TGAATACCAATATGATATAT
4013 SEQ ID NO:3040 -15.3 -26.1 70.7 -10.8 0 -4.8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GTCATGATAAAACAATCTCA
4131 SEQ ID NO: 3041 -15.3 -28.6 74.3 -12.5 0 -9.2
AATCATATAGGTTATCCATC
4187 SEQ ID NO: 3042 -15.3 -29.4 75.7 -12.9 -1.1 -5.2
CTGCGCTGACAGACTCACTG
4389 SEQ ID NO: 3043 -15.3 -39.9 90.1 -23.5 -1 -7.9
TACTTTAGTGCAAGGGGAGA
4424 SEQ ID NO: 3044 -15.3 -36.3 877 -19.5 -0.1 -11
TGAAACTGCTTTGGACAGAT
362 SEQ ID NO: 3045 -15.2 -33.6 82.5 -17.6 -0.5 -8.8
CTTTAAGTCTGTTTCCCCCG
473 SEQ ID NO: 3046 -15.2 -34.9 85.8 -19.7 0 -2.1
CTCCTTCTCTGTGGGGGCAG
578 SEQ ID NO: 3047 -15.2 -42.9 94.3 -23.1 -4.6 -12.5
TCCAGAGGTACTCACACCAT
1097 SEQ ID NO: 3048 -15.2 -38.9 89.3 -21.4 -2.3 -8.5
GTCATAGTGGTACATCTGTC
1118 SEQ ID NO: 3049 -15.2 -34.8 .83.5 -187 -0.3 -9.3
TGTTGCTGTTGAGGAGCTGG
1352 SEQ ID NO: 3050 -15.2 -39.2 90.3 -21.5 -2.5 -11.7
TTACAGCTTCCACAAGTTAA
1438 SEQ ID NO: 3051 -15.2 -31.3 81.2 -14.9 -1.1 -57
AAACTTTACAGCTTCCACAA
1443 SEQ ID NO: 3052 -15.2 -30.9 80.4 -15.7 0 -4
GTGCTGTCCTTCCACTGCTC
1472 SEQ ID NO: 3053 -15.2 -40.8 91.3 -24.4 -1.1 -4.6
CCCATTTCACTGCTGCAATC
1845 SEQ ID NO: 3054 -15.2 -35.9 85.3 -20.2 0 -7.9
TCCAGGAGTACTGCAGTAGG
1914 SEQ ID NO:3055 -15.2 -40 91.7 -20.9 -1.7 -16
AGAGCAAATGCCATAAGAAA
1936 SEQ ID NO:3056 -15.2 -31.4 79.8 -15.4 -0.6 -6.1
ACCTCTACAGGACAAACTGA
2530 SEQ ID NO:3057 -15.2 -36.3 86.7 -18.3 -2.8 -7.3
TAATTTCTGGGATATATTAA
2700 SEQ ID NO: 3058 -15.2 -25.6 71 -10.4 0 -6.8
AAATCAAAAATGCTATCCTA
2884 SEQ ID NO:3059 -15.2 -27.1 73.3 -11.9 0 -1.7
ACGAACTAGCTGCCCATCTT
3071 SEQ ID NO:3060 -15.2 -37.7 88.6 -22.5 0 -6.3
AACTACTTCAAAAGGTCCTG
3462 SEQ ID NO: 3061 -15.2 -32.3 82.1 -16.4 0 -9
ATAACATGTCATGATAAAAC
4138 SEQ ID NO:3062 -15.2 -26.3 71.4 -9.1 0 -12.2
GACCCCTACAAAAAATATAT
4156 SEQ ID NO:3063 -15.2 -28.5 76.3 -13.3 0 -1.4
CCCCTAGAGCAAACTGTTTG
4253 SEQ ID NO: 3064 -15.2 -35.6 86.8 -19.2 -1.1 -8.1
AACTAAACCTGCGCTGACAG
4397 SEQ ID NO: 3065 -15.2 -36.1 87.2 -20.4 -0.2 -7.5
TTTAGTGCAAGGGGAGATTG
4421 SEQ ID NO: 3066 -15.2 -34.8 85 -19.6 0 -5.1 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CCTGTTTACATACTTTACAT
4443 SEQ ID NO:3067 -15.2 -28.2 76.3 -13 0 -4.4
GTATTATTGGCAACCTATGA
4620 SEQ ID NO:3068 -15.2 -31 79.6 -15.8 0 -4.9
CCAAAATACTGAAAATAACA
4731 SEQ ID NO:3069 -15.2 -25.5 71.6 -10.3 0 -0.4
AATTTTGGGTTTAGTGTCCG
884 SEQ ID NO:3070 -15.1 -31.3 80.8 -13 -3.2 -6.4
GTAAACTGTGCCCAGTTTCT
1016 SEQ ID NO:3071 -15.1 -34.6 85.4 -15.8 -3.4 -15
TTGGCACCTATTCCAATTTT
1217 SEQ ID NO: 3072 -15.1 -31.1 80.1 -14.6 -1.3 -5.7
CAAGTTGAGTCTGGAACAGA
1783 SEQ ID NO: 3073 -15.1 -34.9 84.1 -18.9 -0.6 -8.7
CTCAGAGGAAACATACAGCA
2069 SEQ ID NO: 3074 -15.1 -35.2 84.6 -20.1 0 -6.4
AATTCGACTTTCTTTAAGGC
2487 SEQ ID NO: 3075 -15.1 -29.4 77.3 -14.3 0 -5.4
ACAGAAAGTCTAGAAAATTT
2840 SEQ ID NO:3076 -15.1 -26.7 72.8 -11.1 0 -7.5
AGGCACAACTTCCCTTTTCT
2943 SEQ ID NO:3077 -15.1 -35.5 86.4 -20.4 0 -2.8
CCATCTTAAACAGCTGTACA
3058 SEQ ID NO: 3078 -15.1 -327 82.4 -14.9 0 -13.6
CTAAGTGCCATCAGGTTAGA
3151 SEQ ID NO:3079 -15.1 -35.3 85.6 -18.2 -2 -6
AAAGATTAACCAATTGGTGA
3641 SEQ ID NO: 3080 -15.1 -29 76.4 -10.2 -2.2 -15.6
GTTAAACTCTATGGCACACA
3750 SEQ ID NO:3081 -15.1 -32.8 82.5 -17.2 -0.1 -4.2
GAAAGTAACCCGCTATTAGA
4500 SEQ ID NO: 3082 -15.1 -32.4 82.3 -17.3 0 -4.1
GGAGTTAATGATTCTTTGGA
139 SEQ ID NO: 3083 -15 -30.7 78.1 -14.1 -1.2 -11
CATCTCCCCTCTCCTGAGCA
189 SEQ ID NO: 3084 -15 -42.4 92.4 -25.8 -1.6 -5.4
TTTGGACAGATCTGGCTGCT
353 SEQ ID NO: 3085 -15 -38.4 88.6 -20.3 -1.5 -14.4
TACTGGGGCTTGACAAAACC
921 SEQ ID NO:3086 -15 -36.9 88.6 -21.4 0 -77
ATTACTGGGGCTTGACAAAA
923 SEQ ID NO:3087 -15 -33.4 837 -18.4 0 -5.9
TTGTTGCTGTTGAGGAGCTG
1353 SEQ ID NO: 3088 -15 -36.8 87.3 -19.3 -2.5 -11.7
CCTTCCAGCACATAGGTAAT
1493 SEQ ID NO: 3089 -15 -35.2 85.5 -19.6 -0.3 -6.7
TTTATCGATGATGCAATCAT
1514 SEQ ID NO: 3090 -15 -29.2 74.3 -12.5 -1.5 -11
TTCCTTTTATTTTTTTCTTT
1611 SEQ ID NO: 3091 -15 -21.4 64.4 -6.4 0 0
TTTTGTTACCAGGATTTTCA
1668 SEQ ID NO: 3092 -15 -28.8 76.6 -13.8 0 -4.3 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecu Site oligo binding formation Duplex structure oligo oligo
TCACTTGCCGCCCTCCTAAC
1827 SEQ ID NO:3093 -15 -41.1 93.2 -26.1 0 -3.9
GGGTAGAGTCATTCTCTGCT
2024 SEQ ID NO: 3094 -15 -37.9 87.6 -14.9 -8 -19
TACGTCCACATATTAAGGTT
2722 SEQ ID NO: 3095 -15 -31.2 80.4 -16.2 0 -3
AAATTTCATCCAGCCAACTG
2826 SEQ ID NO: 3096 -15 -32.2 807 -17.2 0 -2.6
ATAGCCATTGCAAAAATAGG
3410 SEQ ID NO: 3097 -15 -30.2 78.9 -14.5 -0.5 -67
AGCCATTTTTGCCATATTAG
3594 SEQ ID NO: 3098 -15 -30.2 78.9 -15.2 0 -1.8
AAGTTTTGAGTTTACAGAAA
3857 SEQ ID NO:3099 -15 -26.3 73 -10.1 -1.1 -4.3
AATCATTCTTATTTTACACT
4105 SEQ ID NO:3100 -15 -24.8 69.4 -9.8 0 0
ACATGTCATGATAAAACAAT
4135 SEQ ID NO: 3101 -15 -27.1 72.3 -10.1 0 -12.2
CTTTGACCCCTACAAAAAAT
4160 SEQ ID NO: 3102 -15 -29.7 78.6 -14.7 0 -2.4
CACCACCTTCCTGTCTCCTG
4459 SEQ ID NO: 3103 -15 -40.6 91.4 -25.6 0 0
AAGGAATTAGTGTATTATTG
4631 SEQ ID NO: 3104 -15 -26.1 72.4 -11.1 0 -4.3
ACAAAAACATGTCCTCATTT
4683 SEQ ID NO: 3105 -15 -287 75.6 -13.7 0 -6.2
AACAAAAACATGTCCTCATT
4684 SEQ ID NO:3106 -15 -28.7 75.6 -13.7 0 -6.2
GATAAACAACTTAGCTTGTG
46 SEQ ID NO: 3107 -14.9 -29.1 77.4 -13.7 -0.2 -6.8
CTGTATTCATGTCATAGTGG
1128 SEQ ID NO: 3108 -14.9 -32.1 80.1 -16.6 -0.3 -6.8
GGAACAGAGCTATCATATCC
1771 SEQ ID NO:3109 -14.9 -35 83.3 -19.1 -0.9 -7.3
ACTTGCCGCCCTCCTAACAT
1825 SEQ ID NO:3110 -14.9 -39.8 91.9 -24.9 0 -3.9
TCTAATTTCATCAAATAGCT
2183 SEQ ID NO: 3111 -14.9 -27.4 73.2 -12.5 0 -2.9
CAAAAATGTTTGGAAGCAAT
2336 SEQ ID NO: 3112 -14.9 -27.1 73.8 -11.4 -0.6 -8
AAAGTCTAGAAAATTTCATC
2836 SEQ ID NO: 3113 -14.9 -25.9 70.9 -10.5 0 -7.5
GATACACCAACAGAAAGTCT
2849 SEQ ID NO: 3114 -14.9 -32.9 81.6 -18 0 -3.3
TACACTTGTAAACTTTTTTT
2919 SEQ ID NO: 3115 -14.9 -23.8 69.8 -8.9 0 -5.5
TAGCTATAAAAGGCACAACT
2953 SEQ ID NO: 3116 -14.9 -31.6 81.9 -14.2 -2.5 -6.3
AACCCATTTTCACACAGATG
3125 SEQ ID NO:3117 -14.9 -32.3 80.7 -16.9 -0.1 -2.6
CCATCAGGTTAGAAGCACCA
3144 SEQ ID NO: 3118 -14.9 -37.3 87.9 -20.8 -1.6 -6 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GCAGATATATACAAAATATG
3239 SEQ ID NO: 3119 -14.9 -25.7 71.1 -10.8 0 -3.5
GCCATTGCAAAAATAGGGCG
3407 SEQ ID NO: 3120 -14.9 -34.8 85.4 -16.9 -3 -11.4
TTCAAAAGGTCCTGAAAGAC
3456 SEQ ID NO: 3121 -14.9 -32.4 81.2 -16.6 -0.6 -9
ACAAATAATTTGGCCACCTT
3916 SEQ ID NO: 3122 -14.9 -30.9 80.2 -14.5 0 -11
AGAGGATAACTTCCTCTGTA
4337 SEQ ID NO: 3123 -14.9 -34.6 84 -11.7 -8 -18.4
TTCTGTTGTCATTCAACTGC
4538 SEQ ID NO:3124 -14.9 -32.5 80.8 -15.3 -2.3 -7.6
TGGCAACCTATGAGATTCTG
4613 SEQ ID NO:3125 -14.9 -35.3 84.4 -20.4 0 -5.5
ATTGGCAACCTATGAGATTC
4615 SEQ ID NO:3126 -14.9 -33.1 81.2 -18.2 0 -4.9
AAACAAAAACATGTCCTCAT
4685 SEQ ID NO: 3127 -14.9 -287 75.6 -13.8 0 -6.2
CTCTGGCAGAGGAGCCGCTC
100 SEQ ID NO: 3128 -14.8 -44.8 95.6 -24.3 -4.5 -19.6
AAAATCAACCAAAAGTCTTC
308 SEQ ID NO: 3129 -14.8 -27.5 74.1 -12.7 0 -3.2
GAAAATCAACCAAAAGTCTT
309 SEQ ID NO: 3130 -14.8 -27.5 74.1 -12.7 0 -2.2
CTCTCCCATATACAGTCCCA
389 SEQ ID NO: 3131 -14.8 -38.1 88.2 -23.3 0 -07
CTGTCTCTCCCATATACAGT
393 SEQ ID NO:3132 -14.8 -35.8 85.1 -20.3 -0.4 -5.1
GTGGGGGCAGCAGACACAGC
568 SEQ ID NO:3133 -14.8 -45.1 96.9 -27.3 -1.8 -14
ACTCCTTCTCTGTGGGGGCA
579 SEQ ID NO:3134 -14.8 -43 94.8 -23.1 -5.1 -13
TACATCAGAGTGAGTTTTTG
602 SEQ ID NO: 3135 -14.8 -30.4 78.1 -15.1 -0.1 -3.2
TCATTCGTCTCTTTACCTGG
742 SEQ ID NO: 3136 -14.8 -34.5 83.6 -19.7 0 -3.1
CATTCGAGTTTCCTTCCAAA
840 SEQ ID NO:3137 -14.8 -31.6 79.7 -16.3 0 -7.6
TCATTCGAGTTTCCTTCCAA
841 SEQ ID NO: 3138 -14.8 -33.1 81.3 -17.8 0 -7.6
AATAGGCTTCTGATCCTGCT
1163 SEQ ID NO:3139 -14.8 -36.5 86.2 -20.1 -1.6 -9.8
TGCTGTTGAGGAGCTGGATG
1349 SEQ ID NO:3140 -14.8 -39.6 89.8 -22.9 -1.5 -117
ATTGTGCTGTCCTTCCACTG
1475 SEQ ID NO: 3141 -14.8 -37 87.1 -22.2 0 -3.8
TTTTATCGATGATGCAATCA
1515 SEQ ID NO: 3142 -14.8 -29 74.5 -12.5 -1.5 -11
CAGGTTCAATAACCTCCAAC
1737 SEQ ID NO:3143 -14.8 -33.7 83.4 -16.1 -2.8 -9.7
GAAACATACAGCATGTGTTT
2062 SEQ ID NO:3144 -14.8 -30.2 78.2 -12.5 -2.9 -10.4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular molecul Site oligo binding formation Duplex structure oligo oligo
GTTTGGAAGCAATAGTTAAG
2329 SEQ ID NO: 3145 -14.8 -29 77.5 -13.4 -0.6 -5.8
TGTTTGGAAGCAATAGTTAA
2330 SEQ ID NO: 3146 -14.8 -29 77.4 . -13.4 -0.6 -7.2
AAACTGATAGTTTATACAAT
2517 SEQ ID NO: 3147 -14.8 -25 70.3 -9.5 -0.3 -8.8
ACTGAAAAGTGATGACGACT
2650 SEQ ID NO: 3148 -14.8 -33.1 81.2 -16.5 -1.8 -5.5
TAGCCATTTTTGCCATATTA
3595 SEQ ID NO:3149 -14.8 -29.4 77.9 -14.6 0 -1.8
CAGTAGCTGAGCTTTCCTGT
3670 SEQ ID NO:3150 -14.8 -37.2 88.1 -19.3 -1.2 -14.3
AAATTTCTTGTGGCTTAGTA
3885 SEQ ID NO: 3151 -14.8 -29 77.5 -14.2 0 -5
TAGAAGTTCCATATTTTTAA
3961 SEQ ID NO: 3152 -14.8 -24.9 70.4 -8.5 -1.6 -6.2
ATATGATATATATCTGAAAC
4004 SEQ ID NO: 3153 -14.8 -25.2 69.1 -8.8 0 -11.4
CATTCTTATTTTACACTATA
4102 SEQ ID NO: 3154 -14.8 -24.1 69.2 -9.3 0 0
ACTGCACACAGGACCAGTCT
4286 SEQ ID NO: 3155 -14.8 -40.6 91.8 -24.2 -1.5 -10
GAGGATAACTTCCTCTGTAA
4336 SEQ ID NO: 3156 -14.8 -33.4 82.4 -11.7 -6.9 -16.2
GCAGAGGATAACTTCCTCTG
4339 SEQ ID NO: 3157 -14.8 -36.6 86.1 -11.7 -10.1 -22.6
CATTCAACTGCTGGGGGAGG
4529 SEQ ID NO: 3158 -14.8 -40.4 91.5 -24.1 -1.4 -9.1
AAATACTGAAAATAACAAAT
4728 SEQ ID NO: 3159 -14.8 -22.1 65.3 -7.3 0 -0.6
CAGATCCTTGGCACCTATTC
1224 SEQ ID NO: 3160 -14.7 -36.5 86 -21.8 0 -3.9
TGTTGTTGCTGTTGAGGAGC
1355 SEQ ID NO: 3161 -14.7 -36.9 87.4 -20.8 -1.3 -9.3
TCATTCTAATTTCATCAAAT
2187 SEQ ID NO: 3162 -14.7 -25 68.4 -10.3 0 0
GAAAAGTGATGACGACTCAA
2647 SEQ ID NO: 3163 -14.7 -32.1 79.5 -17.4 0 -7.2
ATACACCAACAGAAAGTCTA
2848 SEQ ID NO: 3164 -14.7 -31.8 80.8 -17.1 0 -5
TCTTAAACAGCTGTACAATA
3055 SEQ ID NO: 3165 -14.7 -29.5 78.1 -12.1 0 -13.6
TTAAGAAGTTATACAAACTA
3477 SEQ ID NO: 3166 -14.7 -25 71.4 -9.5 -0.6 -3.9
CTGTGTTACAGCTGGTTATC
3511 SEQ ID NO:3167 -14.7 -34 84 -17.7 -0.9 -11.2
GATGTCTGACATACAGTTCT
3703 SEQ ID NO:3168 -147 -33.4 81.1 -16.4 -0.3 -12.8
CCAAACTTCTGAAGCTTCTG
4553 SEQ ID NO:3169 -14.7 -33.3 82.6 -16.4 0 -12.5
GTGTATTATTGGCAACCTAT
4622 SEQ ID NO:3170 -14.7 -30.8 79.7 -16.1 0 -4.9 .
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GGAGCAGGAGGGAAATATAT
17 SEQ ID NO: 3171 -14.6 -35.8 85.2 -21.2 0 -2.7
CAAGCACACTGCTGGGGTTT
171 SEQ ID NO: 3172 -14.6 -38.7 907 -21.9 -2.2 -9.3
CCAGGTCATTTCCCATCACT
420 SEQ ID NO: 3173 -14.6 -37.3 86.9 -22 -0.4 -3.4
GTTGTTGCTGTTGAGGAGCT
1354 SEQ ID NO: 3174 -14.6 -36.9 87.4 -19.8 -2.5 -117
TTTACAGCTTCCACAAGTTA
1439 SEQ ID NO:3175 -14.6 -31.3 81.2 -15.8 -0.8 -5.4
TCTTTTGAAGAAAACTTTAC
1454 SEQ ID NO: 3176 -14.6 -25.3 71.2 -9.8 -0.8 -8.2
AATCATTCCTTCCAGCACAT
1500 SEQ ID NO: 3177 -14.6 -33.9 82.1 -19.3 0 -2.7
CAGAGCTATCATATCCTGCA
1767 SEQ ID NO: 3178 -14.6 -35.9 84.7 -20.4 -07 -5.2
CTGCAGTAGGGTCATTTGGT
1904 SEQ ID NO: 3179 -14.6 -37.1 88.1 -21.8 0 -8.9
TTAGGAACTGAAGAGAGAAG
2143 SEQ ID NO: 3180 -14.6 -33 817 -18.4 0 -5.5
TCTTATTAAGGCAGTCACTT
2461 SEQ ID NO: 3181 -14.6 -31.7 80.9 -17.1 0 -6.3
TTCACCATCTACTCTCCCAT
2671 SEQ ID NO: 3182 -14.6 -36.8 86.1 -22.2 0 0
TAAACAGCTGTACAATAACT
3052 SEQ ID NO: 3183 -14.6 -29.3 78.2 -12 0 -13.6
TCCTGAAAGACAAATAGTTT
3447 SEQ ID NO:3184 -14.6 -29.1 76.5 -14.5 0 -1.8
CTTCAAAAGGTCCTGAAAGA
3457 SEQ ID NO: 3185 -14.6 -32.3 81.1 -167 -0.7 -9.3
ATCAGGAAAGATTAACCAAT
3647 SEQ ID NO: 3186 -14.6 -29.4 76.2 -14.8 0 -3.6
GTACCATCAGGAAAGATTAA
3652 SEQ ID NO:3187 -14.6 -30.9 78.9 -15.4 -0.8 -4.2
GGGATGTGTAGTTTTACAAA
3727 SEQ ID NO:3188 -14.6 -30.3 79.3 -14.1 -1.6 -9
TATATCTGAAACTATAACTA
3996 SEQ ID NO:3189 -14.6 -26 71.9 -11.4 0 -1.7
GGACCAGTCTCCATCTCCCT
4276 SEQ ID NO:3190 -14.6 -42.5 92.6 -25.7 -2.2 -7.4
TTCACTGCACACAGGACCAG
4289 SEQ ID NO:3191 -14.6 -39.3 90.2 -22.9 -1.8 -6.7
GCAACCTTCACTGCACACAG
4295 SEQ ID NO: 3192 -14.6 -37.9 88.7 -21.2 -2.1 -5.9
TTAGATGGTGCCTTTAAGGA
4485 SEQ ID NO: 3193 -14.6 -34 84.3 -17.1 -0.7 -12.8
AAAATACTGAAAATAACAAA
4729 SEQ ID NO:3194 -14.6 -21.9 65.3 -7.3 0 -0.6
TCTCCCCTCTCCTGAGCAAG
187 SEQ ID NO:3195 -14.5 -42.2 92.8 -26.1 -1.6 -5.4
TGGAAAATCAACCAAAAGTC
311 SEQ ID NO:3196 -14.5 -29.9 77.6 -14.1 -1.2 -4.3 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
TTGGAAAATCAACCAAAAGT 312 SEQ ID NO:3197 -14.5 -28.4 75.8 -11.8 -2.1 -6.1
GGGAATCCCAGGTCATTTCC 427 SEQ ID NO: 3198 -14.5 -38.7 88.4 -20.4 -2.6 -15.6
TCTCCCGCCAGAGGAGAAAG 805 SEQ ID NO:3199 -14.5 -41.3 92 -21.2 -5.6 -14
TTAGAAAAAACTGTTCGACC 1285 SEQ ID NO: 3200 -14.5 -29.3 77.4 -14.8 0 -4
GTTTTGTTACCAGGATTTTC 1669 SEQ ID NO: 3201 -14.5 -28.9 76.9 -13.4 -0.9 -6
AGGAACTATTGTTTTGTTAC 1679 SEQ ID NO: 3202 -14.5 -27.9 75.8 -11.6 -1.8 -5.8
AGTCTGGAACAGAGCTATCA 1776 SEQ ID NO:3203 -14.5 -367 86.2 -20.5 -1.6 -10.8
CAGTAGGGTCATTTGGTCAT 1901 SEQ ID NO: 3204 -14.5 -35.1 84.6 -18.6 -2 -10.8
CCAGGAGTACTGCAGTAGGG 1913 SEQ ID NO: 3205 -14.5 -40.9 93.1 -22.5 -1.7 -16
TTAAGGCAACCATTCTTATT 2474 SEQ ID NO: 3206 -14.5 -29.1 77.3 -13.9 -0.4 -4.3
TTTCTTTAAGGCAACCATTC 2479 SEQ ID NO: 3207 -14.5 -30 78.5 -15.5 0 -5.4
TCGACTTTCTTTAAGGCAAC 2484 SEQ ID NO:3208 -14.5 -31.7 80.9 -17.2 0 -5.4
ACAGACTTTGGGCACTGGTG 2775 SEQ ID NO: 3209 -14.5 -39 90.5 -23.6 -0.6 -9.1
AGGTTTCCATGCATAAATCA 2898 SEQ ID NO.-3210 -14.5 -31.6 79.6 -15.5 -1 -10.9
ATACACTTGTAAACTTTTTT 2920 SEQ ID NO: 3211 -14.5 -24 69.7 -8.9 -0.3 -6.3
CCCATTTTCACACAGATGAT 3123 SEQ ID N0:3212 -14.5 -32.7 80.3 -16.9 -1.2 -4.9
TTTTTTTTTTTTTTTGACAA
3213 SEQ ID NO: 3213 -14.5 -18.2 58.5 -3.7 0 -0.5 TTTTTTTTTTTTTTTTGACA
3214 SEQ ID NO: 3214 -14.5 -18.2 58.5 -37 0 0 CAACCTATATAATTCTCCAC
3263 SEQ ID NO: 3215 -14.5 -30 77.8 -15.5 0 -3.3
CCATATAGCCATTGCAAAAA 3414 SEQ ID NO: 3216 -14.5 -30.2 78.8 -15 -0.5 -6.7
ATCTGGAATCACAACTTTTA 3494 SEQ ID NO: 3217 -14.5 -29.3 76.3 -14.8 0 -6.3
TGAATAGAAATCAAATTTCT 3897 SEQ ID NO: 3218 -14.5 -24.8 68.5 -9.4 -0.7 -6.5
CTTGAATAGAAATCAAATTT 3899 SEQ ID NO: 3219 -14.5 -23.3 66.7 -8.3 -0.2 -3.8
ATACCAATATGATATATATC 4010 SEQ ID NO: 3220 -14.5 -25.5 69.8 -10.3 0 -9
CTTCACTGCACACAGGACCA 4290 SEQ ID NO: 3221 -14.5 -39.3 90.2 -23 -1.8 -67
GCCAAACTTCTGAAGCTTCT 4554 SEQ ID NO: 3222 -14.5 -34.6 84.4 -17.9 0 -12.5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GGAATTAGTGTATTATTGGC
4629 SEQ ID NO: 3223 -14.5 -29.8 78 -15.3 0 -4.1
AAAAGCATTCAAAGAAAACA
4700 SEQ ID NO: 3224 -14.5 -26 71.9 -11.5 0 -2.7
GAATCCCAGGTCATTTCCCA
425 SEQ ID NO: 3225 -14.4 -37.5 87 -22.4 -0.4 -3.6
TGTTTCCCCCGAGGAAAGGC
464 SEQ ID NO: 3226 -14.4 -40.8 927 -23 -3.4 -11.2
TTCTTCCAAAAGCTTTAAGT
485 SEQ ID NO: 3227 -14.4 -28.6 76.9 -13.6 0 -8.6
GCTTGACAAAACCAGATCTC
914 SEQ ID NO: 3228 -14.4 -33.9 82.4 -19.5 0 -6.5
TTACTGGGGCTTGACAAAAC
922 SEQ ID NO: 3229 -14.4 -34.5 85.5 -20.1 0 -5.9
TGTATTCATGTCATAGTGGT
1127 SEQ ID NO: 3230 -14.4 -32.2 80.2 -17.1 -0.5 -5.3
GACATTAAAAATAGGCTTCT
1172 SEQ ID NO:3231 -14.4 -28.2 75.5 -13.8 0 -5
GTGGAATGACATTAAAAATA
1179 SEQ ID NO:3232 -14.4 -26 71.6 -11.1 -0.1 -5
CATTAGAAAAAACTGTTCGA
1287 SEQ ID NO: 3233 -14.4 -27 73.4 -12.6 0 -3.5
ATTCCTTTTATTTTTTTCTT
1612 SEQ ID NO: 3234 -14.4 -21.6 64.4 -7.2 0 0
CAGGAACTATTGTTTTGTTA
1680 SEQ ID NO: 3235 -14.4 -27.8 75.6 -11.6 -1.8 -5.8
GCCCATTTCACTGCTGCAAT
1846 SEQ ID NO: 3236 -14.4 -36.9 87 -22 0 -7.9
ACTCTCCCATCACTGAAAAG
2661 SEQ ID NO:3237 -14.4 -35 83.9 -20.6 0 -4.6
TACTCTCCCATCACTGAAAA
2662 SEQ ID NO: 3238 -14.4 -34.2 83.2 -19.8 0 -4.6
CCACATATTAAGGTTTCTAA
2717 SEQ ID NO: 3239 -14.4 -28.3 76.3 -13.9 0 -4
CTGTGAAAAAAAGTATGAAG
2809 SEQ ID NO: 3240 -14.4 -26.4 72.9 -12 0 -1
TACACCAACAGAAAGTCTAG
2847 SEQ ID NO: 3241 -14.4 -32.8 82.6 -18.4 0 -5
CTTGAATAAAAAATTATGTA
3034 SEQ ID NO: 3242 -14.4 -21.2 64.1 -6.8 0 -0.9
AGACAAATAGTTTACCAGCT
3440 SEQ ID NO: 3243 -14.4 -31.5 81 -16.6 -0.2 -3.5
AAAGGTCCTGAAAGACAAAT
3452 SEQ ID NO: 3244 -14.4 -31.1 79.4 -14.8 -1.9 -9
TCAGGAAAGATTAACCAATT
3646 SEQ ID NO:3245 -14.4 -29.2 76.4 -14.8 0 -3.6
CTTTCCTGTACCATCAGGAA
3659 SEQ ID NO: 3246 -14.4 -35.1 84.8 -14.1 -6.6 -15.6
TCAGTAGCTGAGCTTTCCTG
3671 SEQ ID NO: 3247 -14.4 -37.4 87.9 -20.4 -1.2 -13.3
CCCTAGAGCAAACTGTTTGG
4252 SEQ ID NO: 3248 -14.4 -35.6 86.8 -19.7 -0.9 -10.6
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
ATCTCCCTCTTCCCCTAGAG
4264 SEQ ID NO:3249 -14.4 -40.3 90.6 -23.6 -2.3 -7
TAAGCACCACCTTCCTGTCT
4463 SEQ ID NO:3250 -14.4 -38.4 89.9 -24 0 -2.7
TGGTGCCTTTAAGGATGTAA
4480 SEQ ID NO: 3251 -14.4 -33.8 84.4 -18.4 -0.5 -9.8
AAACATGTCCTCATTTTATT
4679 SEQ ID NO: 3252 -14.4 -26.8 72.5 -12.4 0 -6.3
TCTCCTGAGCAAGCACACTG
180 SEQ ID NO: 3253 -14.3 -39.5 90 -25.2 0 -5.7
TTGGACAGATCTGGCTGCTG
352 SEQ ID NO: 3254 -14.3 -39.6 89.8 -22.1 -2.2 -14.4
CAGAAGAAAACTCCAAATCC
717 SEQ ID NO: 3255 -14.3 -31.3 79.2 -17 0 -0.3
AGGTACTCACACCATGAACA
1092 SEQ ID NO:3256 -14.3 -36.3 86.4 -19.7 -2.3 -6.4
CTTCTGATCCTGCTGTTGAG
1157 SEQ ID NO:3257 -14.3 -36.2 85.1 -21.9 0 -3.4
AGGTAATTGTGCTGTCCTTC
1480 SEQ ID NO:3258 -14.3 -35 85 -19.8 -0.8 -7
TGATTGGTGATGATTTCAGC
2392 SEQ ID NO: 3259 -14.3 -33 80.2 -17.6 -1 -5.7
TTTAAGGCAACCATTCTTAT
2475 SEQ ID NO: 3260 -14.3 -29.1 77.3 -14.1 -0.4 -4.3
ACAATAAAAAATAAAACAAC
2553 SEQ ID NO: 3261 -14.3 -20.5 63.5 -6.2 0 0
TGCGTATTTAAAACAAAACA
2583 SEQ ID NO: 3262 -14.3 -25.5 72.3 -11.2 0 -5.5
CCATCCTTCTTTGACTGTGG
2746 SEQ ID NO: 3263 -14.3 -35.9 85.5 -21 -0.3 -6.3
TCCTAACTATACAGGGGGGG
2869 SEQ ID NO: 3264 -14.3 -40.1 92.8 -21.2 -4.6 -13.6
ACACTTGTAAACTTTTTTTC
2918 SEQ ID NO:3265 -14.3 -24.9 71.3 -10.6 0 -2.7
ATAGCTATAAAAGGCACAAC
2954 SEQ ID NO: 3266 -14.3 -30.6 79.9 -13.8 -2.5 -8.7
ATAAAAAATTATGTAAGAAA
3029 SEQ ID NO: 3267 -14.3 -20 61.6 -5.7 0 -4.2
AGCTGCCCATCTTAAACAGC
3064 SEQ ID NO: 3268 -14.3 -37.1 88.1 -20.5 -2.3 -8.1
CGAACTAGCTGCCCATCTTA
3070 SEQ ID NO: 3269 -14.3 -36.8 87.6 -22.5 0 -6.2
ATAGTTTACCAGCTTTCTTG
3434 SEQ ID NO:3270 -14.3 -30.2 79.2 -15.9 0 -4
GTCCTGAAAGACAAATAGTT
3448 SEQ ID NO:3271 -14.3 -30.4 78.4 -14.2 -1.9 -5.8
GTTTTACAAACATGGATGTC
3717 SEQ ID NO: 3272 -14.3 -29.2 76.6 -14 0 -9.7
TTCAACAGTGAAGAAATTCA
3780 SEQ ID NO: 3273 -14.3 -29 75.5 -12.1 -2.6 -6.7
GTAGCCCTTCCCTTCCCAGA
4037 SEQ ID NO: 3274 -14.3 -42.1 94.2 -27.8 0 -1.3 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CTTCTGTTGTCATTCAACTG
4539 SEQ ID NO: 3275 -14.3 -31.2 79 -14.6 -2.3 -7.6
ATTATTGGCAACCTATGAGA
4618 SEQ ID NO: 3276 -14.3 -32 80.4 -17 -0.5 -6.5
AGCCAGTGAGGGTGAAGACG
260 SEQ ID NO: 3277 -14.2 -41 92.1 -24.5 -2.3 -8.6
TGCTTTCTTCCAAAAGCTTT
489 SEQ ID NO: 3278 -14.2 -30.6 79.6 -13.1 -3.3 -8.8
GGGGCAGCAGACACAGCAGT
565 SEQ ID NO: 3279 -14.2 -43.9 95.9 -287 -0.9 -4.7
AATCCTGCAAAATGTCAAAG
702 SEQ ID NO: 3280 -14.2 -29.8 77.3 -15.6 0 -4.8
CCAAATCCTGCAAAATGTCA
705 SEQ ID NO:3281 -14.2 -32.2 80.7 -18 0 -4.9
AGAGTCCCCAGAGAAGTCAA
1255 SEQ ID NO: 3282 -14.2 -38.7 88.8 -24.5 0 -3.8
AGCTTCCACAAGTTAAGACT
1434 SEQ ID NO: 3283 -14.2 -33.6 83.7 -18.5 -0.8 -4.8
TTGTGCTGTCCTTCCACTGC
1474 SEQ ID NO: 3284 -14.2 -39.3 90.3 -25.1 0 -3.8
AAACATCCAGGAGTACTGCA
1919 SEQ ID NO: 3285 -14.2 -36.2 86.5 -20.3 -1.7 -8.7
TGGAAGCAATAGTTAAGGAG
2326 SEQ ID NO: 3286 -14.2 -32.8 82.5 -18.6 0 -2.5
CAACCATTCTTATTAAGGCA
2468 SEQ ID NO: 3287 -14.2 -30.3 79 -16.1 0 -7.2
ATTCGACTTTCTTTAAGGCA
2486 SEQ ID NO: 3288 -14.2 -30.6 78.9 -16.4 0 -5.4
CAATAAAAAATAAAACAACA
2552 SEQ ID NO: 3289 -14.2 -20.4 63.2 -6.2 0 0
GGATACACCAACAGAAAGTC
2850 SEQ ID NO:3290 -14.2 -34.1 83.2 -19.2 -0.5 -3.4
AAGGCACAACTTCCCTTTTC
2944 SEQ ID NO:3291 -14.2 -34.3 84.7 -19.6 -0.2 -4.6
AATAGCTATAAAAGGCACAA
2955 SEQ ID NO: 3292 -14.2 -29.3 78.1 -12.6 -2.5 -9.8
CATCAGGTTAGAAGCACCAA
3143 SEQ ID NO: 3293 -14.2 -34.9 84.8 -19.2 -1.4 -5.8
GATATATACAAAATATGAGC
3236 SEQ ID NO: 3294 -14.2 -26 71.3 -11.8 0 -3.5
CTCTCAGCTGTGTTACAGCT
3518 SEQ ID NO: 3295 -14.2 -37.3 87.9 -17.3 -5.5 -19.3
TGGGAATGTGAAAATGGGTG
3617 SEQ ID NO: 3296 -14.2 -34.6 84 -20.4 0 0
TTTTCAACAGTGAAGAAATT
3782 SEQ ID NO:3297 -14.2 -26.3 71.9 -9.8 -2.3 -9.6
TTAGTGCAAGGGGAGATTGA
4420 SEQ ID NO: 3298 -14.2 -36.3 86.5 -22.1 0 -4.8
AATTAGTGTATTATTGGCAA
4627 SEQ ID NO: 3299 -14.2 -27.1 74.2 -12.9 0 -4.3
GAAACCTTCACAGTAGCTCC
238 SEQ ID NO: 3300 -14.1 -36.3 86.6 -22.2 0 -4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GTTTTTGGAAACTCCTTCTC
589 SEQ ID NO: 3301 -14.1 -31.2 79.6 -15.2 -1 -11.8
TTCAAATGTTGCTGTTCTGA
625 SEQ ID NO: 3302 -14.1 -31.2 79.1 -17.1 0 -1.8
GAGGAGAAAGCAAACAGTTT
795 SEQ ID NO: 3303 -14.1 -32.2 81.2 -18.1 0 -2.7
CTTTTGAAGAAAACTTTACA
1453 SEQ ID NO: 3304 -14.1 -25 71.1 -9.8 -1 -5.1
TCAGAGGAAACATACAGCAT
2068 SEQ ID NO: 3305 -14.1 -34.2 82.9 -20.1 0 -27
GCTCTTGGCTCTTCAGACCG
2166 SEQ ID NO: 3306 -14.1 -40 90.4 -24 -1.9 -8.3
CAGAAGTTTTTTGATATTTC
2432 SEQ ID NO: 3307 -14.1 -24.4 69.1 -9.4 -07 -7.4
TAAGGCAACCATTCTTATTA
2473 SEQ ID NO: 3308 -14.1 -29.5 78.1 -14.9 -0.2 -3.9
AACAGAAAGTCTAGAAAATT
2841 SEQ ID NO: 3309 -14.1 -26.7 72.8 -12.1 0 -7.5
CATCTTAAACAGCTGTACAA
3057 SEQ ID NO:3310 -14.1 -30.3 79 -13.5 0 -13.6
CAATTTTTGTGGTCTTCTGA
3174 SEQ ID NO:3311 -14.1 -29.9 77.4 -15.8 0 -2.4
TCAATTTTTGTGGTCTTCTG
3175 SEQ ID NO: 3312 -14.1 -29.9 77.4 -15.8 0 -2.4
CAGGAAAGATTAACCAATTG
3645 SEQ ID NO:3313 -14.1 -28.9 76.4 -14.8 0 -3.6
TTGAATAGAAATCAAATTTC
3898 SEQ ID NO: 3314 -14.1 -23.6 66.8 -9 -0.1 -3.5
ACCCCTACAAAAAATATATA
4155 SEQ ID NO:3315 -14.1 -27.4 75.2 -13.3 0 -3.3
TATGTGAAAGTAACCCGCTA
4505 SEQ ID NO:3316 -14.1 -33.4 84 -18.8 -0.2 -3.4
TGAAGCTTCTGTTGTCATTC
4544 SEQ ID NO: 3317 -14.1 -32.7 807 -16.4 -1.2 -12.5
TTCAAAGAAAACAAAAACAT
4693 SEQ ID NO: 3318 -14.1 -23.6 67.9 -9.5 0 -1.4
AAAAAGCATTCAAAGAAAAC
4701 SEQ ID NO:3319 -14.1 -24.8 70.2 -10.7 0 -2.5
TCCTGAGCAAGCACACTGCT
178 SEQ ID NO: 3320 -14 -40.5 91.8 -24.8 -1.7 -7.3
TGAGAGTGAAACTGCTTTGG
368 SEQ ID NO:3321 -14 -34.6 84.2 -18.2 -2.4 -7.6
TCTTCCAAAAGCTTTAAGTC
484 SEQ ID NO: 3322 -14 -30.1 78.6 -15.5 0 -8.6
AACTCCAAATCCTGCAAAAT
709 SEQ ID NO: 3323 -14 -31 79.2 -17 0 -4.9
ATTCGTCTCTTTACCTGGGG
740 SEQ ID NO: 3324 -14 -36.6 87 -21.2 -1.2 -10.1
CTCATTCGTCTCTTTACCTG
743 SEQ ID NO: 3325 -14 -33.3 81.9 -19.3 0 0
ACTCTCATTCGTCTCTTTAC
746 SEQ ID NO: 3326 -14 -32.5 80.3 -18.5 0 0 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
ATTTTATCGATGATGCAATC
1516 SEQ ID NO: 3327 -14 -28 72.8 -12.5 0 -11
GGATTTTCAGAGGTTTCTTG
1657 SEQ ID NO: 3328 -14 -31.3 79.2 -16.8 0 -7.6
GGTCATTTGGTCATCCAGGT
1895 SEQ ID NO: 3329 -14 -37.4 87 -19.8 -3.6 -11.1
CTTAGGAACTGAAGAGAGAA
2144 SEQ ID NO:3330 -14 -33 81.7 -19 0 -6.3
ATGAAAACAATAAAAAATAA
2559 SEQ ID NO:3331 -14 -19.6 60.7 -5.6 0 -0.4
CACCATCTACTCTCCCATCA
2669 SEQ ID NO:3332 -14 -38 87.3 -24 0 0
AACTTGAATAAAAAATTATG
3036 SEQ ID NO:3333 -14 -20.8 63.3 -6.8 0 -0.9
CAATAACTTGAATAAAAAAT
3040 SEQ ID NO: 3334 -14 -20.8 63.3 -6.8 0 -0.8
ATCTTAAACAGCTGTACAAT
3056 SEQ ID NO:3335 -14 -29.3 77.2 -12.6 0 -13.6
CAAAAGGTCCTGAAAGACAA
3454 SEQ ID NO:3336 -14 -32.1 81.2 -16.2 -1.9 -9
GAAATCAAATTTCTTGTGGC
3891 SEQ ID NO:3337 -14 -28.9 75.8 -14.3 -0.3 -5.5
CCCCTACAAAAAATATATAA
4154 SEQ ID NO:3338 -14 -26.1 73.3 -12.1 0 -3.4
ATCATATAGGTTATCCATCA
4186 SEQ ID NO:3339 -14 -30.6 77.2 -15.4 -1.1 -5.2
CAGTCTCCATCTCCCTCTTC
4272 SEQ ID NO:3340 -14 -39.1 88 -25.1 0 -1.3
AGCAACCTTCACTGCACACA
4296 SEQ ID NO:3341 -14 -37.9 89.1 -21.6 -2.3 -5.9
GATAACTTCCTCTGTAATCT
4333 SEQ ID NO: 3342 -14 -31.2 78.8 -15.8 -1.3 -4.8
AATACTGAAAATAACAAATA
4727 SEQ ID NO: 3343 -14 -22.5 66.1 -8.5 0 -0.6
ATCACATCTCCCCTCTCCTG
193 SEQ ID NO: 3344 -13.9 -40.2 89.4 -26.3 0 0
CTTTGGACAGATCTGGCTGC
354 SEQ ID NO:3345 -13.9 -38.4 88.3 -21.4 -1 -14.4
TTTCTCTTGCTTAATTACCC
1001 SEQ ID NO: 3346 -13.9 -30.4 79.3 -16.5 0 -2.3
CTCTTTTGAAGAAAACTTTA
1455 SEQ ID NO: 3347 -13.9 -25.2 71.1 -10.4 -0.5 -9
TTATCGATGATGCAATCATT
1513 SEQ ID NO:3348 -13.9 -29.2 74.3 -13.6 -1.3 -11-.2
GAATTTTATCGATGATGCAA
1518 SEQ ID NO:3349 -13.9 -27.8 72.9 -12.5 0 -10.8
TGGTAACGTTGCAGGAACTA
1691 SEQ ID NO:3350 -13.9 -35.3 86.7 -18.2 0 -14.6
ATATCCTGCATATAACACTT
1757 SEQ ID NO:3351 -13.9 -30 77.6 -16.1 0 -4.5
CTGCTGCAATCACTTGCCGC
1836 SEQ ID NO: 3352 -13.9 -39.5 90.3 -21.5 -4.1 -11 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
AATGCCATAAGAAACATCCA 1930 SEQ ID NO:3353 -13.9 -31.6 79.6 -17.7 0 -3.4
GCACTTGATTGTCTATATGA 1966 SEQ ID NO: 3354 -13.9 -31.2 78.5 -17.3 0 -4.2
TTTCATCAAATAGCTCTTGG 2178 SEQ ID NO: 3355 -13.9 -30.4 78 -16.5 0 -4.8
ACAATGGCTTTTCCTAGCTC 2218 SEQ ID NO: 3356 -13.9 -34.9 85 -18.4 -2.6 -8.5
TTATCCAAAAATGTTTGGAA 2341 SEQ ID NO: 3357 -13.9 -26.5 72.6 -7.7 -4.9 -15
GATTGGTGATGATTTCAGCT 2391 SEQ ID NO: 3358 -13.9 -33 80.3 -18.4 -0.5 -5.7
CAACAGAAAGTCTAGAAAAT 2842 SEQ ID NO: 3359 -13.9 -27.9 74.7 -13.5 0 -7.5
TCCATGCATAAATCAAAAAT 2893 SEQ ID NO: 3360 -13.9 -26.9 72.3 -13 0 -7
ACAGCTGGTTATCTGGAATC 3504 SEQ ID NO:336i -13.9 -35.4 84.5 -19.8 -1.4 -11.1
TCTAGCCATTTTTGCCATAT 3597 SEQ ID NO: 3362 -13.9 -317 80.5 -17.8 0 -2.4
GGTGTCTAGCCATTTTTGCC 3601 SEQ ID NO: 3363 -13.9 -35.9 86.6 -19.3 -2.7 -9.2
CAGATGGGAATGTGAAAATG 3621 SEQ ID NO:3364 -13.9 -31.4 78.7 -16.8 -0.5 -3.8
GAATGCATGTACAAACTATA
4207 SEQ ID NO: 3365 -13.9 -28.7 76 -14.1 0 -9.1 TGAATGCATGTACAAACTAT
4208 SEQ ID NO: 3366 -13.9 -29.5 76.9 -14.9 0 -9.1 ATGAATGCATGTACAAACTA
4209 SEQ ID NO: 3367 -13.9 -29.5 76.9 -14.9 0 -8.8 TTAAATAATTTCTCCAAAAT
4744 SEQ ID NO: 3368 -13.9 -22.2 65.2 -8.3 0 -0.3 CCTGCTGTGGGAATCCCAGG
435 SEQ ID NO: 3369 -13.8 -42.8 94.2 -24.1 -4.1 -177 CCCTGCTGTGGGAATCCCAG
436 SEQ ID NO: 3370 -13.8 -42.8 94.2 -24.1 -4.3 -17.7 CAGTGGATGCTGAACTCTTG
549 SEQ ID NO: 3371 -13.8 -36 85.2 -19.3 -2.9 -10.6
ACAAAACCAGATCTCCATTA 909 SEQ ID NO: 3372 -13.8 -31.7 79.8 -17.9 0 -6.5
TTTATTACCAATTATATTTG 1058 SEQ ID NO: 3373 -13.8 -21.5 64.9 -7.7 0 -3.8
TTTTGAAGAAAACTTTACAG 1452 SEQ ID NO: 3374 -13.8 -25 71.1 -10.1 -1 -5.1
CAATCATTCCTTCCAGCACA 1501 SEQ ID NO: 3375 -13.8 -34.9 83.7 -21.1 0 -2.7
CCTTTTATTTTTTTCTTTGT 1609 SEQ ID NO: 3376 -13.8 -22.4 66.9 -8.6 0 0
ACTGATAGTTTATACAATAA 2515 SEQ ID NO: 3377 -13.8 -25.4 71.1 -11.6 0 -4
TAAATCAAAAATGCTATCCT 2885 SEQ ID NO: 3378 -13.8 -27.1 73.3 -13.3 0 -1.7 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CTTTTCTGATATACACTTGT
2930 SEQ ID NO: 3379 -13.8 -28.3 75.3 -14.5 0 -1.7
AAAAGGCACAACTTCCCTTT
2946 SEQ ID NO:3380 -13.8 -32.8 83.3 -17.6 -1.3 -6.8
AGCTATAAAAGGCACAACTT
2952 SEQ ID NO:3381 -13.8 -31.2 81.1 -14.9 -2.5 -6.2
TGTGTTACAGCTGGTTATCT
3510 SEQ ID NO:3382 -13.8 -34 84.2 -18.6 0 -11.2
TTCCTGTACCATCAGGAAAG
3657 SEQ ID NO: 3383 -13.8 -35.1 84.8 -14.1 -7.2 -16.8
TGTAGTTTTACAAACATGGA
3721 SEQ ID NO: 3384 -13.8 -29.1 77.4 -14.7 -0.1 -8.3
ATTAGAAGTTCCATATTTTT
3963 SEQ ID NO:3385 -13.8 -247 69.5 -10.9 0 -4.9
TTTTAATAAATTGCATGTAA
4064 SEQ ID NO:3386 -13.8 -22.2 66 -7.5 0 -9.6
GCAAGGGGAGATTGAGTAAA
4415 SEQ ID NO:3387 -13.8 -35.1 85 -21.3 0 -2.7
TGCAAGGGGAGATTGAGTAA
4416 SEQ ID NO: 3388 -13.8 -36.3 86.5 -22.5 0 -4.1
ATGGTGCCTTTAAGGATGTA
4481 SEQ ID NO: 3389 -13.8 -34 84.1 -19.1 -0.7 -9.8
GAAGCTTCTGTTGTCATTCA
4543 SEQ ID NO: 3390 -13.8 -32.7 80.7 -16.7 -0.7 -12.5
CAAATCCTGCAAAATGTCAA
704 SEQ ID NO:3391 -13.7 -29.8 77.3 -16.1 0 -4.9
CATTCGTCTCTTTACCTGGG
741 SEQ ID NO:3392 -137 -35.4 85.2 -21.2 0 -7.7
TCTGATCTCCAAGGACTCTC
760 SEQ ID NO:3393 -13.7 -38.2 86.6 -24.5 0 -6.9
CAAACAGTTTTCATCTATCA
785 SEQ ID NO: 3394 -137 -28.1 74.4 -14.4 0 -3.3
CTGTTGTTGCTGTTGAGGAG
1356 SEQ ID NO: 3395 -13.7 -35.6 85.7 -21.9 0 -2.9
GTCATTTGGTCATCCAGGTG
1894 SEQ ID NO:3396 -13.7 -36.2 85.2 -16.5 -6 -15.2
TTCTAATTTCATCAAATAGC
2184 SEQ ID NO:3397 -13.7 -26.2 71.5 -12.5 0 -0.6
TCTGATTGGTGATGATTTCA
2394 SEQ ID NO:3398 -137 -32 78.5 -18.3 0 -5.3
ATTTTGGTATCTGATTGGTG
2403 SEQ ID NO:3399 -13.7 -30.4 77.9 -16.7 0 -5.4
TCACCATCTACTCTCCCATC
2670 SEQ ID NO:3400 -137 -38.3 87.2 -24.6 0 0
AAATTTCACCATCTACTCTC
2675 SEQ ID NO: 3401 -13.7 -30.8 78.1 -17.1 0 0
ACGTCCACATATTAAGGTTT
2721 SEQ ID NO: 3402 -13.7 -30.8 79.6 -17.1 0 -2.7
TGTGAAAAAAAGTATGAAGA
2808 SEQ ID NO: 3403 -13.7 -26.7 72.7 -13 0 -1
GCAAGCGTAGTTCACTAAAT
3003 SEQ ID NO: 3404 -13.7 -31.9 81.4 -177 -0.2 -5.6 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TAAAAAATTATGTAAGAAAA
3028 SEQ ID NO: 3405 -13.7 -19.8 61.6 -6.1 0 -4.2
TTAAACAGCTGTACAATAAC
3053 SEQ ID NO: 3406 -13.7 -28.1 76.5 -11.7 0 -13.6
TGGTCTTCTGATAGCTAAGT
3165 SEQ ID NO: 3407 -13.7 -34.4 84 -19.6 -0.8 -9.7
TTTTTGTGGTCTTCTGATAG
3171 SEQ ID NO: 3408 -13.7 -30.3 78.2 -15.7 -0.8 -4.3
AATTTTTGTGGTCTTCTGAT
3173 SEQ ID NO: 3409 -137 -28.9 75.5 -14.7 -0.1 -2.4
GATTTGAGTCAATTTTTGTG
3183 SEQ ID NO: 3410 -13.7 -26.5 72 -11.3 -0.5 -11
TAGTTATTTTTTAAATTAGT
3363 SEQ ID NO: 3411 -13.7 -20 62.6 -6.3 0 -1.9
AATAGTTTACCAGCTTTCTT
3435 SEQ ID NO: 3412 -137 -29 77.5 -15.3 0 -4
GACAAATAGTTTACCAGCTT
3439 SEQ ID NO: 3413 -13.7 -30.3 79.3 -16.6 0 -4
AGGTGCACACAGAAAGGGCT
3830 SEQ ID NO: 3414 -13.7 -40.2 92.3 -25 0 -11
CAGAAAGTTGGTAAGGTGCA
3843 SEQ ID NO: 3415 -13.7 -34.5 85.3 -20.8 0 -4.8
TGCACACAGGACCAGTCTCC
4284 SEQ ID NO: 3416 -137 -42 92.9 -27.4 -0.7 -8.9
GCTTTAAGTCTGTTTCCCCC
474 SEQ ID NO:3417 -13.6 -35.9 87 -22.3 0 -4.3
TGCAATGCTTTCTTCCAAAA
494 SEQ ID NO:3418 -13.6 -30.8 79.3 -16.7 0 -7.6
TCTCATTCGTCTCTTTACCT
744 SEQ ID NO: 3419 -13.6 -33.6 82 -20 0 0
GTTTCCTTCCAAAAGGAATG
833 SEQ ID NO:3420 -13.6 -31 79.4 -12.9 -4.5 -12.5
ATTAGAAAAAACTGTTCGAC
1286 SEQ ID NO:3421 -13.6 -27.1 73.6 -13.5 0 -4
TAGCCATTAGAAAAAACTGT
1291 SEQ ID NO: 3422 -13.6 -29 77.5 -15.4 0 -1.4
TGCTGAATTCCTTTTATTTT
1618 SEQ ID NO: 3423 -13.6 -26.5 72.6 -12.9 0 -5.2
CTCCAAGTTGAGTCTGGAAC
1786 SEQ ID NO: 3424 -13.6 -36.1 85.6 -19.4 -2.6 -14.1
CAATGGCTTTTCCTAGCTCT
2217 SEQ ID NO:3425 -13.6 -34.8 85 -18.4 -2.8 -8.7
TAAAACAACAAAACCTCTAC
2542 SEQ ID NO:3426 -13.6 -27.7 76 -14.1 0 0
AATTTCTGGGATATATTAAC
2699 SEQ ID NO: 3427 -13.6 -26.5 72.4 -12.9 0 -6.8
CGTAGTTCACTAAATATAAA
2998 SEQ ID NO: 3428 -13.6 -25.5 72 -11.4 -0.2 -5.6
GGAAAGCTACGAACTAGCTG
3079 SEQ ID NO:3429 -13.6 -35.5 86.3 -17.9 -4 -11.4
CTTTTAAGAAGTTATACAAA
3480 SEQ ID NO:3430 -13.6 -23.3 68.6 -9.7 0 -5.2 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
AGCTGTGTTACAGCTGGTTA
3513 SEQ ID NO: 3431 -13.6 -36 87.7 -16 -6.2 -20.4
CTGAAACTATAACTAAATAT
3991 SEQ ID NO: 3432 -13.6 -24.1 69.3 -10.5 0 -1
CTGCCTGTATGAATGCATGT
4217 SEQ ID NO: 3433 -13.6 -35.1 84.2 -19.9 -1.4 -10.8
TCCTGTTTACATACTTTACA
4444 SEQ ID NO: 3434 -13.6 -29.5 78.2 -15.9 0 -4.4
TCTCCTGTTTACATACTTTA
4446 SEQ ID NO:3435 -13.6 -297 78.2 -16.1 0 -4.3
CTATTGCCAAACTTCTGAAG
4559 SEQ ID NO: 3436 -13.6 -31.2 79.9 -17.6 0 -7.3
CTATGAGATTCTGCACTATT
4606 SEQ ID NO:3437 -13.6 -31.1 78.5 -17.5 0 -6
CAAAAACATGTCCTCATTTT
4682 SEQ ID NO: 3438 -13.6 -27.4 73.7 -13.8 0 -5.9
AAAGCATTCAAAGAAAACAA
4699 SEQ ID NO:3439 -13.6 -26 71.9 -12.4 0 -2.7
GAGTGAAACTGCTTTGGACA
365 SEQ ID NO: 3440 -13.5 -347 84.3 -20.3 -0.8 -7.6
AGAGTGAAACTGCTTTGGAC
366 SEQ ID NO:3441 -13.5 -34.7 84.3 -19.2 -2 -5.2
TTTGGAAACTCCTTCTCTGT
586 SEQ ID NO: 3442 -13.5 -33.6 82.8 -19 -1 -6.3
CTCCAAATCCTGCAAAATGT
707 SEQ ID NO: 3443 -13.5 -32.2 80.7 -18.7 0 -4.9
TTCGTCTCTTTACCTGGGGA
739 SEQ ID NO: 3444 -13.5 -37.9 88.9 -21.2 -3.2 -10.1
CTCTCATTCGTCTCTTTACC
745 SEQ ID NO: 3445 -13.5 -33.6 81.9 -20.1 0 0
GCTTCCACAAGTTAAGACTC
1433 SEQ ID NO: 3446 -13.5 -33.9 83.5 -20.4 0 -4.9
TCGAATTTTATCGATGATGC
1520 SEQ ID NO: 3447 -13.5 -29.6 75.2 -13.9 -2.2 -11
ACCAGGATTTTCAGAGGTTT
1661 SEQ ID NO: 3448 -13.5 -33.5 82.8 -19.3 -0.4 -6.5
GTGGTAACGTTGCAGGAACT
1692 SEQ ID NO:3449 -13.5 -36.2 87.4 -19.5 -0.4 -14.6
CTATCATATCCTGCATATAA
1762 SEQ ID NO: 3450 -13.5 -29.5 76.3 -16 0 -4.5
GTTTGCACTTGATTGTCTAT
1970 SEQ ID NO: 3451 -13.5 -30.4 78.1 -16.9 0 -4.9
TTGCCAAGTCTTGGCCCTCT
2618 SEQ ID NO: 3452 -13.5 -40.3 92.3 -20 -4.3 -21.8
CCAGCCAACTGTGAAAAAAA
2817 SEQ ID NO: 3453 -13.5 -31.6 81.3 -17.3 -0.6 -3.9
TTAATGTTTAGAGTTTATTT
3100 SEQ ID NO: 3454 -13.5 -22.3 66.2 -8.8 0 -3.8
TAAGTGCCATCAGGTTAGAA
3150 SEQ ID NO: 3455 -13.5 -34.1 84.2 -19.2 -1.3 -5.3
ATTTTTGTGGTCTTCTGATA
3172 SEQ ID NO: 3456 -13.5 -29.3 76.3 -14.9 -0.8 -4.3 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TAGCCATTGCAAAAATAGGG
3409 SEQ ID NO: 3457 -13.5 -32.4 82.5 -17.9 -0.9 -6.7
AAGGTCCTGAAAGACAAATA
3451 SEQ ID NO: 3458 -13.5 -31.5 80.1 -16.1 -1.9 -9
TTTAGTGAGAGGAATTACTT
3553 SEQ ID NO: 3459 -13.5 -29.5 77.3 -13.9 -2.1 -7.5
CATGGATGTCTGACATACAG
3707 SEQ ID NO: 3460 -13.5 -34.4 82.3 -17.7 -1.1 -14.5
AAAAAGCAAATGATTGTTTT
4768 SEQ ID NO: 3461 -13.5 -23.5 67.8 -7.6 -2.4 -7.6
GAGCAGGAGGGAAATATATT
16 SEQ ID NO: 3462 -13.4 -33.4 82.1 -20 0 -3.2
AAACAACTTAGCTTGTGAAC
43 SEQ ID NO: 3463 -13.4 -29.8 787 -157 -0.5 -7.1
AGTTTTTGGAAACTCCTTCT
590 SEQ ID NO: 3464 -13.4 -30.9 797 -15.2 -1 -12.8
CGCCAGAGGAGAAAGCAAAC
800 SEQ ID NO: 3465 -13.4 -37.3 87.8 -23.9 0 -3.1
TGATCCTGCTGTTGAGAAAG
1153 SEQ ID NO: 3466 -13.4 -35 83.8 -21.6 0 -3.4
AATTCCTTTTATTTTTTTCT
1613 SEQ ID NO: 3467 -13.4 -21.6 64.4 -8.2 0 0
CTTTCTTTAAGGCAACCATT
2480 SEQ ID NO: 3468 -13.4 -29.7 78.5 -16.3 0 -5.4
ATAAATTTCACCATCTACTC
2677 SEQ ID NO: 3469 -13.4 -28.7 75.3 -15.3 0 -1.1
ATCACACAGACTTTGGGCAC
2780 SEQ ID NO: 3470 -13.4 -37.1 87.2 -23.7 0 -5.6
AAAATAATTTTCAACAGTGA
3789 SEQ ID NO: 3471 -13.4 -24.2 68.6 -10.8 0 -3.2
TAAAAATATATTAGAAGTTC
3972 SEQ ID NO: 3472 -13.4 -21.9 65.3 -8.5 0 -5.1
ACAATCATTTTAATAAATTG
4071 SEQ ID NO: 3473 -13.4 -21 63.2 -7.6 0 -2.3
TATGAATGCATGTACAAACT
4210 SEQ ID NO: 3474 -13.4 -29.5 76.9 -15.4 0 -9.1
GCCTGTATGAATGCATGTAC
4215 SEQ ID NO: 3475 -13.4 -34.4 83.5 -18.9 -1.9 -11.8
CCTTCACTGCACACAGGACC
4291 SEQ ID NO: 3476 -13.4 -40.5 91.5 -25.3 -1.8 -8.3
CTCCTGTTTACATACTTTAC
4445 SEQ ID NO: 3477 -13.4 -29.5 78.3 -16.1 0 -4.4
GTCTCCTGTTTACATACTTT
4447 SEQ ID NO: 3478 -13.4 -30.6 79.2 -17.2 0 -4.4
ATACTGAAAATAACAAATAA
4726 SEQ ID NO:3479 -13.4 -22.5 66.1 -9.1 0 -0.6
GTTAATGATTCTTTGGAGTC
136 SEQ ID NO:3480 -13.3 -29.6 76.5 -13.3 -3 -12.6
GCACACTGCTGGGGTTTTCT
168 SEQ ID NO: 3481 -13.3 -39 907 -24.7 -0.9 -6.1
GAGAGTGAAACTGCTTTGGA
367 SEQ ID NO: 3482 -13.3 -34.9 84.2 -19.2 -2.4 -6.6 . .
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
TCCTTCTCTGTGGGGGCAGC
577 SEQ ID NO: 3483 -13.3 -44.2 95.9 -26.6 -4.3 -12
AAATCCTGCAAAATGTCAAA
703 SEQ ID NO: 3484 -13.3 -28.6 75.5 -15.3 0 -4.9
ATCCTTAATTTTGGGTTTAG
890 SEQ ID NO: 3485 -13.3 -27.8 75.6 -12.3 -2.2 -9.8
GGCAGTGTTACATTACTGGG
934 SEQ ID NO: 3486 -13.3 -36.1 87.3 -18.2 -4.6 -10.9
GATCCTGCTGTTGAGAAAGG
1152 SEQ ID NO:3487 -13.3 -36.2 85.4 -22.3 -0.3 -4.3
TAACCTCCAACAGTGACACC
1728 SEQ ID NO: 3488 -13.3 -37.3 88.3 -24 0 -5.7
ACACAGCAGGTTTGCACTTG
1979 SEQ ID NO: 3489 -13.3 -36.4 87.6 -20.9 -1.7 -12.3
ATCAGGAGCAAAACACAGCA
1991 SEQ ID NO:3490 -13.3 -35.8 85.7 -21.2 -1.2 -3.7
GAGTTTTGTCAGTTGATAAA
2276 SEQ ID NO:3491 -13.3 -28 74.8 -12 -2.5 -12.8
GGTCTTATCCAAAAATGTTT
2345 SEQ ID NO: 3492 -13.3 -27.8 74.8 -13.3 -1.1 -4.6
TTTTTTTTTTTTTTTTTGAC
3215 SEQ ID NO: 3493 -13.3 -17 56.6 -3.7 0 0
TAGTTTACCAGCTTTCTTGC
3433 SEQ ID NO: 3494 -13.3 -32.5 82.9 -18.7 -0.2 -4
TGGAATCACAACTTTTAAGA
3491 SEQ ID NO:3495 -13.3 -29.1 76.5 -15.8 0 -3.5
TTCTTATTTTACACTATACA
4100 SEQ ID NO: 3496 -13.3 -25.2 71.2 -11.9 0 -0.4
ATTCTTATTTTACACTATAC
4101 SEQ ID NO: 3497 -13.3 -24.2 69.5 -10.9 0 -0.4
TTCCTCTGTAATCTCACTGG
4327 SEQ ID NO: 3498 -13.3 -35.4 84.6 -21.2 -0.7 -4.8
AAGGGGAGATTGAGTAAACT
4413 SEQ ID NO:3499 -13.3 -33.9 83.5 -20.6 0 -3.3
GTGCAAGGGGAGATTGAGTA
4417 SEQ ID NO: 3500 -13.3 -37.6 87.9 -24.3 0 -4.8
AAAACAAAAACATGTCCTCA
4686 SEQ ID NO: 3501 -13.3 -28.5 75.7 -15.2 0 -6.2
AAGCACACTGCTGGGGTTTT
170 SEQ ID NO: 3502 -13.2 -37.5 89.6 -22.1 -2.2 -9.3
GAAGTCCATCACATCTCCCC
200 SEQ ID NO: 3503 -13.2 -39.1 88 -25.9 0 -2
TCCCCAGAGAAGTCAAGTTG
1251 SEQ ID NO: 3504 -13.2 -37.2 87.3 -24 0 -3.8
CCATTAGAAAAAACTGTTCG
1288 SEQ ID NO: 3505 -13.2 -27.9 75.4 -14.7 0 -3.7
AGCCATTAGAAAAAACTGTT
1290 SEQ ID NO:3506 -13.2 -28.6 76.7 -15.4 0 -1.4
TTGAATAGCCATTAGAAAAA
1296 SEQ ID NO:3507 -13.2 -26.9 73.4 -13.7 0 -2.8
GCGGCATGCTGGGCAGTTTT
1544 SEQ ID NO: 3508 -13.2 -40.4 92.4 -22.3 -1.8 -17.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
AGGAGTACTGCAGTAGGGTC
1911 SEQ ID NO:3509 -13.2 -40.1 91.8 -23.1 0.1 -15.8
GAGGAAACATACAGCATGTG
2065 SEQ ID NO: 3510 -13.2 -34 82.8 -20 -0.6 -7.2
ACTGAAGAGAGAAGCAGTAA
2137 SEQ ID NO:3511 -13.2 -34.1 83.4 -19.4 -1.4 -8
TATACACTTGTAAACTTTTT
2921 SEQ ID NO: 3512 -13.2 -24.4 70.6 -10.6 -0.3 -6.3
CCTTTTCTGATATACACTTG
2931 SEQ ID NO:3513 -13.2 -29.4 77.1 -16.2 0 -1.6
GCTGTGTTACAGCTGGTTAT
3512 SEQ ID NO: 3514 -13.2 -35 85.8 -16.5 -5.1 -18.2
AGTGAAGAAATTCACAGGAC
3774 SEQ ID NO:3515 -13.2 -32.7 80.8 -12.5 -7 -14
AAATCAAATTTCTTGTGGCT
3890 SEQ ID NO:3516 -13.2 -28.6 75.6 -15.4 0 -3.7
CAGAGGATAACTTCCTCTGT
4338 SEQ ID NO:3517 -13.2 -35.4 84.7 -117 -10.5 -23.4
CCTCTATGCAAACTATTGCC
4571 SEQ ID NO: 3518 -13.2 -34 83.8 -18.9 -1.9 -6.6
ACTGAAAATAACAAATAACA
4724 SEQ ID NO: 3519 -13.2 -24.4 69.5 -11.2 0 -0.6
ATCAGTGAATATCAACTCTG
115 SEQ ID NO:3520 -13.1 -31 77.6 -17.9 0 -6.7
ATGCTTTCTTCCAAAAGCTT
490 SEQ ID NO:3521 -13.1 -30.8 79.4 -14.4 -3.3 -8.8
TCCTCATTCGAGTTTCCTTC
844 SEQ ID NO: 3522 -13.1 -34.6 82.8 -20.1 -1.3 -7.3
CCTATTCCAATTTTCGGAAC
1211 SEQ ID NO:3523 -13.1 -30.7 78.8 -15.1 -2.5 -7.8
CTGCTGAATTCCTTTTATTT
1619 SEQ ID NO: 3524 -13.1 -27.7 74.6 -14.6 0 -5.2
TGGAACAGAGCTATCATATC
1772 SEQ ID NO: 3525 -13.1 -33.8 81.7 -20.7 0 -5.2
CTTGCCGCCCTCCTAACATG
1824 SEQ ID NO: 3526 -13.1 -39.7 91.3 -26.6 0 -3.9
GCTGCAATCACTTGCCGCCC
1834 SEQ ID NO: 3527 -13.1 -41.9 93.3 -23.9 -4.9 -12.6
TGCTGCAATCACTTGCCGCC
1835 SEQ ID NO:3528 -13.1 -40.7 92 -22.7 -4.9 -12.6
TGCACTTGATTGTCTATATG
1967 SEQ ID NO:3529 -13.1 -30.9 78.4 -17.8 0 -4.9
TTGCACTTGATTGTCTATAT
1968 SEQ ID NO: 3530 -13.1 -29.7 76.8 -16.6 0 -4.9
CTGATTGGTGATGATTTCAG
2393 SEQ ID NO: 3531 -13.1 -31.7 78.5 -17.5 -1 -7.2
TGGGATATATTAACTAATAA
2693 SEQ ID NO: 3532 -13.1 -25.8 71.8 -12.7 0 -6.1
CCAACAGAAAGTCTAGAAAA
2843 SEQ ID NO: 3533 -13.1 -30.1 78.5 -16.5 0 -7.5
AAAGGCACAACTTCCCTTTT
2945 SEQ ID NO: 3534 -13.1 -32.8 83.3 -18.5 -1.1 -6.4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
GTAGTTCACTAAATATAAAG
2997 SEQ ID NO: 3535 -13.1 -25.2 71.6 -11.6 -0.2 -5.6
CCAACCCATTTTCACACAGA
3127 SEQ ID NO: 3536 -13.1 -34.5 84.2 -21.4 0 0
GAAAGATTAACCAATTGGTG
3642 SEQ ID NO: 3537 -13.1 -29 76.5 -12.2 -2.2 -15.6
CATCTAAAAATCATTCTTAT
4113 SEQ ID NO: 3538 -13.1 -23.9 67.5 -10.8 0 -0.2
TGCCTGTATGAATGCATGTA
4216 SEQ ID NO: 3539 -13.1 -34.3 83.5 -18.3 -2.7 -13.4
AGCACACTGCTGGGGTTTTC
169 SEQ ID NO: 3540 -13 -39 90.7 -24.1 -1.9 -8.1
AAATCAACCAAAAGTCTTCG
307 SEQ ID NO: 3541 -13 -29 76.3 -16 0 -3.3
AATCCCAGGTCATTTCCCAT
424 SEQ ID NO: 3542 -13 -36.2 85.4 -22.5 -0.4 -3.4
AACTCCTTCTCTGTGGGGGC
580 SEQ ID NO: 3543 -13 -41.8 93.4 -23.7 -5.1 -13
TGTCATAGTGGTACATCTGT
1119 SEQ ID NO: 3544 -13 -34.5 83.7 -20.8 -0.2 -8.4
CTGATCCTGCTGTTGAGAAA
1154 SEQ ID NO:3545 -13 -35 83.8 -22 0 -3.4
AAATAGGCTTCTGATCCTGC
1164 SEQ ID NO: 3546 -13 -35.3 84.6 -20.7 -1.6 -9.8
TATCGATGATGCAATCATTC
1512 SEQ ID NO: 3547 -13 -30.7 76.1 -15.5 -17 -12.4
ATAACCTCCAACAGTGACAC
1729 SEQ ID NO: 3548 -13 -35.1 84.9 -22.1 0 -4.1
AGGTTCAATAACCTCCAACA
1736 SEQ ID NO: 3549 -13 -33.7 83.6 -18.2 -2.5 -8.4
TTTGCACTTGATTGTCTATA
1969 SEQ ID NO: 3550 -13 -29.5 77 -16.5 0 -4.9
GTAACTCAGAGGAAACATAC
2073 SEQ ID NO: 3551 -13 -32.1 80.8 -18.6 0 -7.8
TTATTAAGGCAGTCACTTTT
2459 SEQ ID NO: 3552 -13 -29 77.5 -16 0 -6.2
AATAAAAAATAAAACAACAA
2551 SEQ ID NO:3553 -13 -19.2 60.6 -6.2 0 0
AACAATAAAAAATAAAACAA
2554 SEQ ID NO: 3554 -13 -19.2 60.6 -6.2 0 0
AAACAATAAAAAATAAAACA
2555 SEQ ID NO: 3555 -13 -19.2 60.6 -6.2 0 0
CAGCCAACTGTGAAAAAAAG
2816 SEQ ID NO:3556 -13 -30.4 79.5 -16.8 -0.3 -3.2
ACCAACAGAAAGTCTAGAAA
2844 SEQ ID NO: 3557 -13 -31.4 80.3 -17.9 0 -7.5
CCATGCATAAATCAAAAATG
2892 SEQ ID NO:3558 -13 -26.6 72.4 -13.6 0 -7
CTTAAACAGCTGTACAATAA
3054 SEQ ID NO: 3559 -13 -28 76.4 -12.4 0 -13.3
AGATGATTGATTAATGTTTA
3110 SEQ ID NO: 3560 -13 -25.2 69.2 -11.7 0 -7.5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
GTTAGAAGCACCAACCCATT 3137 SEQ ID NO: 3561 -13 -34.7 85.2 -21.7 0 -2.7
AGTGCCATCAGGTTAGAAGC 3148 SEQ ID NO: 3562 -13 -37.4 87.9 -22.8 -1.5 -5.5
AAATAGTTTACCAGCTTTCT 3436 SEQ ID NO: 3563 -13 -29 77.5 -16 0 -4
AAAAATATATTAGAAGTTCC 3971 SEQ ID NO: 3564 -13 -23.9 68.6 -10.9 0 -5.1
AGTAGCCCTTCCCTTCCCAG 4038 SEQ ID NO: 3565 -13 -41.8 94.3 -28.8 0 -1.3
AAATAACATTCAAAAAGCAT
4712 SEQ ID NO: 3566 -13 -24.1 68.5 -11.1 0 -2.7 CAAATAACATTCAAAAAGCA
4713 SEQ ID NO: 3567 -13 -25.1 70.7 -12.1 0 -2.7 TAGCTTGTGAACGCAGAAGG
35 SEQ ID NO: 3568 -12.9 -36.3 87.1 -20.7 -2.7 -9.7
ATAAACAACTTAGCTTGTGA 45 SEQ ID NO: 3569 -12.9 -29.1 77.4 -15.5 -0.5 -7.1
CTTTTGGAAAATCAACCAAA 315 SEQ ID NO: 3570 -12.9 -27.1 74 -11.8 -2.4 -6.7
CTGATCTCCAAGGACTCTCA 759 SEQ ID NO: 3571 -12.9 -37.9 86.6 -24.5 -0.1 -7.1
GTCCTCATTCGAGTTTCCTT 845 SEQ ID NO: 3572 -12.9 -34.4 82.9 -20.1 -1.3 -7.6
GGCAGTTTTTTCTTCGAATT 1533 SEQ ID NO: 3573 -12.9 -29 76.5 -16.1 0 -5.9
TTCTTTGTTTTTCGAGCTTC 1597 SEQ ID NO: 3574 -12.9 -29.1 76.5 -16.2 0 -6.6
CATATCCTGCATATAACACT 1758 SEQ ID NO: 3575 -12.9 -31.2 79.2 -18.3 0 -4.5
CACAGCAGGTTTGCACTTGA 1978 SEQ ID NO: 3576 -12.9 -36.6 87.4 -21 -2.6 -12.5
CTGAAGAGAGAAGCAGTAAG 2136 SEQ ID NO:3577 -12.9 -34 83.1 -21.1 0 -4.4
TCATCAAATAGCTCTTGGCT 2176 SEQ ID NO: 3578 -12.9 -34.1 83 -19.7 -1.4 -7.1
CACAGACTTTGGGCACTGGT 2776 SEQ ID NO: 3579 -12.9 -39 90.5 -25.3 -0.6 -7.3
GGCACAACTTCCCTTTTCTG 2942 SEQ ID NO: 3580 -12.9 -35.5 86 -22.6 0 -2.4
ATTGTTAAAACCAGACAGTA 2974 SEQ ID NO: 3581 -12.9 -29.1 77.4 -13.8 -2.4 -7.2
ACTTGAATAAAAAATTATGT 3035 SEQ ID NO: 3582 -12.9 -22.1 65.3 -9.2 0 -0.9
CTAGCTGCCCATCTTAAACA 3066 SEQ ID NO: 3583 -12.9 -35 85.7 -22.1 0 -6.3
CAACCCATTTTCACACAGAT 3126 SEQ ID NO: 3584 -12.9 -32.3 80.7 -19.4 0 -1
ACCAACCCATTTTCACACAG 3128 SEQ ID NO: 3585 -12.9 -34.3 84.3 -21.4 0 0
GTGCCATCAGGTTAGAAGCA 3147 SEQ ID NO:3586 -12.9 -37.4 87.9 -22.8 -1.7 -6.6 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
ATGTCTGACATACAGTTCTA
3702 SEQ ID NO:3587 -12.9 -32.3 80.3 -17.7 -0.4 -11.5
ATTTTCAACAGTGAAGAAAT
3783 SEQ ID NO:3588 -12.9 -26.5 71.8 -11 -2.6 -10.4
TGGTAAGGTGCACACAGAAA
3835 SEQ ID NO: 3589 -12.9 -35.8 86.9 -21.4 0 -11
GTCTCCATCTCCCTCTTCCC
4270 SEQ ID NO: 3590 -12.9 -41.5 91 -28.6 0 0
AGTCTCCATCTCCCTCTTCC
4271 SEQ ID NO: 3591 -12.9 -40.3 89.7 -27.4 0 0
TGAGTAAACTAAACCTGCGC
4403 SEQ ID NO: 3592 -12.9 -34.1 85 -21.2 0 -5.1
CAAACTATTGCCAAACTTCT
4563 SEQ ID NO: 3593 -12.9 -29.8 78.4 -16.9 0 -2.9
ACAACTTAGCTTGTGAACGC
41 SEQ ID NO: 3594 -12.8 -33.8 84.2 -20.5 -0.2 -5.9
AACAACTTAGCTTGTGAACG
42 SEQ ID NO: 3595 -12.8 -31.3 80.8 -17.8 -0.5 -7.1
CTCTCCTGAGCAAGCACACT
181 SEQ ID NO:3596 -12.8 -39.5 90 -25.3 -1.3 -5.7
TCTCTTTACCTGGGGACCCA
735 SEQ ID NO: 3597 -12.8 -40.9 93 -24.3 -3.5 -15.1
AAACAGTTTTCATCTATCAA
784 SEQ ID NO: 3598 -12.8 -26.9 72.6 -14.1 0 -2
GAAAGGGATGCTGTATTCAT
1138 SEQ ID NO:3599 -12.8 -33.1 81.2 -17.7 -0.4 -13.4
GGTAATTGTGCTGTCCTTCC
1479 SEQ ID NO: 3600 -12.8 -36.2 86.5 -22.7 -0.5 -6
CCAGGATTTTCAGAGGTTTC
1660 SEQ ID NO: 3601 -12.8 -337 82.5 -20.9 0 -5.8
ACTGCAGTAGGGTCATTTGG
1905 SEQ ID NO:3602 -12.8 -37.1 88.1 -22.1 0 -12.5
CATAAGAAACATCCAGGAGT
1925 SEQ ID NO: 3603 -12.8 -32.9 81.4 -19.6 0 -7.6
TAGTGCGTATTTAAAACAAA
2586 SEQ ID NO: 3604 -12.8 -25.9 73.2 -12.2 0 -9.6
ATATACACTTGTAAACTTTT
2922 SEQ ID NO: 3605 -12.8 -24.6 70.5 -11.2 -0.3 -6.3
AGCAAGCGTAGTTCACTAAA
3004 SEQ ID NO: 3606 -12.8 -32.9 83.4 -19.6 -0.2 -5.6
GAACTAGCTGCCCATCTTAA
3069 SEQ ID NO: 3607 -12.8 -35.3 85.7 -22.5 0 -6.3
GGTTAGAAGCACCAACCCAT
3138 SEQ ID NO: 3608 -12.8 -37.1 88.3 -22.8 -1.4 -6.9
AGGTTAGAAGCACCAACCCA
3139 SEQ ID NO:3609 ' -12.8 -38.1 90.3 -23.7 -1.6 -7.3
CCTGAAAGACAAATAGTTTA
3446 SEQ ID NO: 3610 -12.8 -28 75.6 -14.5 -0.5 -3.1
AAATCATTCTTATTTTACAC
4106 SEQ ID NO: 3611 -12.8 -23.6 67.7 -10.8 0 0
CATGTCATGATAAAACAATC
4134 SEQ ID NO: 3612 -12.8 -27.3 72.5 -12.5 0 -12.2 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CCCTACAAAAAATATATAAC
4153 SEQ ID NO: 3613 -12.8 -25 71.6 -12.2 0 -3.4
CCTCTGTAATCTCACTGGGT
4325 SEQ ID NO: 3614 -12.8 -37.6 87.8 -22.9 -1.9 -7.2
AATAACATTCAAAAAGCATT
4711 SEQ ID NO:3615 -12.8 -24.1 68.5 -11.3 0 -2.7
AACTGCTTTGGACAGATCTG
359 SEQ ID NO: 3616 -127 -34.8 83.9 -20.5 -1 -11.2
TCCAAAAGCTTTAAGTCTGT
481 SEQ ID NO: 3617 -12.7 -31.1 80.4 -17.8 0 -8.6
TCAGAGTGAGTTTTTGGAAA
598 SEQ ID NO:3618 -12.7 -31.2 79.4 -16.6 -1.9 -6.2
CAGCTTCCACAAGTTAAGAC
1435 SEQ ID NO: 3619 -127 -33.6 83.5 -19.7 -1.1 -5.7
CGAATTTTATCGATGATGCA
1519 SEQ ID NO: 3620 -12.7 -29.3 75.1 -15 -0.8 -11
ACACCAGGGTAGGGGTGAGT
1713 SEQ ID NO:3621 -12.7 -43 95.9 -25.3 -5 -13
TCCAACAGTGACACCAGGGT
1723 SEQ ID NO: 3622 -12.7 -40.5 92.1 -27.2 0 -8.5
GGTTTGCACTTGATTGTCTA
1971 SEQ ID NO: 3623 -12.7 -32.6 81.7 -19.9 0 -4.7
TCAGGAGCAAAACACAGCAG
1990 SEQ ID NO: 3624 -127 -36.8 87.3 -22.8 -1.2 -3.6
TTCATAAGATACCTGAAGCC
2096 SEQ ID NO: 3625 -12.7 -33.3 82.2 -20.6 0 -2.8
CTTATCCAAAAATGTTTGGA
2342 SEQ ID NO: 3626 -12.7 -27.7 74.6 -10.9 -4.1 -13.2
ACAGAAGTTTTTTGATATTT
2433 SEQ ID NO: 3627 -12.7 -24.2 687 -10.9 -0.3 -7.9
AACAGAAGTTTTTTGATATT
2434 SEQ ID NO:3628 -12.7 -24.2 68.7 -10.9 -0.3 -7.9
AAACAGAAGTTTTTTGATAT
2435 SEQ ID NO: 3629 -127 -24.2 68.7 -10.9 -0.3 -7.9
TTCCATGCATAAATCAAAAA
2894 SEQ ID NO: 3630 -12.7 -26.7 72.4 -14 0 -7
GCGTAGTTCACTAAATATAA
2999 SEQ ID NO:3631 -12.7 -28 75.9 -14.8 -0.2 -5.6
ACAATAACTTGAATAAAAAA
3041 SEQ ID NO:3632 -12.7 -21.9 65.3 -9.2 0 -1.3
CAGGTTAGAAGCACCAACCC
3140 SEQ ID NO: 3633 -127 -38.1 89.8 -23.8 -1.6 -7.3
TCAGGTTAGAAGCACCAACC
3141 SEQ ID NO:3634 -12.7 -37.2 88.4 -22.9 -1.6 -6
ATCAGGTTAGAAGCACCAAC
3142 SEQ ID NO:3635 -12.7 -35 85 -20.7 -1.6 -6
ACATACAGTTCTAAATCACA
3695 SEQ ID NO: 3636 -127 -29.6 77 -16.9 0 -1.9
GACATACAGTTCTAAATCAC
3696 SEQ ID NO: 3637 -127 -29.9 77.2 -17.2 0 -1.9
CTGACATACAGTTCTAAATC
3698 SEQ ID NO: 3638 -12.7 -29.8 77.1 -16.5 -0.3 -3 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
TAAAATAATTTTCAACAGTG 3790 SEQ ID NO: 3639 -12.7 -23.1 67.6 -10.4 0 -2.1
GATAAAACAATCTCATCTAA 4126 SEQ ID NO: 3640 -12.7 -26.5 71.5 -13.8 0 -1.6
AAAATATATAACATGTCATG 4145 SEQ ID NO: 3641 -12.7 -24.1 68.2 -10.9 0 -8.2
GATGGTGCCTTTAAGGATGT 4482 SEQ ID NO: 3642 -12.7 -35.1 84.8 -21.3 -0.7 -9.8
AAAGTAACCCGCTATTAGAT 4499 SEQ ID NO: 3643 -127 -31.1 80.5 -18.4 0 -4.1
AAACTTCTGAAGCTTCTGTT 4551 SEQ ID NO: 3644 -12.7 -31 79.5 -16.1 0 -12.5
AACTATTGCCAAACTTCTGA 4561 SEQ ID NO: 3645 -12.7 -31.3 80.1 -18.6 0 -2.6
AAGCATTCAAAGAAAACAAA 4698 SEQ ID NO: 3646 -12.7 -26 71.9 -13.3 0 -27
ACATTCAAAAAGCATTCAAA 4707 SEQ ID NO: 3647 -127 -26.2 71.7 -13.5 0 -2.7
ATTAAATAATTTCTCCAAAA 4745 SEQ ID NO: 3648 -12.7 -22.2 65.2 -9.5 0 -0.4
AGCTTGTGAACGCAGAAGGA 34 SEQ ID NO: 3649 -12.6 -37.4 87.9 -22.1 -2.7 -97
ATCAGAGTGAGTTTTTGGAA 599 SEQ ID NO: 3650 -12.6 -31.4 79.1 -16.6 -2.2 -6.5
AGGCTTGCAGTCCTCATTCG 854 SEQ ID NO: 3651 -12.6 -38.7 89 -23.9 -1.1 -12.5
AACCAGATCTCCATTATCCT 905 SEQ ID NO: 3652 -12.6 -34.5 82.8 -21.9 0 -6.5
TCCATAATGACATCCTGAAG 1415 SEQ ID NO: 3653 -12.6 -33.3 81 -207 0 -4.6
TTTCTTCGAATTTTATCGAT 1525 SEQ ID NO: 3654 -12.6 -257 69.7 -10.9 -2.2 -7.1
CAGGTTCATTCCAGCCTGAA 1577 SEQ ID NO: 3655 -12.6 -37.1 87.1 -18.1 -6.4 -15.2
GGTAACGTTGCAGGAACTAT 1690 SEQ ID NO: 3656 -12.6 -34.3 84.8 -18.5 0 -14.6
TATCATATCCTGCATATAAC 1761 SEQ ID NO: 3657 -12.6 -29.6 76.4 -17 0 -4.5
AGAGGAAACATACAGCATGT 2066 SEQ ID NO: 3658 -12.6 -34 83.1 -21.4 0 -5.8
AAAAATGTTTGGAAGCAATA 2335 SEQ ID NO: 3659 -12.6 -26.3 72.7 -12.9 -0.6 -7.7
TGATGATTTCAGCTAACATC 2385 SEQ ID NO: 3660 -12.6 -31 77.7 -17.8 -0.3 -5.9
ATCTGATTGGTGATGATTTC 2395 SEQ ID NO: 3661 -12.6 -31 76.8 -17.9 -0.1 -5.4
TAAATTTCACCATCTACTCT 2676 SEQ ID NO: 3662 -12.6 -29.7 77.1 -17.1 0 -0.9
ATTTCTGGGATATATTAACT 2698 SEQ ID NO: 3663 -12.6 -27.7 74 -15.1 0 -6.8
TTGAATAAAAAATTATGTAA 3033 SEQ ID NO: 3664 -12.6 -20 61.6 -7.4 0 -2.4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CTGGAATCACAACTTTTAAG
3492 SEQ ID NO:3665 -12.6 -28.8 76.6 -16.2 0 -3.6
TGACATACAGTTCTAAATCA
3697 SEQ ID NO: 3666 -12.6 -29.8 76.9 -17.2 0 -1.9
ACAAACATGGATGTCTGACA
3712 SEQ ID NO:3667 -12.6 -33.9 82.1 -20.2 -1 -8.8
GTGAAGAAATTCACAGGACT
3773 SEQ ID NO:3668 -12.6 -32.7 80.8 -13.3 -6.8 -16.5
AAATATATAACATGTCATGA
4144 SEQ ID NO: 3669 -12.6 -25.6 69.9 -12 0 -9.9
GAATTAGTGTATTATTGGCA
4628 SEQ ID NO: 3670 -12.6 -28.6 76.1 -16 0 -4.3
AACGCAGAAGGAGCAGGAGG
26 SEQ ID NO:3671 -12.5 -40.9 92.2 -27.3 -1 -3.3
AGGAGTTAATGATTCTTTGG
140 SEQ ID NO: 3672 -12.5 -30.4 78.1 -157 -1.8 -12.2
CCCTCTCCTGAGCAAGCACA
183 SEQ ID NO: 3673 -12.5 -41.8 93 -27.7 -1.6 -5.9
CTTCTCTGTGGGGGCAGCAG
575 SEQ ID NO:3674 -12.5 -42.7 94.3 -27.9 -2.2 -11.9
CCTTCTCTGTGGGGGCAGCA
576 SEQ ID NO:3675 -12.5 -43.9 96 -27.2 -4.2 -12
ACATCAGAGTGAGTTTTTGG
601 SEQ ID NO:3676 -12.5 -32.4 80.8 -18.3 -1.5 -5.3
GGCTTGACAAAACCAGATCT
915 SEQ ID NO:3677 -12.5 -34.8 84.2 -21.5 -0.6 -7.4
ATAGCCATTAGAAAAAACTG
1292 SEQ ID NO:3678 -12.5 -27.9 75.4 -15.4 0 -1.3
TGTGCTGTCCTTCCACTGCT
1473 SEQ ID NO:3679 -12.5 -40.5 91.9 -26.8 -1.1 -4.6
AATTTTATCGATGATGCAAT
1517 SEQ ID NO:3680 -12.5 -26.5 70.8 -12.5 0 -11
TGATTGTCTATATGATCTCC
1961 SEQ ID NO:3681 -12.5 -31.8 78.2 -177 -1.6 -8.2
AAACACAGCAGGTTTGCACT
1981 SEQ ID NO: 3682 -12.5 -35.2 86.3 -20.4 -1.7 -12.5
TCATAAGATACCTGAAGCCT
2095 SEQ ID NO:3683 -12.5 -34.5 83.8 -22 0 -1.5
CATTCTAATTTCATCAAATA
2186 SEQ ID NO: 3684 -12.5 -23.9 67.4 -11.4 0 -0.9
TACTCATGGTCTTATCCAAA
2352 SEQ ID NO:3685 -12.5 -32.1 80.5 -17.5 -2.1 -6.8
GCCAACTGTGAAAAAAAGTA
2814 SEQ ID NO:3686 -12.5 -29.7 787 -17.2 0 -3.7
AGCCAACTGTGAAAAAAAGT
2815 SEQ ID NO: 3687 -12.5 -30.5 79.7 -18 0 -1.5
AATTGTTAAAACCAGACAGT
2975 SEQ ID NO:3688 -12.5 -28.7 76.7 -14.2 -2 -6.4
ATTAATGTTTAGAGTTTATT
3101 SEQ ID NO:3689 -12.5 -22.5 66.2 -10 0 -3.8
AGATGGGAATGTGAAAATGG
3620 SEQ ID NO: 3690 -12.5 -32.6 80.4 -20.1 0 0 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
AGTTCTAAATCACAAAAATC
3689 SEQ ID NO: 3691 -12.5 -25.7 70.9 -13.2 0 -1
TATAAAAATATATTAGAAGT
3974 SEQ ID NO:3692 -12.5 -21 63.5 -8.5 0 -5.1
TACCAATATGATATATATCT
4009 SEQ ID NO: 3693 -12.5 -26.5 71.5 -12.4 0 -11.4
AATATATAACATGTCATGAT
4143 SEQ ID NO: 3694 -12.5 -25.8 69.9 -12.3 0 -9.9
AAAGGAATTAGTGTATTATT
4632 SEQ ID NO: 3695 -12.5 -24.9 70.4 -12.4 0 -4.3
AATGCTTTCTTCCAAAAGCT
491 SEQ ID NO: 3696 -12.4 -30.8 79.4 -15.1 -3.3 -8.8
GACTCTCATTCGTCTCTTTA
747 SEQ ID NO: 3697 -12.4 -327 80.2 -18.7 -1.6 -4.5
AGCACATAGGTAATTGTGCT
1487 SEQ ID NO:3698 -12.4 -33.9 84 -14.1 -7.4 -167
CTTCGAATTTTATCGATGAT
1522 SEQ ID NO:3699 -12.4 -27.1 71.6 -12.3 -2.4 -11
AACTATTGTTTTGTTACCAG
1676 SEQ ID NO:3700 -12.4 -27.6 75.7 -15.2 0 -0.8
GTAACGTTGCAGGAACTATT
1689 SEQ ID NO:3701 -12.4 -31.9 81.5 -16.1 0 -14.9
CAGGAGTACTGCAGTAGGGT
1912 SEQ ID NO: 3702 -12.4 -39.8 91.9 -23.5 -1.4 -16
TATCCAAAAATGTTTGGAAG
2340 SEQ ID NO: 3703 -12.4 -27.7 74.6 -10.4 -4.9 -15
ACTCATGGTCTTATCCAAAA
2351 SEQ ID NO: 3704 -12.4 -31.7 79.8 -17.5 -1.8 -6.5
TGAAAACAATAAAAAATAAA
2558 SEQ ID NO:3705 -12.4 -19.4 60.6 -7 0 0
TTTTCTGATATACACTTGTA
2929 SEQ ID NO:3706 -12.4 -27.5 74.2 -15.1 0 -4.5
AAGTGCCATCAGGTTAGAAG
3149 SEQ ID NO:3707 -12.4 -34.9 84.9 -20.9 -1.5 -5.5
CAGTTCTAAATCACAAAAAT
3690 SEQ ID NO: 3708 -12.4 -25.4 70.8 -13 0 -1.9
TACAGTTCTAAATCACAAAA
3692 SEQ ID NO: 3709 -12.4 -26.9 73.6 -14.5 0 -1.9
TACAAACATGGATGTCTGAC
3713 SEQ ID NO:3710 -12.4 -33.1 81.3 -19.4 -1 -9.9
TCAACAGTGAAGAAATTCAC
3779 SEQ ID NO:3711 -12.4 -30.3 77.4 -12.6 -5.3 -10.6
GAATAGAAATCAAATTTCTT
3896 SEQ ID NO:3712 -12.4 -23.6 66.9 -10.1 -1 -7.6
TCCATATTTTTAATATTAGA
3954 SEQ ID NO:3713 -12.4 -23.2 66.9 -10.8 0 -7
TCATCTAAAAATCATTCTTA
4114 SEQ ID NO: 3714 -12.4 -25.2 69.3 -12.8 0 0
TGTCATGATAAAACAATCTC
4132 SEQ ID NO:3715 -12.4 -28.6 74.3 -14.6 0 -11.4
ATGCATGTACAAACTATAAA
4205 SEQ ID NO: 3716 -12.4 -27.2 74.1 -14.1 0 -9.1 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecu Site oligo binding formation Duplex structure oligo oligo
AATGCATGTACAAACTATAA
4206 SEQ ID NO:3717 -12.4 -27.2 74.1 -14.1 0 -9.1
GAGTAAACTAAACCTGCGCT
4402 SEQ ID NO:3718 -12.4 -34.1 85.1 -21.2 0 -7.5
CAAGGGGAGATTGAGTAAAC
4414 SEQ ID NO:3719 -12.4 -33.9 83.3 -21.5 0 -2
TGTATTATTGGCAACCTATG
4621 SEQ ID NO: 3720 -12.4 -307 79.5 -18.3 0 -4.9
ATTCAAAAAGCATTCAAAGA
4705 SEQ ID NO:3721 -12.4 -26.4 71.7 -14 0 -27
ACAAATAACATTCAAAAAGC
4714 SEQ ID NO: 3722 -12.4 -25.2 70.9 -12.8 0 0
TTTGGAAAATCAACCAAAAG
313 SEQ ID NO: 3723 -12.3 -27.1 74 -11.8 -3 -7.9
TTCTTCGAATTTTATCGATG
1524 SEQ ID NO: 3724 -12.3 -26.9 71.7 -12.2 -2.4 -7.3
ACTATTGTTTTGTTACCAGG
1675 SEQ ID NO:3725 -12.3 -30 79.3 -17.7 0 -2.4
CTGGAACAGAGCTATCATAT
1773 SEQ ID NO:3726 -12.3 -33.5 81.7 -21.2 0 -6.7
AGCAAATGCCATAAGAAACA
1934 SEQ ID NO: 3727 -12.3 -31.2 79.9 -18.1 -0.6 -6.1
GAGCAAATGCCATAAGAAAC
1935 SEQ ID NO: 3728 -12.3 -31.5 79.9 -18.3 -0.8 -6.3
CATGGTCTTATCCAAAAATG
2348 SEQ ID NO: 3729 -12.3 -29.1 76.2 -14.7 -2.1 -6.8
AAGGCAACCATTCTTATTAA
2472 SEQ ID NO:3730 -12.3 -29.1 77.3 -16.8 0 -3.9
ATAAAAAATAAAACAACAAA
2550 SEQ ID NO: 3731 -12.3 -19.2 60.6 -6.9 0 0
GGGATACACCAACAGAAAGT
2851 SEQ ID NO:3732 -12.3 -35 85.1 -20.7 -2 -7.4
CCTAACTATACAGGGGGGGG
2868 SEQ ID NO:3733 -12.3 -41 94.2 -24.1 -4.6 -13.2
AGCTGGTTATCTGGAATCAC
3502 SEQ ID NO: 3734 -12.3 -35.4 84.5 -21.6 -1.4 -8.1
AATCAAATTTCTTGTGGCTT
3889 SEQ ID NO: 3735 -12.3 -28.6 75.6 -16.3 0 -3.8
AAGTTCCATATTTTTAATAT
3958 SEQ ID NO: 3736 -12.3 -22.6 66.1 -10.3 0 -2.3
ATCTAAAAATCATTCTTATT
4112 SEQ ID NO: 3737 -12.3 -22.7 65.3 -10.4 0 -0.2
CGCTGCCTGTATGAATGCAT
4219 SEQ ID NO:3738 -12.3 -36.6 86.2 -23.3 -0.6 -9.6
AGGACCAGTCTCCATCTCCC
4277 SEQ ID NO:3739 -12.3 -42.5 92.6 -27.8 -2.4 -8.8
TTCAAAAAGCATTCAAAGAA
4704 SEQ ID NO: 3740 -12.3 -26.2 71.8 -13.9 0 -2.7
TATTAAATAATTTCTCCAAA
4746 SEQ ID NO: 3741 -12.3 -22.6 66.1 -10.3 0 -1.4
TGAACGCAGAAGGAGCAGGA
28 SEQ ID NO: 3742 -12.2 -40 90.7 -26.9 -0.7 -3 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TTTTGGAAACTCCTTCTCTG
587 SEQ ID NO: 3743 -12.2 -32.3 81.1 -19 -1 -6.3
ACTGGGGCTTGACAAAACCA
920 SEQ ID NO: 3744 -12.2 -37.7 89.5 -24 -0.8 -10.8
AGAAAGGGATGCTGTATTCA
1139 SEQ ID NO: 3745 -12.2 -34.1 83.1 -19.3 -0.4 -13.4
GCTGAATTCCTTTTATTTTT
1617 SEQ ID NO: 3746 -12.2 -25.3 71 -13.1 0 -5.2
CTCCTAACATGTTGAGCGTA
1815 SEQ ID NO: 3747 -12.2 -34.5 84.4 -20.8 0 -11
TACTGCAGTAGGGTCATTTG
1906 SEQ ID NO:3748 -12.2 -35.1 85.7 -19.3 0 -15.3
ATAAGAAACATCCAGGAGTA
1924 SEQ ID NO:3749 -12.2 -32.1 80.6 -19.2 0 -8.7
AGGTTTGCACTTGATTGTCT
1972 SEQ ID NO: 3750 -12.2 -33.4 82.6 -21.2 0 -4.9
AACACAGCAGGTTTGCACTT
1980 SEQ ID NO: 3751 -12.2 -35.2 86.3 -20.7 -1.7 -12.5
GCAACCATTCTTATTAAGGC
2469 SEQ ID NO:3752 -12.2 -31.6 80.8 -19.4 0 -7.2
CAAAAATCAGTAGCTGAGCT
3677 SEQ ID NO: 3753 -12.2 -32.5 81.6 -17.5 -1.5 -13.8
ACATGGATGTCTGACATACA
3708 SEQ ID NO: 3754 -12.2 -34.5 82.5 -19.9 -0.3 -13
AATATGATATATATCTGAAA
4005 SEQ ID NO: 3755 -12.2 -23.9 66.8 -10.1 0 -11.4
CAATCATTTTAATAAATTGC
4070 SEQ ID NO:3756 -12.2 -22.2 65.7 -10 0 -3.2
ACCTGCGCTGACAGACTCAC
4391 SEQ ID NO: 3757 -12.2 -41.2 91.8 -27.6 -1.3 -7.5
AAGCACCACCTTCCTGTCTC
4462 SEQ ID NO:3758 -12.2 -39.5 90.4 -27.3 0 -2.7
AAGTAACCCGCTATTAGATG
4498 SEQ ID NO:3759 -12.2 -32.3 82 -20.1 0 -4.1
ACTATTGCCAAACTTCTGAA
4560 SEQ ID NO:3760 -12.2 -31.3 80.1 -19.1 0 -2.9
GCACTATTTACATATTGCTG
4594 SEQ ID NO:3761 -12.2 -29.6 77.8 -15.5 -1.9 -8.6
AGCAAGCACACTGCTGGGGT
173 SEQ ID NO: 3762 -12.1 -42.4 95.1 -27.9 -2.4 -9.3
CGACAGCCAGTGAGGGTGAA
264 SEQ ID NO: 3763 -12.1 -41 92 -25 -3.9 -8.9
AAACTGCTTTGGACAGATCT
360 SEQ ID NO:3764 -12.1 -33.6 82.6 -20.3 -1 -9.3
CTCATTCGAGTTTCCTTCCA
842 SEQ ID NO:3765 -12.1 -34.3 82.8 -21.1 -1 -7.6
TTCTGATCCTGCTGTTGAGA
1156 SEQ ID NO: 3766 -12.1 -36.5 85.3 -24.4 0 -3.4
GAATTGGTGGAATGACATTA
1185 SEQ ID NO: 3767 -12.1 -30.7 77.9 -16.9 -1.7 -8.2
ACCTATTCCAATTTTCGGAA
1212 SEQ ID NO: 3768 -12.1 -30.7 78.8 -16.9 -1.7 -6.2 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GCAGTTTTTTCTTCGAATTT
1532 SEQ ID NO:3769 -12.1 -26.6 72.8 -14.5 0 -5.9
ACTAGCTGCCCATCTTAAAC
3067 SEQ ID NO:3770 -12.1 -35.1 85.9 -23 0 -6.3
CAGCTGGTTATCTGGAATCA
3503 SEQ ID NO:3771 -12.1 -35.3 84.4 -21.6 -1.4 -10.3
GGGAATGTGAAAATGGGTGT
3616 SEQ ID NO: 3772 -12.1 -34.7 84.1 -22.6 0 0
AAAATCAGTAGCTGAGCTTT
3675 SEQ ID NO: 3773 -12.1 -31.3 80.1 -16.4 -1.5 -13.8
ACAGTTCTAAATCACAAAAA
3691 SEQ ID NO: 3774 -12.1 -26.5 72.8 -14.4 0 -1.9
ATACAGTTCTAAATCACAAA
3693 SEQ ID NO: 3775 -12.1 -27.1 73.4 -15 0 -1.9
ATAAAAATATATTAGAAGTT
3973 SEQ ID NO: 3776 -12.1 -20.6 62.7 -8.5 0 -4.8
TCTAAAAATCATTCTTATTT
4111 SEQ ID NO: 3777 -12.1 -22.5 65.3 -10.4 0 -0.2
CTGAAAATAACAAATAACAT
4723 SEQ ID NO: 3778 -12.1 -23.3 67.5 -11.2 0 -0.6
GAACGCAGAAGGAGCAGGAG
27 SEQ ID NO:3779 -12 -40 90.4 -26.9 -1 -3.3
TAAACAACTTAGCTTGTGAA
44 SEQ ID NO:3780 -12 -28.9 77.6 -16.2 -0.5 -7.1
ATCCCAGGTCATTTCCCATC
423 SEQ ID NO: 3781 -12 -37.7 86.7 -25 -0.4 -3.4
GCAATGCTTTCTTCCAAAAG
493 SEQ ID NO: 3782 -12 -30.8 79.3 -18.3 0 -7.6
CCAGAGGTACTCACACCATG
1096 SEQ ID NO:3783 -12 -38.6 89 -24.5 -2.1 -8.5
ATGTCATAGTGGTACATCTG
1120 SEQ ID NO: 3784 -12 -33.4 81.7 -21.4 0 -6.5
GTATTCATGTCATAGTGGTA
1126 SEQ ID NO: 3785 -12 -31.4 79.4 -18.5 -0.7 -5.3
AACCTCCAACAGTGACACCA
1727 SEQ ID NO: 3786 -12 -38.1 89.1 -26.1 0 -5.7
TTCTGATATACACTTGTAAA
2927 SEQ ID NO: 3787 -12 -27.5 74.2 -14.9 -0.3 -6.3
CACCAACCCATTTTCACACA
3129 SEQ ID NO:3788 -12 -34.3 84.3 -22.3 0 0
TGCCATCAGGTTAGAAGCAC
3146 SEQ ID NO:3789 -12 -37.4 87.9 -23.8 -1.5 -6.4
CTAAACCTGCGCTGACAGAC
4395 SEQ ID NO: 3790 -12 -37.6 88.4 -24.2 -1.3 -7.5
GGGGAGATTGAGTAAACTAA
4411 SEQ ID NO: 3791 -12 -33.1 82.6 -21.1 0 -5.2
AGTGCAAGGGGAGATTGAGT
4418 SEQ ID NO: 3792 -12 -38.4 88.8 -26.4 0 -4.8
CTCTATGCAAACTATTGCCA
4570 SEQ ID NO:3793 -12 -32.8 82.2 -18.9 -1.9 -6.6
CCTCTCCTGAGCAAGCACAC
182 SEQ ID NO: 3794 -11.9 -40.7 91.3 -27.2 -1.6 -5.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GACAGCCAGTGAGGGTGAAG
263 SEQ ID NO:3795 -11.9 -40.7 91.4 -24.9 -3.9 -8.9
TCCAAATCCTGCAAAATGTC
706 SEQ ID NO: 3796 -11.9 -32.5 80.7 -20.6 0 -4.9
GGCTTGCAGTCCTCATTCGA
853 SEQ ID NO: 3797 -11.9 -39 88.9 -25.4 -0.9 -11.5
ACATTAAAAATAGGCTTCTG
1171 SEQ ID NO: 3798 -11.9 -27.9 75.4 -16 0 -5
GGATGGAGGAGAGCTTACAT
1334 SEQ ID NO:3799 -11.9 -38 87.5 -25.4 -0.4 -7.6
AATAACCTCCAACAGTGACA
1730 SEQ ID NO: 3800 -11.9 -33.8 83.4 -21.9 0 -4.1
CCAAGTTGAGTCTGGAACAG
1784 SEQ ID NO: 3801 -11.9 -35.8 85.7 -22.6 -0.9 -10.1
GAAACAGAAGTTTTTTGATA
2436 SEQ ID NO: 3802 -11.9 -25.5 71 -127 -0.8 -7.9
TGATGAAACAGAAGTTTTTT
2440 SEQ ID NO:3803 -11.9 -26.3 71.9 -13.3 -1 -4.5
CTGATAGTTTATACAATAAA
2514 SEQ ID NO:3804 -11.9 -24.1 69.2 -12.2 0 -3
GGGATATATTAACTAATAAA
2692 SEQ ID NO: 3805 -11.9 -24.6 70.2 -127 0 -6.1
TTTCTGGGATATATTAACTA
2697 SEQ ID NO: 3806 -11.9 -27.9 75 -16 0 -6.8
GAAAGTCTAGAAAATTTCAT
2837 SEQ ID NO: 3807 -11.9 -25.9 70.9 -13 -0.5 -9.8
TTTCTGATATACACTTGTAA
2928 SEQ ID NO:3808 -11.9 -27.5 74.2 -15 -0.3 -6.3
AAAAAATTATGTAAGAAAAA
3027 SEQ ID NO: 3809 -11.9 -19.4 60.7 -7.5 0 -4.2
TGAGCTTTTTTTTTTTTTTT
3221 SEQ ID NO:3810 -11.9 -20.6 63.4 -8.7 0 -6.4
ACTTTTAAGAAGTTATACAA
3481 SEQ ID NO:3811 -11.9 -24.6 70.6 -12.2 0 -7.6
AAAAATCAGTAGCTGAGCTT
3676 SEQ ID NO: 3812 -11.9 -31.3 80.1 -16.7 -1.2 -13.5
TTACAAACATGGATGTCTGA
3714 SEQ ID NO: 3813 -11.9 -31.8 79.6 -18.6 -1 -9.9
AGAAATCAAATTTCTTGTGG
3892 SEQ ID NO: 3814 -11.9 -27.6 73.7 -14.8 -0.8 -6.5
CTTTAAGGATGTAAGCACCA
4474 SEQ ID NO:3815 -11.9 -32.6 82.6 -18.4 -2.3 -9.1
TAACAAATAACATTCAAAAA
4716 SEQ ID NO: 3816 -11.9 -21.9 65.2 -10 0 0
GTGAACGCAGAAGGAGCAGG
29 SEQ ID NO: 3817 -11.8 -39.8 90.5 -26.9 -1 -5.6
TTGTGAACGCAGAAGGAGCA
31 SEQ ID NO: 3818 -11.8 -37.4 87.9 -24.4 -1.1 -8.2
CACATCTCCCCTCTCCTGAG
191 SEQ ID NO:3819 -11.8 -41.2 90.8 -28.1 -1.2 -4.6
TGGAAACTCCTTCTCTGTGG
584 SEQ ID NO: 3820 -11.8 -37.2 87.3 -24.3 -1 -6.3 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TTGGAAACTCCTTCTCTGTG
585 SEQ ID NO: 3821 -11.8 -34.8 84.2 -21.9 -1 -6.3
AGAGGCTTGCAGTCCTCATT
856 SEQ ID NO: 3822 -11.8 -38.4 88.7 -19.9 -5.4 -21.5
ACCAGATCTCCATTATCCTT
904 SEQ ID NO:3823 -11.8 -34.5 82.8 -227 0 -6.5
TTCGAATTTTATCGATGATG
1521 SEQ ID NO: 3824 -11.8 -27.1 71.5 -12.9 -2.4 -11
TTTTCTTCGAATTTTATCGA
1526 SEQ ID NO: 3825 -11.8 -25.5 69.8 -11.9 -1.8 -5.8
ATCATATCCTGCATATAACA
1760 SEQ ID NO:3826 -11.8 -30.4 77.2 -18.6 0 -4.5
ACAGCAGGTTTGCACTTGAT
1977 SEQ ID NO:3827 -11.8 -35.6 85.8 -21 -2.8 -12.5
AAAACAATAAAAAATAAAAC
2556 SEQ ID NO: 3828 -11.8 -18 58.8 -6.2 0 0
AGTGCGTATTTAAAACAAAA
2585 SEQ ID NO: 3829 -11.8 -25.5 72.3 -13.1 0 -8.5
GTTCATCACACAGACTTTGG
2784 SEQ ID NO: 3830 -11.8 -33.7 82.3 -21.9 0 -5.6
TTTCCATGCATAAATCAAAA
2895 SEQ ID NO: 3831 -11.8 -267 72.4 -14.9 0 -6.7
GGTTTCCATGCATAAATCAA
2897 SEQ ID NO:3832 -11.8 -30.4 78 -17 -0.8 -10.9
TGATATACACTTGTAAACTT
2924 SEQ ID NO: 3833 -11.8 -27.3 74.3 -15.5 0 -6
GCTATAAAAGGCACAACTTC
2951 SEQ ID NO:3834 -11.8 -31.5 81 -17.4 -2.3 -6.2
GATGATTGATTAATGTTTAG
3109 SEQ ID NO:3835 -11.8 -25.2 69.7 -13.4 0 -7.4
AAATCACAAAAATCAGTAGC
3683 SEQ ID NO: 3836 -11.8 -27.9 747 -16.1 0 -0.8
GGATGTGTAGTTTTACAAAC
3726 SEQ ID NO: 3837 -11.8 -29.2 77.6 -15.8 -1.6 -8.8
AAGGTGCACACAGAAAGGGC
3831 SEQ ID NO: 3838 -11.8 -39 907 -257 0 -11
AAATCATATAGGTTATCCAT
4188 SEQ ID NO:3839 -11.8 -27.9 73.8 -14.9 -1.1 -5.3
TGCATGTACAAACTATAAAT
4204 SEQ ID NO:3840 -11.8 -27.2 74.1 -14.9 0 -8.2
TAAACCTGCGCTGACAGACT
4394 SEQ ID NO: 3841 -11.8 -37.6 88.8 -24.7 -1 -7.5
ATTAGATGGTGCCTTTAAGG
4486 SEQ ID NO: 3842 -11.8 -32.7 82.4 -18.2 0 -13.6
AGCATTCAAAGAAAACAAAA
4697 SEQ ID NO: 3843 -11.8 -26 71.9 -14.2 0 -2.7
CAAAATACTGAAAATAACAA
4730 SEQ ID NO: 3844 -11.8 -23.1 67.6 -11.3 0 -0.6
AAAGCAAATGATTGTTTTTT
4766 SEQ ID NO: 3845 -11.8 -23.5 67.8 -9.5 -2.2 -7.2
AGCAGGAGGGAAATATATTT
15 SEQ ID NO: 3846 -117 -31.9 80.6 -20.2 0 -5.2 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GGAAACTCCTTCTCTGTGGG
583 SEQ ID NO: 3847 -11.7 -38.4 88.8 -25.5 -1.1 -6.3
AAACCAGATCTCCATTATCC
906 SEQ ID NO:3848 -11.7 -33.3 81.2 -21.6 0 -6.2
TATTCATGTCATAGTGGTAC
1125 SEQ ID NO: 3849 -117 -31.4 79.4 -18.8 -0.7 -6.5
TTCAGAGTCCCCAGAGAAGT
1258 SEQ ID NO: 3850 -11.7 -38.7 88.8 -27 0 -3.1
AACTGTTCGACCAGGGAAGT
1277 SEQ ID NO:3851 -11.7 -37.4 88.2 -23.3 -1.6 -12.7
TGAATAGCCATTAGAAAAAA
1295 SEQ ID NO: 3852 -117 -26.9 73.4 -15.2 0 -2.8
GAACTATTGTTTTGTTACCA
1677 SEQ ID NO: 3853 -11.7 -27.9 75.7 -15.5 -0.4 -3
CAATAACCTCCAACAGTGAC
1731 SEQ ID NO: 3854 -11.7 -33.8 83.2 -22.1 0 -3.2
GTACTGCAGTAGGGTCATTT
1907 SEQ ID NO: 3855 -11.7 -35.2 85.8 -19.6 -0.1 -16
CAAATGCCATAAGAAACATC
1932 SEQ ID NO: 3856 -11.7 -29.2 76.2 -17.5 0 -3.4
TAGCTCTTGGCTCTTCAGAC
2168 SEQ ID NO: 3857 -11.7 -37.7 87.9 -24.4 -1.5 -8
ATTTCATCAAATAGCTCTTG
2179 SEQ ID NO: 3858 -11.7 -28.2 74.3 -16.5 0 -3.8
AGTTCATCACACAGACTTTG
2785 SEQ ID NO: 3859 -11.7 -32.5 80.7 -20.8 0 -3.2
ACACCAACAGAAAGTCTAGA
2846 SEQ ID NO: 3860 -11.7 -33.9 83.5 -22.2 0 -5.6
AAATCAGTAGCTGAGCTTTC
3674 SEQ ID NO:3861 -11.7 -32.8 81.6 -18.3 -1.5 -13.8
CAGTGAAGAAATTCACAGGA
3775 SEQ ID NO:3862 -11.7 -32.6 80.7 -13.9 -7 -14
TTTAATAAATTGCATGTAAA
4063 SEQ ID NO: 3863 -11.7 -22.2 66 -8.9 0 -11.4
TTGACCCCTACAAAAAATAT
4158 SEQ ID NO: 3864 -11.7 -29.1 77.4 -17.4 0 -0.8
AAACCTGCGCTGACAGACTC
4393 SEQ ID NO: 3865 -11.7 -38.7 89.2 -25.6 -1.3 -7.5
AAACTATTGCCAAACTTCTG
4562 SEQ ID NO: 3866 -117 -29.8 78.4 -18.1 0 -2.9
ATCCAAAAATGTTTGGAAGC
2339 SEQ ID NO: 3867 -11.6 -29.8 77.5 -13.3 -4.9 -15
TAAAAAATAAAACAACAAAA
2549 SEQ ID NO:3868 -11.6 -19 60.5 -7.4 0 0
AAATTGTTAAAACCAGACAG
2976 SEQ ID NO:3869 -11.6 -27.4 74.8 -14.9 -0.7 -3.7
GATTAATGTTTAGAGTTTAT
3102 SEQ ID NO:3870 -11.6 -24 68.6 -12.4 0 -3.8
TTATCTGGAATCACAACTTT
3496 SEQ ID NO: 3871 -11.6 -29.3 76.3 -177 0 -6.3
CTCATCTAAAAATCATTCTT
4115 SEQ ID NO: 3872 -11.6 -26 70.6 -14.4 0 0 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecu Site oligo binding formation Duplex structure oligo oligo
AACCTGCGCTGACAGACTCA
4392 SEQ ID NO: 3873 -11.6 -39.9 90.7 -26.9 -1.3 -7.5
TATTATTGGCAACCTATGAG
4619 SEQ ID NO: 3874 -11.6 -30.9 79.5 -18.6 -0.5 -6.5
GAAAACAAAAACATGTCCTC
4687 SEQ ID NO: 3875 -11.6 -28.8 76 -17.2 0 -6.2
TACTGAAAATAACAAATAAC
4725 SEQ ID NO: 3876 -11.6 -23.6 68.7 -12 0 -0.6
TTCCAAAAGCTTTAAGTCTG
482 SEQ ID NO: 3877 -11.5 -29.8 78.6 -17.8 0 -8.3
CATCAGAGTGAGTTTTTGGA
600 SEQ ID NO: 3878 -11.5 -32.6 80.7 -18.9 -2.2 -6.5
GGGCAGTTTTTTCTTCGAAT
1534 SEQ ID NO: 3879 -11.5 -31.4 80 -19.9 0 -6.1
GCTTCCAGGTTCATTCCAGC
1582 SEQ ID NO: 3880 -11.5 -38.4 88.5 -25.5 -1.3 -4.6
GCAAAACACAGCAGGTTTGC
1984 SEQ ID NO: 3881 -11.5 -35.2 86.1 -22.4 -1.2 -9.5
CAGAGGAAACATACAGCATG
2067 SEQ ID NO: 3882 -11.5 -33.9 82.8 -22.4 0 -2.7
ATACCTGAAGCCTGTGTAAC
2088 SEQ ID NO:3883 -11.5 -35.2 85.8 -23.7 0 -2.9
GAGCTTTTTTTTTTTTTTTT
3220 SEQ ID NO:3884 -11.5 -19.4 61.6 -7.9 0 -5.3
GGAAAGATTAACCAATTGGT
3643 SEQ ID NO:3885 -11.5 -30.2 78.4 -16 -1.5 -13.6
TTAAAATAATTTTCAACAGT
3791 SEQ ID NO:3886 -11.5 -21.9 65.3 -10.4 0 -2.1
AAAAATATATAACATGTCAT
4146 SEQ ID NO: 3887 -11.5 -22.9 66 -11.4 0 -6.2
AGGGGAGATTGAGTAAACTA
4412 SEQ ID NO: 3888 -11.5 -34.3 84.2 -22.8 0 -5.2
TTAAGGATGTAAGCACCACC
4472 SEQ ID NO: 3889 -11.5 -35.1 85.9 -21.3 -2.3 -9.1
TTTAAGGATGTAAGCACCAC
4473 SEQ ID NO:3890 -11.5 -32.7 82.7 -19.1 -2.1 -8.9
GATTCTGCACTATTTACATA
4600 SEQ ID NO: 3891 -11.5 -28.9 76 -17.4 0 -4.6
AAAAACATGTCCTCATTTTA
4681 SEQ ID NO: 3892 -11.5 -26.6 72.6 -15.1 0 -6.2
AAGCAAATGATTGTTTTTTT
4765 SEQ ID NO: 3893 -11.5 -23.5 67.8 -10.6 -1.3 -6.1
CACACTGCTGGGGTTTTCTT
167 SEQ ID NO: 3894 -11.4 -36.5 87.7 -25.1 0 -4.6
CTTGCAGTCCTCATTCGAGT
851 SEQ ID NO: 3895 -11.4 -36.6 85.8 -23.8 -1.3 -8.4
TGGTGGAATGACATTAAAAA
1181 SEQ ID NO: 3896 -11.4 -29 76.3 -15.3 -2.3 -9.4
CTTCCAGGTTCATTCCAGCC
1581 SEQ ID NO:3897 -11.4 -38.3 88.6 -25.5 -1.3 -5
TCAATAACCTCCAACAGTGA
1732 SEQ ID NO:3898 -11.4 -34 83.3 -22.6 0 -3.2 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
AATGGCTTTTCCTAGCTCTT
2216 SEQ ID NO: 3899 -11.4 -33.6 83.6 -19.4 -2.8 -87
CTCTCCCATCACTGAAAAGT
2660 SEQ ID NO: 3900 -11.4 -35 83.9 -22.1 -1.4 -4.6
ATAAATCAAAAATGCTATCC
2886 SEQ ID NO:3901 -11.4 -26.1 71.5 -14.7 0 -17
TACGAACTAGCTGCCCATCT
3072 SEQ ID NO: 3902 -11.4 -38.1 89.3 -26.7 0 -6.3
ACACAGATGATTGATTAATG
3114 SEQ ID NO: 3903 -11.4 -28.5 74.1 -17.1 0 -7.4
ACCATCAGGAAAGATTAACC
3650 SEQ ID NO: 3904 -11.4 -32.9 81.6 -20.6 -0.8 -2.9
AATTTTCAACAGTGAAGAAA
3784 SEQ ID NO: 3905 -11.4 -26.3 71.9 -12.3 -2.6 -10.4
ATATAAAAATATATTAGAAG
3975 SEQ ID NO: 3906 -11.4 -19.9 61.6 -8.5 0 -5.1
GTATGAATGCATGTACAAAC
4211 SEQ ID NO: 3907 -11.4 -29.6 77.1 -16.3 -17 -11.3
GCACACAGGACCAGTCTCCA
4283 SEQ ID NO:3908 -11.4 -42 92.9 -28.2 -2.4 -8.8
TATGAGATTCTGCACTATTT
4605 SEQ ID NO:3909 -11.4 -29.9 76.8 -18.5 0 -6.1
TCAAAAAGCATTCAAAGAAA
4703 SEQ ID NO:3910 -11.4 -26.2 71.8 -14.8 0 -27
AAAAGCAAATGATTGTTTTT
4767 SEQ ID NO:3911 -11.4 -23.5 67.8 -8.6 -3.5 -9.8
GTGAGGGTGAAGACGCAGAA
255 SEQ ID NO: 3912 -11.3 -38.9 89.1 -26.4 -1.1 -4.5
CCAGATCTCCATTATCCTTA
903 SEQ ID NO: 3913 -11.3 -33.6 81.8 -22.3 0 -6.6
TCTTCGAATTTTATCGATGA
1523 SEQ ID NO: 3914 -11.3 -28.4 73.4 -13.7 -3.4 -9.3
CCAGGTTCATTCCAGCCTGA
1578 SEQ ID NO: 3915 -11.3 -39.5 90 -21.5 -67 -15.9
CCAGGGTAGGGGTGAGTTGT
1710 SEQ ID NO: 3916 -11.3 -41.7 94.3 -29.5 -0.7 -4.4
GGTTCAATAACCTCCAACAG
1735 SEQ ID NO: 3917 -11.3 -33.7 83.4 -20.4 -2 -7.4
GAGCAAAACACAGCAGGTTT
1986 SEQ ID NO:3918 -11.3 -34.2 84.6 -21.6 -1.2 -4.2
TTGGAAGCAATAGTTAAGGA
2327 SEQ ID NO:3919 -11.3 -31.6 80.9 -19.6 -0.4 -2.7
CAATAAAAGCTATTAATTCG
2501 SEQ ID NO: 3920 -11.3 -23.9 69.1 -12.6 0 -5.9
TCTGGGATATATTAACTAAT
2695 SEQ ID NO: 3921 -11.3 -28.1 74.8 -16.8 0 -5.7
GGAATCACAACTTTTAAGAA
3490 SEQ ID NO:3922 -11.3 -27.9 74.8 -16.6 0 -4.2
TACCATCAGGAAAGATTAAC
3651 SEQ ID NO: 3923 -11.3 -30.9 78.9 -18.7 -0.8 -2.9
GGTGCACACAGAAAGGGCTA
3829 SEQ ID NO: 3924 -11.3 -39.4 91.3 -26.6 0 -11 <cal/mol kcal/mol degC kcal/mol kcal/mol kcal/mol Intra Inter total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
ATATATAACATGTCATGATA
4142 SEQ ID NO: 3925 -11.3 -26.2 70.6 -13.7 0 -10.4
TATAAATCATATAGGTTATC
4191 SEQ ID NO:3926 -11.3 -25.1 70 -13.3 -0.2 -5.3
TTATTAAATAATTTCTCCAA
4747 SEQ ID NO: 3927 -11.3 -22.6 66.1 -11.3 0 -2.8
CCAAAAGCTTTAAGTCTGTT
480 SEQ ID NO: 3928 -11.2 -29.6 787 -17.8 0 -8.6
TCGTCTCTTTACCTGGGGAC
738 SEQ ID NO: 3929 -11.2 -39.2 90.1 -22 -6 -14.4
GGACTCTCATTCGTCTCTTT
748 SEQ ID NO: 3930 -11.2 -347 82.7 -21.4 -2.1 -5.5
GTCTGATCTCCAAGGACTCT
761 SEQ ID NO: 3931 -11.2 -38 86.7 -24.5 -2.3 -9.4
TCTGATCCTGCTGTTGAGAA
1155 SEQ ID NO: 3932 -11.2 -36.5 85.3 -25.3 0 -3.4
ATCGATGATGCAATCATTCC
1511 SEQ ID NO: 3933 -11.2 -32.7 78.7 -19.3 -1.7 -12.4
TTTTTCTTCGAATTTTATCG
1527 SEQ ID NO: 3934 -11.2 -24 68 -12.8 0 -5.9
ATTGTCTATATGATCTCCAC
1959 SEQ ID NO:3935 -11.2 -31.6 78.3 -20.4 0 -4.2
ACTTGATTGTCTATATGATC
1964 SEQ ID NO: 3936 -11.2 -29.2 74.9 -17.3 -0.4 -5.8
AAATGTTTGGAAGCAATAGT
2333 SEQ ID NO:3937 -11.2 -28.8 76.4 -16.8 -0.6 -8
TTGATGAAACAGAAGTTTTT
2441 SEQ ID NO: 3938 -11.2 -26.3 71.9 -14.2 -0.8 -4.3
CTGGGATATATTAACTAATA
2694 SEQ ID NO: 3939 -11.2 -27 73.8 -15.8 0 -6.1
AGAAAGTCTAGAAAATTTCA
2838 SEQ ID NO: 3940 -11.2 -26.9 727 -14.6 -0.7 -9.8
GATATACACTTGTAAACTTT
2923 SEQ ID NO: 3941 -11.2 -26.1 72.6 -14.3 -0.3 -6.3
TCTGACATACAGTTCTAAAT
3699 SEQ ID NO: 3942 -11.2 -29.8 77 -17.9 -0.4 -3.2
AATCATTTTAATAAATTGCA
4069 SEQ ID NO: 3943 -11.2 -22.2 65.1 -11 0 -3.2
AAAAAATATATAACATGTCA
4147 SEQ ID NO: 3944 -11.2 -22.7 66 -11.5 0 -6.2
TGCTTTGGACAGATCTGGCT
356 SEQ ID NO: 3945 -11.1 -38.4 88.6 -24.5 -1.9 -13.6
CTGCTTTGGACAGATCTGGC
357 SEQ ID NO:3946 -11.1 -38.4 88.3 -24.6 -07 -13.6
TCCCAGGTCATTTCCCATCA
422 SEQ ID NO:3947 -11.1 -387 88.5 -27.1 -0.2 -3.3
CTTCCAAAAGCTTTAAGTCT
483 SEQ ID NO: 3948 -11.1 -29.8 78.6 -18.1 0 -8.6
ACTGTTCGACCAGGGAAGTT
1276 SEQ ID NO: 3949 -11.1 -37.4 88.2 -23.9 -1.6 -12.7
TCATGGTCTTATCCAAAAAT
2349 SEQ ID NO:3950 -11.1 -29.4 76.1 -16.2 -2.1 -6.8 kcal/mol kcal/mol degC kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecular Site oligo binding formation Duplex structure oligo oligo
ACAATAAAAGCTATTAATTC
2502 SEQ ID NO:3951 -11/ -237 68.6 -12.6 0 -5.9
TTTCCTGTACCATCAGGAAA
3658 SEQ ID NO: 3952 -11.1 -33.9 83.4 -15 -7.8 -18
CATACAGTTCTAAATCACAA
3694 SEQ ID NO: 3953 -11/ -28.3 75.3 -17.2 0 -1.6
CAAACATGGATGTCTGACAT
3711 SEQ ID NO: 3954 -11/ -32.8 80.2 -20.2 -0.5 -10.9
GTGTAGTTTTACAAACATGG
3722 SEQ ID NO:3955 -11/ -28.9 77.6 -16.9 -0.6 -9
TTACACAAATAATTTGGCCA
3920 SEQ ID NO: 3956 -11.1 -28.9 77.3 -16.9 0 -9.5
AGATTTTCAGTTTTCTATTA
3937 SEQ ID NO:3957 -11.1 -24.8 69.5 -13.7 0 -2.1
AGTTCCATATTTTTAATATT
3957 SEQ ID NO:3958 -11/ -22.6 66.1 -11.5 0 -3.2
CAGTAGCCCTTCCCTTCCCA
4039 SEQ ID NO: 3959 -11/ -41.8 94.3 -30.7 0 -1.2
GCTGCCTGTATGAATGCATG
4218 SEQ ID NO:3960 -11.1 -36.3 85.6 -24.2 -0.5 -9.6
AGCAAATGATTGTTTTTTTA
4764 SEQ ID NO:3961 -11.1 -23.9 68.6 -12.1 -0.4 -6.1
GCTTTGGACAGATCTGGCTG
355 SEQ ID NO: 3962 -11 -38.4 88.3 -24.6 -1.9 -13.7
GGAATCCCAGGTCATTTCCC
426 SEQ ID NO:3963 -11 -38.7 88.4 -25.8 -1.9 -10.2
TTTTTGGAAACTCCTTCTCT
588 SEQ ID NO: 3964 -11 -31.1 79.5 -19 -1 -6.3
ACAGGTCTGATCTCCAAGGA
765 SEQ ID NO:3965 -11 -38.9 88.4 -26.3 -1.1 -11.1
CACACCATGAACAGAAATGG
1085 SEQ ID NO: 3966 -11 -33.5 82.1 -197 -2.8 -7.7
TTGCCGCCCTCCTAACATGT
1823 SEQ ID NO: 3967 -11 -39.8 91.8 -28.8 0 -5.2
CTAATTTCATCAAATAGCTC
2182 SEQ ID NO: 3968 -11 -27.4 73.5 -16.4 0 -3.8
ATTAAGGCAGTCACTTTTGA
2457 SEQ ID NO: 3969 -11 -31.3 80.1 -20.3 0 -6
TATTAAGGCAGTCACTTTTG
2458 SEQ ID NO:3970 -11 -30.2 79.2 -19.2 0 -6.2
TCACACAGATGATTGATTAA
3116 SEQ ID NO:3971 -11 -29.8 75.9 -18.8 0 -7
CAACAGTGAAGAAATTCACA
3778 SEQ ID NO: 3972 -11 -30 77.3 -12.2 -6.8 -13.8
TAAGGTGCACACAGAAAGGG
3832 SEQ ID NO:3973 -11 -36.9 88.4 -24.4 0 -11
ATAAAACAATCTCATCTAAA
4125 SEQ ID NO: 3974 -11 -25 69.4 -14 0 0
CTCCCTCTTCCCCTAGAGCA
4262 SEQ ID NO: 3975 -11 -42.3 93.8 -28.6 -27 -7.8
TTGGTGGAATGACATTAAAA
1182 SEQ ID NO: 3976 -10.J ) -29 76.3 -14.9 -3.2 -11.2 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GCAATCATTCCTTCCAGCAC
1502 SEQ ID NO: 3977 -10.9 -36.2 85.3 -25.3 0 -2.9
TGCAATCATTCCTTCCAGCA
1503 SEQ ID NO: 3978 -10.9 -36.1 85.4 -25.2 0 -4.1
CAGGGTAGGGGTGAGTTGTG
1709 SEQ ID NO: 3979 -10.9 -40.5 92.5 -29.6 0 -2.7
TCCAAGTTGAGTCTGGAACA
1785 SEQ ID NO: 3980 -10.9 -36.1 85.8 -22.4 -2.5 -13.3
AAAATGTTTGGAAGCAATAG
2334 SEQ ID NO:3981 -10.9 -27.5 74.6 -15.8 -0.6 -8
CACCAACAGAAAGTCTAGAA
2845 SEQ ID NO: 3982 -10.9 -32.6 81.8 -21.2 0 -7.5
CTGATATACACTTGTAAACT
2925 SEQ ID NO: 3983 , -10.9 -28.5 76.2 -17 -0.3 -6.3
CACAACTTCCCTTTTCTGAT
2940 SEQ ID NO: 3984 -10.9 -32.3 80.9 -21.4 0 -1.4
CACACAGATGATTGATTAAT
3115 SEQ ID NO: 3985 -10.9 -28.5 74.1 -17.6 0 -7
ATGAGCTTTTTTTTTTTTTT
3222 SEQ ID NO:3986 -10.9 -20.8 63.4 -9.9 0 -6.4
AATCACAACTTTTAAGAAGT
3488 SEQ ID NO: 3987 -10.9 -26.5 72.8 -15.1 -0.1 -8
TTCCATATTTTTAATATTAG
3955 SEQ ID NO:3988 -10.9 -21.7 65 -10.8 0 -4.6
ACCTTCACTGCACACAGGAC
4292 SEQ ID NO: 3989 -10.9 -39.4 90.3 -267 -1.8 -8.8
TAAACTAAACCTGCGCTGAC
4399 SEQ ID NO: 3990 -10.9 -34.1 85 -22.7 0 -7.5
GTAAACTAAACCTGCGCTGA
4400 SEQ ID NO: 3991 -10.9 -34.1 85 -22.7 0 -7.5
AGGAGGGAAATATATTTTTT
12 SEQ ID NO: 3992 -10.8 -27 73.5 -15.3 0 -9.6
CTCTTTACCTGGGGACCCAG
734 SEQ ID NO: 3993 -10.8 -40.6 92.6 -23.5 -5.1 -20.7
TCAGAGTCCCCAGAGAAGTC
1257 SEQ ID NO: 3994 -10.8 -40.2 89.9 -29.4 0 -3.1
CATGCTGGGCAGTTTTTTCT
1540 SEQ ID NO:3995 -10.8 -34.2 84.3 -21 -0.4 -12.9
AGCTTCCAGGTTCATTCCAG
1583 SEQ ID NO: 3996 -10.8 -37.1 87.2 -24.9 -1.3 -4.9
TCTGGAACAGAGCTATCATA
1774 SEQ ID NO: 3997 -10.8 -34.8 83.5 -22.6 -1.2 -9.9
CCTCCTAACATGTTGAGCGT
1816 SEQ ID NO: 3998 -10.8 -36.5 86.9 -24.1 0 -11.3
ATTGGTGATGATTTCAGCTA
2390 SEQ ID NO:3999 -10.8 -31.9 79.5 -19.8 -1.2 -7.3
AGAAGTTTTTTGATATTTCC
2431 SEQ ID NO:4000 -10.8 -25.6 71 -13.2 -1.5 -7
TTCGACTTTCTTTAAGGCAA
2485 SEQ ID NO:4001 -10.8 -30.4 79.1 -19.6 0 -5.4
ATAAAAGCTATTAATTCGAC
2499 SEQ ID NO: 4002 -10.8 -25.5 71.1 -14.7 0 -5.9
kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
TAGTTCACTAAATATAAAGG
2996 SEQ ID NO:4003 -10.8 -26.3 73.3 -15.5 0 -4.9
AGAAAAAATGAGCAAGCGTA
3014 SEQ ID NO:4004 -10.8 -30.4 78.9 -19.6 0 -4.8
TTAGAAGCACCAACCCATTT
3136 SEQ ID NO:4005 -10.8 -33.4 83.7 -22.6 0 -2.7
AGGAAAGATTAACCAATTGG
3644 SEQ ID NO:4006 -10.8 -30.1 78.3 -18 0 -10.6
TTTACAAACATGGATGTCTG
3715 SEQ ID NO:4007 -10.8 -30.3 78 -18.5 0 -9.9
ACAGTGAAGAAATTCACAGG
3776 SEQ ID NO:4008 -10.8 -32.4 80.8 -14.6 -7 -14
GATTTTCAGTTTTCTATTAC
3936 SEQ ID NO:4009 -10.8 -24.9 70.1 -14.1 0 -2.1
TAAATCATATAGGTTATCCA
4189 SEQ ID NO:4010 -10.8 -28.1 747 -16.1 -1.1 -5.3
TGCACTATTTACATATTGCT
4595 SEQ ID NO:4011 -10.8 -29.6 77.7 -16.9 -1.9 -8.6
CATCACATCTCCCCTCTCCT
194 SEQ ID NO:4012 -107 -40.2 89.4 -29.5 0 0
TTTTGGAAAATCAACCAAAA
314 SEQ ID NO:4013 -10.7 -25.9 71.9 -11.6 -3.6 -9.1
GTTCAGAGTCCCCAGAGAAG
1259 SEQ ID NO:4014 -107 -387 88.5 -28 0 -2.9
ACATCCAGGAGTACTGCAGT
1917 SEQ ID NO: 4015 -10.7 -38.7 89.4 -26.2 -1.4 -11.5
AAACATACAGCATGTGTTTA
2061 SEQ ID NO:4016 -107 -29.1 77.2 -15.2 -3.2 -10.9
ACACAGACTTTGGGCACTGG
2777 SEQ ID NO:4017 -10.7 -39 90.5 -27.5 -0.6 -7.3
CCCTTTTCTGATATACACTT
2932 SEQ ID NO:4018 -10.7 -30.6 78.9 -19.9 0 -1.6
CCCATCTTAAACAGCTGTAC
3059 SEQ ID NO:4019 -10.7 -33.9 84.1 -20.5 0 -13.6
CAGATATATACAAAATATGA
3238 SEQ ID NO:4020 -10.7 -247 69 -14 0 -3.5
ACAAATAGTTTACCAGCTTT
3438 SEQ ID NO:4021 -10.7 -28.8 77.6 -18.1 0 -4
CTAAATCACAAAAATCAGTA
3685 SEQ ID NO: 4022 -10.7 -25.8 71.6 -15.1 0 -0.8
TAATTTTCAACAGTGAAGAA
3785 SEQ ID NO:4023 -10.7 -26.7 72.7 -13.4 -2.6 -10.4
AGTAAACTAAACCTGCGCTG
4401 SEQ ID NO:4024 -10.7 -33.8 85.1 -22.6 0 -7.5
AGAAAACAAAAACATGTCCT
4688 SEQ ID NO:4025 -10.7 -28.5 75.8 -17.8 0 -6.2
ATAACAAATAACATTCAAAA
4717 SEQ ID NO:4026 -107 -22.1 65.2 -11.4 0 0
AGTGAAACTGCTTTGGACAG
364 SEQ ID NO: 027 -10.6 -34.4 84.4 -23.3 0 -7.6
TTCTCTGTGGGGGCAGCAGA
574 SEQ ID NO:4028 -10.6 -43 94.7 -28.2 -4.2 -11.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
AAACTCCAAATCCTGCAAAA
710 SEQ ID NO:4029 -10.6 -30.8 79.4 -20.2 0 -4.9
GCAGTCCTCATTCGAGTTTC
848 SEQ ID NO:4030 -10.6 -35.7 84.3 -23.7 -1.3 -7.6
AACATCCAGGAGTACTGCAG
1918 SEQ ID NO:4031 -10.6 -37.4 87.8 -25.1 -1.7 -8.7
AGCAAAACACAGCAGGTTTG
1985 SEQ ID NO: 4032 -10.6 -33.9 84.6 -22 -1.2 -6.5
AATAAATTTCACCATCTACT
2678 SEQ ID NO: 4033 -10.6 -27.2 73.3 -16.6 0 -0.9
ACAACTTCCCTTTTCTGATA
2939 SEQ ID NO:4034 -10.6 -31.5 80.1 -20.9 0 -1.4
GGGTGTCTAGCCATTTTTGC
3602 SEQ ID NO:4035 -10.6 -35.9 86.6 -22.6 -2.7 -8.9
ATAATTTTCAACAGTGAAGA
3786 SEQ ID NO:4036 -10.6 -26.9 72.6 -14.5 -1.8 -8.6
TTAGATTTTCAGTTTTCTAT
3939 SEQ ID NO:4037 -10.6 -24.8 69.5 -13.3 -07 -5.3
AAAATCATTCTTATTTTACA
4107 SEQ ID NO:4038 -10.6 -22.3 65.2 -11.7 0 -0.4
CTAAAAATCATTCTTATTTT
4110 SEQ ID NO: 4039 -10.6 -21 63.3 -10.4 0 -0.2
CAAACTTCTGAAGCTTCTGT
4552 SEQ ID NO: 4040 -10.6 -32.2 81 -19.4 0 -12.5
CAGGAGGGAAATATATTTTT
13 SEQ ID NO: 4041 -10.5 -28.2 75.3 -17 0 -8.8
CAACTTAGCTTGTGAACGCA
40 SEQ ID NO:4042 -10.5 -337 84 -22 -1.1 -6.9
TCAGTGAATATCAACTCTGG
114 SEQ ID NO:4043 -10.5 -33.2 81.1 -21.9 -0.6 -5.9
GAAACTGCTTTGGACAGATC
361 SEQ ID NO: 4044 -10.5 -33.9 82.4 -22.2 -1 -9.3
GAGAGGCTTGCAGTCCTCAT
857 SEQ ID NO: 4045 -10.5 -39.9 89.8 -22.7 -5.8 -21.5
AATTGGTGGAATGACATTAA
1184 SEQ ID NO: 4046 -10.5 -29.2 76.2 -16 -2.7 -10.2
TTCAATAACCTCCAACAGTG
1733 SEQ ID NO:4047 -10.5 -32.5 81.7 -22 0 -1.5
ATAGCTCTTGGCTCTTCAGA
2169 SEQ ID NO:4048 -10.5 -36.6 86.2 -24.5 -1.5 -8
TCACTGAAAAGTGATGACGA
2652 SEQ ID NO: 4049 -10.5 -33.3 81.1 -16.6 -6.2 -13.4
AAGTTCATCACACAGACTTT
2786 SEQ ID NO: 4050 -10.5 -31.3 79.1 -20.8 0 -3.6
CCAACTGTGAAAAAAAGTAT
2813 SEQ ID NO: 4051 -10.5 -27.4 74.8 -16.9 0 -3.7
TCCCTTTTCTGATATACACT
2933 SEQ ID NO:4052 -10.5 -32.1 80.6 -21.6 0 -1.6
AAGAAAAAATGAGCAAGCGT
3015 SEQ ID NO:4053 -10.5 -30 78.1 -19.5 0 -4.7
AAAAATTATGTAAGAAAAAA
3026 SEQ ID NO:4054 -10.5 -19.4 607 -8.9 0 -3.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CTACGAACTAGCTGCCCATC
3073 SEQ ID NO:4055 -10.5 -38.1 88.9 -27.6 0 -6.3
GTTATCTGGAATCACAACTT
3497 SEQ ID NO:4056 -10.5 -30.6 78.1 -20.1 0 -6.3
GGTAAGGTGCACACAGAAAG
3834 SEQ ID NO:4057 -10.5 -35.8 86.7 -23.8 0 -11
TCTCCCTCTTCCCCTAGAGC
4263 SEQ ID NO:4058 -10.5 -42.6 937 -29.7 -2.4 -7.5
ATTCAACTGCTGGGGGAGGG
4528 SEQ ID NO:4059 -10.5 -41.6 93.2 -29.6 -1.4 -9.1
GCAAACTATTGCCAAACTTC
4564 SEQ ID NO:4060 -10.5 -31.1 80.3 -18.7 -1.9 -6.6
CAGTCCTCATTCGAGTTTCC
847 SEQ ID NO:4061 -10.4 -35.6 84.3 -24.1 -1 -7.6
ATGCAATCATTCCTTCCAGC
1504 SEQ ID NO: 4062 -10.4 -35.1 83.6 -24.7 0 -4.9
GCCCTCCTAACATGTTGAGC
1818 SEQ ID NO: 4063 -10.4 -38.6 89.3 -26.6 0 -11.3
CAGGAGCAAAACACAGCAGG
1989 SEQ ID NO:4064 -10.4 -377 88.8 -26.2 -1 -2.9
CATAAGATACCTGAAGCCTG
2094 SEQ ID NO:4065 -10.4 -34.2 83.6 -23.8 0 -1.3
CATCAAATAGCTCTTGGCTC
2175 SEQ ID NO: 4066 -10.4 -34.1 82.8 -22.1 -1.5 -8
GTGGTCTTCTGATAGCTAAG
3166 SEQ ID NO:4067 -10.4 -34.4 83.8 -21.8 -2.2 -10.4
ATCACAACTTTTAAGAAGTT
3487 SEQ ID NO:4068 -10.4 -26.5 72.8 -15.1 -0.7 -9.5
CCATATTTTTAATATTAGAT
3953 SEQ ID NO:4069 -10.4 -21.9 65 -10.8 0 -8.7
GAGGCTTGCAGTCCTCATTC
855 SEQ ID NO:4070 -10.3 -38.7 88.3 -217 -5.2 -21.5
CATTAAAAATAGGCTTCTGA
1170 SEQ ID NO:4071 -10.3 -28.1 75.4 -17.8 0 -5
AATGTTTGGAAGCAATAGTT
2332 SEQ ID NO: 4072 -10.3 -28.8 76.4 -17.8 -0.5 -8
GATGATTTCAGCTAACATCT
2384 SEQ ID NO:4073 -10.3 -31 77.8 -19.1 -1.5 -8.3
CAGCTGTGTTACAGCTGGTT
3514 SEQ ID NO:4074 -10.3 -36.8 88.3 -16.9 -9.4 -26.8
AATGTGAAAATGGGTGTCTA
3613 SEQ ID NO:4075 -10.3 -31.5 79.9 -21.2 0 -6.8
TGTTTAAAATAATTTTCAAC
3794 SEQ ID NO:4076 -10.3 -20.7 63.5 -10.4 0 -4.6
TTTGACCCCTACAAAAAATA
4159 SEQ ID NO:4077 -10.3 -28.9 77.6 -18.6 0 -1.7
TCTATGCAAACTATTGCCAA
4569 SEQ ID NO:4078 -10.3 -31.6 80.6 -19.4 -1.9 -6.6
TGAGCAAGCACACTGCTGGG
175 SEQ ID NO:4079 -10.2 -41.4 93.1 -28.3 -2.9 -9.3
TGAGAGGCTTGCAGTCCTCA
858 SEQ ID NO -.4080 -10.2 -40.9 91.6 -23.9 -6.2 -21.5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
GGTGGAATGACATTAAAAAT
1180 SEQ ID NO:4081 -10.2 -28 74.6 -16.5 -1.2 -7.2
TAGTTTATACAATAAAAGCT
2510 SEQ ID NO:4082 -10.2 -24.9 71.3 -147 0 -3.3
GAAAACAATAAAAAATAAAA
2557 SEQ ID NO:4083 -10.2 -18.2 58.8 -8 0 0
GAGCAAGCGTAGTTCACTAA
3005 SEQ ID NO: 4084 -10.2 -34.4 84.8 -21.4 -2.8 -11.3
AACTTTTAAGAAGTTATACA
3482 SEQ ID NO:4085 -10.2 -24.6 70.6 -12.8 -1.4 -10.6
AATCAGTAGCTGAGCTTTCC
3673 SEQ ID NO: 4086 -10.2 -35.2 84.8 -22.2 -1.5 -13.8
TAAATCACAAAAATCAGTAG
3684 SEQ ID NO:4087 -10.2 -25.8 71.6 -15.6 0 -0.8
TTTAAAATAATTTTCAACAG
3792 SEQ ID NO:4088 -10.2 -20.6 63.3 -10.4 0 -2.1
CACACAGGACCAGTCTCCAT
4282 SEQ ID NO:4089 -10.2 -39.7 89.8 -27.1 -2.4 -8.8
AAACTAAACCTGCGCTGACA
4398 SEQ ID NO: 4090 -10.2 -34.9 85.9 -24.2 0 -7.5
ATTCTGCACTATTTACATAT
4599 SEQ ID NO: 4091 -10.2 -27.6 73.9 -17.4 0 -4.3
CTTCAAATGTTGCTGTTCTG
626 SEQ ID NO: 4092 -10.1 -30.9 79.1 -20.8 0 -1.7
GGACCCAGAAGAAAACTCCA
722 SEQ ID NO:4093 -10.1 -37.2 87.5 -26.2 -0.8 -5.8
TGCAGTCCTCATTCGAGTTT
849 SEQ ID NO: 4094 -10.1 -35.4 84.5 -23.9 -1.3 -7.6
GCAGTGTTACATTACTGGGG
933 SEQ ID NO:4095 -10.1 -36.1 87.3 -21.4 -4.6 -10.9
ATTGGTGGAATGACATTAAA
1183 SEQ ID NO: 4096 -10.1 -29.2 76.2 -15.6 -3.5 -12
GAATAGCCATTAGAAAAAAC
1294 SEQ ID NO:4097 -10.1 -27 73.9 -16.9 0 -2.8
TGACACCAGGGTAGGGGTGA
1715 SEQ ID NO:4098 -10.1 -43.2 95.7 -28.1 -5 -13
GTTCAATAACCTCCAACAGT
1734 SEQ ID NO:4099 -10.1 -32.6 81.8 -22.5 0 -1.3
GCTTTTCCTAGCTCTTTGAT
2212 SEQ ID NO: 4100 -10.1 -32.7 81.7 -20.5 -2.1 -6.6
AGTTTTGTCAGTTGATAAAA
2275 SEQ ID NO:4101 -10.1 -26.5 72.8 -12.8 -3.4 -14.6
TTTGATGAAACAGAAGTTTT
2442 SEQ ID NO:4102 -10.1 -26.3 71.9 -15.2 -0.9 -3.7
AATAAAAGCTATTAATTCGA
2500 SEQ ID NO:4103 -10.1 -24.2 68.9 -14.1 0 -5.9
TCTGATATACACTTGTAAAC
2926 SEQ ID NO: 4104 -10.1 -28.8 76.2 -18.1 -0.3 -6.3
CTAAATATAAAGGAAATTGT
2989 SEQ ID NO: 4105 -10.1 -23.6 68.4 -13.5 0 -1
AAAATTATGTAAGAAAAAAT
3025 SEQ ID NO:4106 -10.1 -19.6 60.7 -9.5 0 -4.2 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TTCACACAGATGATTGATTA
3117 SEQ ID NO: 4107 -10.1 -29.8 75.9 -19.7 0 -5.2
GGTTATCTGGAATCACAACT
3498 SEQ ID NO:4108 -10.1 -33 81.5 -22 -0.7 -6.5
TCAGCTGTGTTACAGCTGGT
3515 SEQ ID NO:4109 -10.1 -38.3 89.8 -18.3 -97 -27.4
ATTACACAAATAATTTGGCC
3921 SEQ ID NO:4110 -10.1 -27.9 75.5 -17.3 0 -7.6
CCTGTATGAATGCATGTACA
4214 SEQ ID NO:4111 -10.1 -33.1 81.8 -21.1 -1.7 -11.3
AATAACAAATAACATTCAAA
4718 SEQ ID NO: 4112 -10.1 -22.1 65.2 -12 0 0
TTTATTAAATAATTTCTCCA
4748 SEQ ID NO:4113 -10.1 -22.6 66.1 -12.5 0 -37
GCAAATGATTGTTTTTTTAT
4763 SEQ ID NO:4114 -10.1 -22.9 66.8 -12.8 0 -6.1
AGGGAAATATATTTTTTTTT
9 SEQ ID N0:4115 -10 -21.9 65.2 -10.7 0 -10.4
GAGCAAGCACACTGCTGGGG
174 SEQ ID NO:4116 -10 -42.6 94.3 -29.8 -2.8 -9
CTGTTTCCCCCGAGGAAAGG
465 SEQ ID NO: 4117 -10 -39.5 91 -26.1 -3.4 -11.2
TCTGTTTCCCCCGAGGAAAG
466 SEQ ID NO:4118 -10 -38.6 89.6 -25.2 -3.4 -11.2
TTGCAGTCCTCATTCGAGTT
850 SEQ ID NO:4119 -10 -35.4 84.5 -24 -1.3 -7.6
TCCAGGTTCATTCCAGCCTG
1579 SEQ ID NO:4120 -10 -39.5 90 -23.9 -5.6 -13.6
AAATGCCATAAGAAACATCC
1931 SEQ ID NO:4121 -10 -30.4 78 -20.4 0 -3.4
GCAAATGCCATAAGAAACAT
1933 SEQ ID NO: 4122 -10 -30.2 78 -19.5 -0.4 -5.7
CAAAACACAGCAGGTTTGCA
1983 SEQ ID NO:4123 -10 -33.9 84.5 -21.6 -1.7 -12.5
CACTGAAAAGTGATGACGAC
2651 SEQ ID NO: 4124 -10 -33.1 81.1 -18.3 -4.8 -10.6
TAATAAATTTCACCATCTAC
2679 SEQ ID NO:4125 -10 -26.4 72.4 -16.4 0 -0.9
AAAGTTCATCACACAGACTT
2787 SEQ ID NO:4126 -10 -31.3 79.1 -20.8 -0.1 -4
GCACAACTTCCCTTTTCTGA
2941 SEQ ID NO:4127 -10 -34.6 84.4 -24.6 0 -2.7
CTATAAAAGGCACAACTTCC
2950 SEQ ID NO:4128 -10 -31.4 81 -21.4 0 -2.4
TATGAGCTTTTTTTTTTTTT
3223 SEQ ID NO:4129 -10 -21.2 64.3 -11.2 0 -6.4
GAATCACAACTTTTAAGAAG
3489 SEQ ID NO:4130 -10 -267 73 -16.7 0 -5
GTAAGGTGCACACAGAAAGG
3833 SEQ ID NO:4131 -10 -35.8 86.7 -24.3 0 -11
AGTAACCCGCTATTAGATGG
4497 SEQ ID NO:4132 -10 -347 85.3 -24 0 -8.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TTCTGCACTATTTACATATT
4598 SEQ ID NO:4133 -10 -27.4 74.1 -17.4 0 -4.3
TTTTTATTAAATAATTTCTC
4750 SEQ ID NO:4134 -10 -19 59.9 -9 0 -3.7
GCCATTAGAAAAAACTGTTC
1289 SEQ ID NO:4135 -9.9 -28.9 76.8 -19 0 -2.3
TGGGCAGTTTTTTCTTCGAA
1535 SEQ ID NO:4136 -9.9 -32.4 81.8 -22 -0.2 -5.9
AGTGACACCAGGGTAGGGGT
1717 SEQ ID NO:4137 -9.9 -43 95.9 -31.8 -1.2 -8.9
GGAGCAAAACACAGCAGGTT
1987 SEQ ID NO:4138 -9.9 -36.6 87.7 -25.4 -1.2 -3.1
GCATAGAATCCAAGAGTTTT
2289 SEQ ID NO: 4139 -9.9 -30.5 78.2 -20.6 0 -2.7
AAGCTATTAATTCGACTTTC
2495 SEQ ID NO: 4140 -9.9 -27.6 74.2 -17.7 0 -5
AAAAAATAAAACAACAAAAC
2548 SEQ ID N0:4141 -9.9 -19.9 62.6 -10 0 0
GTTTCCATGCATAAATCAAA
2896 SEQ ID NO:4142 -9.9 -28 74.4 -18.1 0 -7
CTGAAAGACAAATAGTTTAC
3445 SEQ ID NO:4143 -9.9 -26.9 73.9 -16.5 -0.2 -2.8
CAACTTTTAAGAAGTTATAC
3483 SEQ ID NO: 4144 -9.9 -24.6 71 -12.8 -1.7 -11.5
ATCTCATCTAAAAATCATTC
4117 SEQ ID NO: 4145 -9.9 -26.5 70.6 -16.6 0 0
TAAAACAATCTCATCTAAAA
4124 SEQ ID NO: 4146 -9.9 -24.8 69.5 -14.9 0 0
CAAAAAATATATAACATGTC
4148 SEQ ID N0:4147 -9.9 -227 66.6 -12.8 0 -6.2
CAAAAAGCATTCAAAGAAAA
4702 SEQ ID NO: 4148 -9.9 -24.7 70 -14.8 0 -2.7
TGTGAACGCAGAAGGAGCAG
30 SEQ ID NO: 4149 -9.8 -38.6 89.1 -27.7 -1 -7
ACTGCTTTGGACAGATCTGG
358 SEQ ID NO:4150 -9.8 -37.2 87 -247 -1 -13.6
AGAGTGAGTTTTTGGAAACT
596 SEQ ID NO: 4151 -9.8 -31 79.5 -18.6 -0.9 -13.4
GAAAACTCCAAATCCTGCAA
712 SEQ ID NO:4152 -9.8 -32.3 81 -22.5 0 -4.9
CCTCATTCGAGTTTCCTTCC
843 SEQ ID NO:4153 -9.8 -35.5 84.4 -24.3 -1.3 -7.6
GGAACTATTGTTTTGTTACC
1678 SEQ ID NO: 4154 -9.8 -29.1 77.7 -17.7 -1.5 -5.2
CTCCAACAGTGACACCAGGG
1724 SEQ ID NO:4155 -9.8 -40.4 91.6 -30.6 0 -7.3
CGCCCTCCTAACATGTTGAG
1819 SEQ ID NO:4156 -9.8 -37.6 88.2 -26.3 0 -10.9
GATTGTCTATATGATCTCCA
1960 SEQ ID NO:4157 -9.8 -31.8 78.2 -20.4 -1.6 -8.2
AATTTCATCAAATAGCTCTT
2180 SEQ ID NO:4158 -9.8 -27 72.4 -17.2 0 -3.8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
ATTCTAATTTCATCAAATAG
2185 SEQ ID NO:4159 ■9.8 -23.9 67.5 -14.1 0 -0.9
TGATAGTTTATACAATAAAA
2513 SEQ ID NO:4160 -9.8 -22.9 67 -13.1 0 -2.3
GTTTAAAATAATTTTCAACA
3793 SEQ ID NO:4161 -9.8 -20.7 63.5 -10.4 0 -8
CAATCTCATCTAAAAATCAT
4119 SEQ ID NO: 4162 -9.8 -26.2 70.5 -16.4 0 0
ACAATCTCATCTAAAAATCA
4120 SEQ ID NO:4163 -9.8 -27.3 72.4 -17.5 0 0
ACACTGCTGGGGTTTTCTTC
166 SEQ ID NO:4164 -9.7 -36.8 87.7 -26.6 -0.2 -47
AAAACTCCAAATCCTGCAAA
711 SEQ ID NO:4165 -9.7 -30.8 79.4 -21.1 0 -4.9
GTCTCTTTACCTGGGGACCC
736 SEQ ID NO: 4166 -9.7 -41 92.6 -24.8 -6.5 -15.4
CCGCCAGAGGAGAAAGCAAA
801 SEQ ID NO:4167 -9.7 -38.4 89.5 -27.4 -1.2 -5.1
ACACCATGAACAGAAATGGC
1084 SEQ ID NO:4168 -9.7 -34.8 83.9 -21.9 -3.2 -8.7
CACTTGATTGTCTATATGAT
1965 SEQ ID NO:4169 -9.7 -28.9 74.8 -19.2 0 -4.2
GGATATATTAACTAATAAAT
2691 SEQ ID NO:4170 -9.7 -22.4 66.1 -12.7 0 -6.1
CTTCTATTTTGTTTAAAATA
3803 SEQ ID NO:4171 -97 -21 64.2 -11.3 0 -6.7
ACACAGGACCAGTCTCCATC
4281 SEQ ID NO:4172 -97 -40 89.8 -28.1 -2.2 -8.5
ATGAGATTCTGCACTATTTA
4604 SEQ ID NO:4173 -9.7 -29.9 76.8 -19.7 0 -8
GCATTCAAAGAAAACAAAAA
4696 SEQ ID NO: 4174 -97 -24.8 70.2 -15.1 0 -27
AAACTCCTTCTCTGTGGGGG
581 SEQ ID NO:4175 -9.6 -39.3 90.6 -24.6 -5.1 -12
GAAACTCCTTCTCTGTGGGG
582 SEQ ID NO:4176 -9.6 -38.4 88.8 -25 -3.8 -9.1
AGGTCTGATCTCCAAGGACT
763 SEQ ID NO:4177 -9.6 -38.9 88.5 -26.3 -3 -10.8
AACAGGTCTGATCTCCAAGG
766 SEQ ID NO:4178 -9.6 -37.4 87 -26.2 -1.1 -11.1
ACCAGGGAAGTTCAGAGTCC
1268 SEQ ID NO:4179 -9.6 . -39.7 90.3 -29 -1 -6.2
CAGTTTTTTCTTCGAATTTT
1531 SEQ ID NO:4180 -9.6 -24.1 68.7 -14.5 0 -5.9
AAATTATGTAAGAAAAAATG
3024 SEQ ID NO: 4181 -9.6 -20.8 63.3 -11.2 0 -4.2
3217 SEQ ID NO:4182 -9.6 -15.4 54.1 -5.8 0- 0
ATTTTCAGTTTTCTATTACA
3935 SEQ ID NO:4183 -9.6 -24.6 69.5 -15 0 -0.7
GGGAGATTGAGTAAACTAAA
4410 SEQ ID NO:4184 -9.6 -30.7 79.2 -21.1 0 -3.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TGATTGTTTTTTTATTAAAT
4758 SEQ ID NO:4185 -9.6 -19.6 60.7 -10 0 -2
ATGATTGTTTTTTTATTAAA
4759 SEQ ID NO:4186 -9.6 -19.6 60.7 -10 0 -1.2
AATGATTGTTTTTTTATTAA
4760 SEQ ID NO:4187 -9.6 -19.6 60.7 -10 0 -3
GGAGGGAAATATATTTTTTT
11 SEQ ID NO:4188 -9.5 -25.8 71.7 -14.8 0 -11
CAGAGTGAGTTTTTGGAAAC
597 SEQ ID NO:4189 -9.5 -31 79.5 -20 -1.4 -9.8
ATGAGAGGCTTGCAGTCCTC
859 SEQ ID NO:4190 -9.5 -39.9 89.8 -25.2 -4.1 -18.5
GGGCTTGACAAAACCAGATC
916 SEQ ID NO:4191 -9.5 -36 85.7 -24.9 -0.8 -11.1
AGTTTATACAATAAAAGCTA
2509 SEQ ID NO: 4192 -9.5 -24.9 71.3 -14.7 -0.4 -5
AATAAAACAACAAAACCTCT
2544 SEQ ID NO:4193 -9.5 -26.2 72.9 -16.7 0 0
CTAATAAATTTCACCATCTA
2680 SEQ ID NO: 4194 -9.5 -26.3 72.3 -16.8 0 -0.7
AAAGGAAATTGTTAAAACCA
2981 SEQ ID NO: 4195 -9.5 -26.1 72.9 -16.6 0 -2.5
TTAATAAATTGCATGTAAAG
4062 SEQ ID NO:4196 -9.5 -23.4 68.3 -12.3 0 -11.4
AATCTCATCTAAAAATCATT
4118 SEQ ID NO:4197 -9.5 -25 68.5 -15.5 0 0
TTTTATTAAATAATTTCTCC
4749 SEQ ID NO:4198 -9.5 -21.4 64.4 -11.9 0 -3.7
AAACTGTTCGACCAGGGAAG
1278 SEQ ID NO:4199 -9.4 -36.1 86.5 -24.3 -1.6 -12.7
ATAAAACAACAAAACCTCTA
2543 SEQ ID NO:4200 -9.4 -26.6 737 -17.2 0 0
TAAGAAAAAATGAGCAAGCG
3016 SEQ ID NO:4201 -9.4 -29.1 77.1 -19.7 0 -4.7
GCACCAACCCATTTTCACAC
3130 SEQ ID NO: 4202 -9.4 -35.6 85.9 -26.2 0 -27
ATGTGAAAATGGGTGTCTAG
3612 SEQ ID NO:4203 -9.4 -327 81.4 -22.7 0 -8.6
GTTCCATATTTTTAATATTA
3956 SEQ ID NO: 4204 -9.4 -21.8 65.2 -12.4 0 -3.3
TATATAACATGTCATGATAA
4141 SEQ ID NO:4205 -9.4 -26 70.7 -14.6 0 -12.2
CCTACAAAAAATATATAACA
4152 SEQ ID NO:4206 -9.4 -23.8 69.2 -14.4 0 -3.4
CCTGTCTCCTGTTTACATAC
4450 SEQ ID NO:4207 -9.4 -34.2 84 -24.8 0 -4.4
AGATTCTGCACTATTTACAT
4601 SEQ ID NO:4208 -9.4 -29.7 76.9 -20.3 0 -67
GAGATTCTGCACTATTTACA
4602 SEQ ID NO:4209 -9.4 -31 78.8 -21.1 0 -8
TGAGATTCTGCACTATTTAC
4603 SEQ ID NO:4210 -9.4 -31 78.8 -21.1 0 -8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CATTCAAAAAGCATTCAAAG
4706 SEQ ID NO:4211 -94 -261 719 -167 0 -27
ACTCTGGCAGAGGAGCCGCT
101 SEQ ID NO:4212 -93 -446 963 -299 -45 -188
CAGTGAATATCAACTCTGGC
113 SEQ ID NO:4213 -93 -342 828 -242 -04 -59
AGTGAGTTTTTGGAAACTCC
594 SEQ ID NO: 4214 -93 -322 813 -17 -42 -20
CACACAGACTTTGGGCACTG
2778 SEQ ID NO: 4215 -93 -378 887 -285 0 -56
TAGAAGCACCAACCCATTTT
3135 SEQ ID NO:4216 -93 -334 837 -241 0 -25
ACAACTTTTAAGAAGTTATA
3484 SEQ ID NO:4217 -93 -246 706 -134 -17 -115
AATAGAAATCAAATTTCTTG
3895 SEQ ID NO: 4218 -93 -233 667 -127 -12 -78
ACTTAGCTTGTGAACGCAGA
38 SEQ ID NO:4219 -92 -352 857 -247 -12 -9
CCTGAGCAAGCACACTGCTG
177 SEQ ID NO-4220 -92 -402 914 -282 -28 -93
GACCCAGAAGAAAACTCCAA
721 SEQ ID NO:4221 -92 -348 844 -256 0 -03
GGGACCCAGAAGAAAACTCC
723 SEQ ID NO: 4222 -92 -384 89 -274 -06 -117
TCTTTACCTGGGGACCCAGA
733 SEQ ID NO: 4223 -92 -409 93 -243 -62 -23
AGGAGAAAGCAAACAGTTTT
794 SEQ ID NO: 4224 -92 -307 796 -209 -03 -3.3
AGTCCTCATTCGAGTTTCCT
846 SEQ ID NO: 4225 -92 -356 845 -25 -13 -76
CAGATCTCCATTATCCTTAA
902 SEQ ID NO:4226 -92 -312 785 -22 0 -65
CTGGGCAGTTTTTTCTTCGA
1536 SEQ ID NO: 4227 -92 -336 834 -237 -04 -5
CCGCCCTCCTAACATGTTGA
1820 SEQ ID NO:4228 -92 -388 90 -282 0 -10.8
ATGTTTGGAAGCAATAGTTA
2331 SEQ ID NO:4229 -92 -292 772 -192 -06 -8
CTCATGGTCTTATCCAAAAA
2350 SEQ ID NO:4230 -92 -304 781 -191 -21 -68
TTTTGATGAAACAGAAGTTT
2443 SEQ ID NO: 4231 -92 -263 719 -161 -09 -36
GTTTATACAATAAAAGCTAT
2508 SEQ ID NO-4232 -92 -239 694 -147 0 -5
CAGAAAGTCTAGAAAATTTC
2839 SEQ ID NO: 4233 -92 -269 73 -172 0 -75
TTTTTTTTTTTTTTTTTTGA
3216 SEQ ID NO:4234 -92 -157 531 -65 0 0
3218 SEQ ID NO: 4235 -92 -167 565 -75 0 -17
ATAAATCATATAGGTTATCC
4190 SEQ ID NO: 4236 -92 -271 73 -174 -02 -53 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target imolecular molecul Site oligo binding formation Duplex structure oligo oligo
CTGAGCAAGCACACTGCTGG
176 SEQ ID NO: 4237 -9.1 -40.2 91.4 -28.3 -2.8 -9.3
CAGGTCTGATCTCCAAGGAC
764 SEQ ID NO: 4238 -9.1 -38.9 88.1 -28.2 -1.1 -11.2
CAGAGGTACTCACACCATGA
1095 SEQ ID NO:4239 -9.1 -37.7 87.6 -26.3 -2.3 -8.5
GACCAGGGAAGTTCAGAGTC
1269 SEQ ID NO: 4240 -9.1 -38.8 88.5 -27.6 -2.1 -8.9
GTCTGGAACAGAGCTATCAT
1775 SEQ ID NO: 4241 -9.1 -357 84.3 -25 -1.4 -10.6
TTCATCACACAGACTTTGGG
2783 SEQ ID NO: 4242 -9.1 -34.8 84 -25.7 0 -5.6
AGCTTTTTTTTTTTTTTTTT
3219 SEQ ID NO:4243 -9.1 -17.9 58.5 -8.8 0 -2.7
AGATATATACAAAATATGAG
3237 SEQ ID NO: 4244 -9.1 -24.7 69 -15.6 0 -3.5
GCAGTAGCCCTTCCCTTCCC
4040 SEQ ID NO: 4245 -9.1 -43.1 95.4 -33.5 -0.1 -2.7
TAATAAATTGCATGTAAAGC
4061 SEQ ID NO: 4246 -9.1 -25.9 72.4 -15.2 0 -11.4
TACTTTACATACTTTAGTGC
4433 SEQ ID NO: 4247 -9.1 -28.5 77.4 -19.4 0 -6.3
GGCTTTTCCTAGCTCTTTGA
2213 SEQ ID NO: 4248 -9 -34.9 85.3 -23.3 -2.6 -7.6
GTGATGATTTCAGCTAACAT
2386 SEQ ID NO:4249 -9 -30.8 77.8 -21.1 -0.5 -7.7
TGGTGATGATTTCAGCTAAC
2388 SEQ ID NO:4250 -9 -33 81.3 -22.2 -1.8 -8.5
TCATCACACAGACTTTGGGC
2782 SEQ ID NO:4251 -9 -37.3 87 -28.3 0 -5.6
ATAAAAGGCACAACTTCCCT
2948 SEQ ID NO: 4252 -9 -33.4 83.8 -24.4 0 -2.9
AACTAGCTGCCCATCTTAAA
3068 SEQ ID NO: 4253 -9 -33.8 84.4 -24.8 0 -6.3
ATATGAGCTTTTTTTTTTTT
3224 SEQ ID NO: 4254 -9 -21.4 64.3 -12.4 0 -6.4
CAAATAGTTTACCAGCTTTC
3437 SEQ ID NO:4255 -9 -29 77.5 -20 0 -4
TTGTTTAAAATAATTTTCAA
3795 SEQ ID NO:4256 -9 -19.4 60.6 -10.4 0 -4.6
ACAGCTTCTATTTTGTTTAA
3807 SEQ ID NO:4257 -9 -26.6 73.7 -17.6 0 -4.1
TAGATTTTCAGTTTTCTATT
3938 SEQ ID NO:4258 -9 -24.8 69.5 -15.1 -0.5 -4.4
AAAACATGTCCTCATTTTAT
4680 SEQ ID NO: 4259 -9 -26.8 72.5 -17.8 0 -6.2
AAATGATTGTTTTTTTATTA
4761 SEQ ID NO: 4260 -9 -19.6 607 -10 -0.3 -4.8
ACAGCCAGTGAGGGTGAAGA
262 SEQ ID NO: 4261 -8.9 -40.7 91.9 -27.9 -3.9 -8.9
AATGAGAGGCTTGCAGTCCT
860 SEQ ID NO: 4262 -8.9 -38.4 88.7 -27 -1.6 -13.1 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CTTCATAAGATACCTGAAGC
2097 SEQ ID NO: 4263 -8.9 -32.1 80.5 -21.4 -1.8 -7.2
ATGGCTTTTCCTAGCTCTTT
2215 SEQ ID NO: 4264 -8.9 -33.6 83.6 -21.9 -2.8 -87
TGCATAAATCAAAAATGCTA
2889 SEQ ID NO:4265 -8.9 -26.9 73.2 -16.1 -1.9 -7
ATATAAAGGAAATTGTTAAA
2985 SEQ ID NO:4266 -8.9 -22.4 66.1 -13.5 0 -1
AATATAAAGGAAATTGTTAA
2986 SEQ ID NO:4267 -8.9 -22.4 66.1 -13.5 0 -1
AAATATAAAGGAAATTGTTA
2987 SEQ ID NO:4268 -8.9 -22.4 66.1 -13.5 0 -1
TAAATATAAAGGAAATTGTT
2988 SEQ ID NO: 4269 -8.9 -22.4 66.1 -13.5 0 -1
AGCTTCTATTTTGTTTAAAA
3805 SEQ ID NO: 4270 -8.9 -24.1 697 -15.2 0 -2.7
TGCAAACTATTGCCAAACTT
4565 SEQ ID NO:4271 -8.9 -30.8 80.3 -20 -1.9 -6.6
TCTGCACTATTTACATATTG
4597 SEQ ID NO:4272 -8.9 -28.6 75.9 -19.7 0 -4.3
GGGTTTTCTTCTCTACCAGG
157 SEQ ID NO: 4273 -8.8 -36.1 86.8 -257 -1.5 -67
ACCCAGAAGAAAACTCCAAA
720 SEQ ID NO: 4274 -8.8 -33.3 83 -24.5 0 -0.3
CACCAGGGTAGGGGTGAGTT
1712 SEQ ID NO: 4275 -8.8 -41.7 94.3 -28.4 -4.5 -12.5
CCAACAGTGACACCAGGGTA
1722 SEQ ID NO: 4276 -8.8 -39.4 91.3 -29.8 -0.4 -8.9
CTTGATTGTCTATATGATCT
1963 SEQ ID NO: 4277 -8.8 -29.1 74.8 -18.7 -1.6 -8.2
TGGCTTTTCCTAGCTCTTTG
2214 SEQ ID NO:4278 -8.8 -34.6 85.3 -23 -2.8 -8.7
TTATACAATAAAAGCTATTA
2506 SEQ ID NO: 4279 -8.8 -23 67.9 -14.2 0 -5
TGAGCAAGCGTAGTTCACTA
3006 SEQ ID NO: 4280 -8.8 -35.6 86.3 -23.6 -3.2 -12.1
TTTCACACAGATGATTGATT
3118 SEQ ID NO:4281 -8.8 -29.4 75.1 -20.6 0 -5.1
ATCAGTAGCTGAGCTTTCCT
3672 SEQ ID NO:4282 -8.8 -36.4 86.4 -24.8 -1.5 -13.8
GCTTCTATTTTGTTTAAAAT
3804 SEQ ID NO:4283 -8.8 -23.1 67.8 -14.3 0 -4.3
CAGCTTCTATTTTGTTTAAA
3806 SEQ ID NO: 4284 -8.8 -25.3 71.8 -16.5 0 -4
TACAGCTTCTATTTTGTTTA
3808 SEQ ID NO: 4285 -8.8 -27 74.5 -18.2 0 -4
ATAGAAATCAAATTTCTTGT
3894 SEQ ID NO: 4286 -8.8 -24.6 68.6 -14.5 -1.2 -7.8
CAAAGAAAACAAAAACATGT
4691 SEQ ID NO:4287 -8.8 -24.6 70.1 -15.8 0 -5.2
GAGGGAAATATATTTTTTTT
10 SEQ ID NO:4288 -87 -23.4 67.7 -11.9 -0.7 -13.8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CTTTACCTGGGGACCCAGAA
732 SEQ ID NO:4289 -8.7 -39.4 91.6 -23.3 -6.2 -23
CTGGGGCTTGACAAAACCAG
919 SEQ ID NO:4290 -8.7 -37.6 89.1 -25.7 -3.2 -12.3
TTCCAGGTTCATTCCAGCCT
1580 SEQ ID NO:4291 -8.7 -38.3 88.9 -26.6 -3 -8.4
CTGAATTCCTTTTATTTTTT
1616 SEQ ID NO: 4292 -8.7 -22.8 66.8 -14.1 0 -5.2
TTGATTGTCTATATGATCTC
1962 SEQ ID NO: 4293 -8.7 -29.4 74.9 -19.1 -1.6 -8.2
AAATAAAACAACAAAACCTC
2545 SEQ ID NO: 4294 -8.7 -25 71.1 -16.3 0 0
CATGCATAAATCAAAAATGC
2891 SEQ ID NO:4295 -87 -26.7 72.5 -17.1 -0.7 -7
TAAAGGAAATTGTTAAAACC
2982 SEQ ID NO:4296 -8.7 -25.3 72 -16.6 0 -1
GGAATGTGAAAATGGGTGTC
3615 SEQ ID NO:4297 -8.7 -33.8 82.4 -25.1 0 -1.7
TAGAAATCAAATTTCTTGTG
3893 SEQ ID NO:4298 -8.7 -25.6 70.7 -15.8 -1 -7.6
TATTACACAAATAATTTGGC
3922 SEQ ID NO:4299 -8.7 -25.9 72.5 -167 0 -7.6
AACAATCTCATCTAAAAATC
4121 SEQ ID NO: 4300 -87 -26.1 70.8 -17.4 0 0
ATTCAAAGAAAACAAAAACA
4694 SEQ ID NO:4301 -87 -23.6 67.9 -14.9 0 -1.9
AAATAACAAATAACATTCAA
4719 SEQ ID NO:4302 -87 -22.1 65.2 -13.4 0 0
TGTTTTTTTATTAAATAATT
4754 SEQ ID NO:4303 -87 -17.3 567 -8.6 0 -3.7
AGTTCAGAGTCCCCAGAGAA
1260 SEQ ID NO: 4304 -8.6 -38.7 88.8 -30.1 0 -3.1
TTGGTGATGATTTCAGCTAA
2389 SEQ ID NO:4305 -8.6 -31.7 79.7 -20.7 -2.4 -9.7
GAAAGTTCATCACACAGACT
2788 SEQ ID NO: 4306 -8.6 -32.8 807 -24.2 0 -5.6
TTTTACAAACATGGATGTCT
3716 SEQ ID NO: 4307 -8.6 -29.1 76.3 -19.5 0 -9.9
CTACAGCTTCTATTTTGTTT
3809 SEQ ID NO:4308 -8.6 -27.8 75.6 -19.2 0 -4
CTGCACTATTTACATATTGC
4596 SEQ ID NO:4309 -8.6 -29.6 77.8 -20.1 -0.7 -6.2
AACTCTGGCAGAGGAGCCGC
102 SEQ ID NO:4310 -8.5 -43.4 94.8 -29.5 -4.5 -18.8
CAGGAGTTAATGATTCTTTG
141 SEQ ID NO:4311 -8.5 -29.2 76.3 -17.9 -2.1 -13.5
AAAACCAGATCTCCATTATC
907 SEQ ID NO: 4312 -8.5 -30.9 77.9 -22.4 0 -6.5
TCTCTTGCTTAATTACCCCA
999 SEQ ID NO: 4313 -8.5 -34 84.3 -25.5 0 -2.3
TTCTCTTGCTTAATTACCCC
1000 SEQ ID NO:4314 -8.5 -32.8 82.7 -24.3 0 -2.3 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TGTAACTCAGAGGAAACATA
2074 SEQ ID NO:4315 -8.5 -32 807 -22.8 0 -8.8
AGAAGCACCAACCCATTTTC
3134 SEQ ID NO:4316 -8.5 -34.5 84.4 -26 0 -27
AATAAATTGCATGTAAAGCT
4060 SEQ ID NO:4317 -8.5 -26.7 73.3 -16.6 -0.2 -11.4
CTTAGCTTGTGAACGCAGAA
37 SEQ ID NO: 4318 -8.4 -33.9 84 -24.2 -1.2 -9
CCCAGGTCATTTCCCATCAC
421 SEQ ID NO: 4319 -8.4 -38.5 88.3 -29.4 -0.4 -3.4
AGGACTCTCATTCGTCTCTT
749 SEQ ID NO:4320 -8.4 -35.9 84.3 -25.1 -2.4 -6.9
AAGGACTCTCATTCGTCTCT
750 SEQ ID NO:4321 -8.4 -35.9 84.3 -25.1 -2.4 -6.9
CAAGGACTCTCATTCGTCTC
751 SEQ ID NO:4322 -8.4 -35.9 84.1 -25.1 -2.4 -6.9
ATCTCCAAGGACTCTCATTC
756 SEQ ID NO:4323 -8.4 -35.7 83.5 -27.3 0 -6.8
CCCTCCTAACATGTTGAGCG
1817 SEQ ID NO: 4324 -8.4 -37.6 88.2 -27.6 0 -11.3
TCTTCATAAGATACCTGAAG
2098 SEQ ID NO: 4325 -8.4 -31.1 78.7 -21.1 -1.5 -6.6
AAAGCTATTAATTCGACTTT
2496 SEQ ID NO:4326 -8.4 -26.1 72.2 -17.7 0 -5
AAAAGCTATTAATTCGACTT
2497 SEQ ID NO: 4327 -8.4 -26.1 72.2 -17.7 0 -5
AAGCTACGAACTAGCTGCCC
3076 SEQ ID NO:4328 -8.4 -38.9 91.1 -26.5 -4 -11.4
TGGTTATCTGGAATCACAAC
3499 SEQ ID NO:4329 -8.4 -33 81.4 -23.4 -1.1 -7.3
GATGTGTAGTTTTACAAACA
3725 SEQ ID NO:4330 -8.4 -28 75.6 -18 -1.6 -9
TTTTCAGTTTTCTATTACAC
3934 SEQ ID NO:4331 -8.4 -25.7 71.9 -17.3 0 -0.7
TTTAATATTAGATTTTCAGT
3946 SEQ ID NO: 4332 -8.4 -227 66.2 -13.8 0 -7.8
CAATATGATATATATCTGAA
4006 SEQ ID NO:4333 -8.4 -25.1 68.8 -15.1 0 -11.4
CCAATATGATATATATCTGA
4007 SEQ ID NO: 4334 -8.4 -27.5 72.5 -17.5 0 -11.4
ACCAATATGATATATATCTG
4008 SEQ ID NO:4335 -8.4 -27.3 72.6 -17.3 0 -11.4
TTTTTTATTAAATAATTTCT
4751 SEQ ID NO: 4336 -8.4 -17.5 56.8 -9.1 0 -3.7
TAAAAGCTATTAATTCGACT
2498 SEQ ID NO:4337 -8.3 -26.5 73 -18.2 0 -5.3
AAGACAAATAGTTTACCAGC
3441 SEQ ID NO:4338 -8.3 -30.3 79.3 -21.3 -0.5 -3.1
TCACAACTTTTAAGAAGTTA
3486 SEQ ID NO:4339 -8.3 -26.7 73.8 -16.5 -1.7 -11.5
CATGTACAAACTATAAATCA
4202 SEQ ID NO:4340 -8.3 -26.2 72.2 -17.9 0 -7.2 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
TATTAGATGGTGCCTTTAAG
4487 SEQ ID NO:4341 -8.3 -307 79.8 -19.7 0 -13.6
GTGAGTTTTTGGAAACTCCT
593 SEQ ID NO:4342 -8.2 -32.2 81.3 -18.1 -4.2 -20
TTTTTTCTTCGAATTTTATC
1528 SEQ ID NO:4343 -8.2 -22.5 65.5 -14.3 0 -5.9
TCACACAGACTTTGGGCACT
2779 SEQ ID NO:4344 -8.2 -38.1 89.1 -29.9 0 -5.1
TTCCCTTTTCTGATATACAC
2934 SEQ ID NO:4345 -8.2 -30.9 78.9 -22.7 0 -1.6
TCAAAGAAAACAAAAACATG
4692 SEQ ID NO:4346 -8.2 -24.8 70 -16.6 0 -2.2
TTGTTTTTTTATTAAATAAT
4755 SEQ ID NO: 4347 -8.2 -17.3 56.7 -8.6 -0.1 -3.7
TTAGCTTGTGAACGCAGAAG
36 SEQ ID NO:4348 -8.1 -33.9 84 -24.3 -1.4 -8.9
GAGTGAGTTTTTGGAAACTC
595 SEQ ID NO:4349 -8.1 -31.3 79.5 -18.1 -3.4 -18.4
GAAGAAAACTCCAAATCCTG
715 SEQ ID NO:4350 -8.1 -31.3 79.2 -23.2 0 -0.5
ATTAAAAATAGGCTTCTGAT
1169 SEQ ID NO: 4351 -8.1 -27.1 73.4 -19 0 -5
GGCATGCTGGGCAGTTTTTT
1542 SEQ ID NO: 4352 -8.1 -36.4 87.7 -257 -07 -13.3
GTTTTGTCAGTTGATAAAAC
2274 SEQ ID NO: 4353 -8.1 -26.6 73.2 -12.4 -5.9 -19.6
ACTAATAAATTTCACCATCT
2681 SEQ ID NO: 4354 -8.1 -27.2 73.3 -19.1 0 -0.9
TATAAAGGAAATTGTTAAAA
2984 SEQ ID NO:4355 -8.1 -22.2 66.1 -14.1 0 -0.4
TATAACATGTCATGATAAAA
4139 SEQ ID NO:4356 -8.1 -25.4 70 -15.3 0 -12.2
ATATAACATGTCATGATAAA
4140 SEQ ID NO:4357 -8.1 -25.6 69.9 -15.5 0 -12.2
CAATGCTTTCTTCCAAAAGC
492 SEQ ID NO: 4358 -8 -30.8 79.3 -207 -2.1 -6.4
GGGGACCCAGAAGAAAACTC
724 SEQ ID NO:4359 -8 -38.4 89 -26.9 -2.3 -15.1
CGTCTCTTTACCTGGGGACC
737 SEQ ID NO:4360 -8 -40.1 91.5 -25.3 -6.8 -16
CAGAGTCCCCAGAGAAGTCA
1256 SEQ ID NO:4361 -8 -39.9 89.9 -31.9 0 -3.8
GACACCAGGGTAGGGGTGAG
1714 SEQ ID NO:4362 -8 -43.2 95.2 -30.2 -5 -13
AAAACACAGCAGGTTTGCAC
1982 SEQ ID NO:4363 -8 -34 847 -237 -1.7 -12.5
AATAGCTCTTGGCTCTTCAG
2170 SEQ ID NO: 4364 -8 -35.1 84.7 -25.5 -1.5 -8
CTTCCCTTTTCTGATATACA
2935 SEQ ID NO:4365 -8 -30.8 78.8 -22.8 0 -1.4
AAAATGAGCAAGCGTAGTTC
3010 SEQ ID NO:4366 -8 -31.7 80.6 -22.5 -1.1 -7.9 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TGAAAGACAAATAGTTTACC
3444 SEQ ID NO:4367 -8 -28.1 75.7 -19.4 -0.5 -3.1
TCTATTTTGTTTAAAATAAT
3801 SEQ ID NO:4368 -8 -20 61.6 -11.3 -0.1 -8.9
TTCTATTTTGTTTAAAATAA
3802 SEQ ID NO:4369 -8 -19.8 61.6 -11.1 -0.1 -8.9
AAAAATCATTCTTATTTTAC
4108 SEQ ID NO:4370 -8 -21.1 63.5 -13.1 0 -0.7
CTATAAATCATATAGGTTAT
4192 SEQ ID NO:4371 -8 -24.8 69.9 -15 -1.8 -6.2
GTACAAACTATAAATCATAT
4199 SEQ ID NO: 4372 -8 -24.4 69.3 -16.4 0 -5
TGTACAAACTATAAATCATA
4200 SEQ ID NO: 4373 -8 -25.4 71.1 -17.4 0 -6.4
ATGTACAAACTATAAATCAT
4201 SEQ ID NO: 4374 -8 -25.2 70.2 -17.2 0 -7.2
TGTGTAACTCAGAGGAAACA
2076 SEQ ID NO:4375 -7.9 -33.9 83.5 -25 0 -10
ATGCATAAATCAAAAATGCT
2890 SEQ ID NO:4376 -7.9 -26.7 72.3 -16.9 -1.9 -7
TAACTAAATATAAAAATATA
3982 SEQ ID NO:4377 -7.9 -187 59.7 -10.8 0 -1
ATGCAAACTATTGCCAAACT
4566 SEQ ID NO:4378 -7.9 -31 80 -21.2 -1.9 -6.6
AAAATAACAAATAACATTCA
4720 SEQ ID NO:4379 -7.9 -22.1 65.2 -14.2 0 0
GGGAAATATATTTTTTTTTT
8 SEQ ID NO:4380 -7.8 -20.7 63.4 -12.1 0 -9.2
GATCTCCAAGGACTCTCATT
757 SEQ ID NO:4381 -7.8 -35.7 83.5 -27.4 -0.1 -7.1
AGGGAAGTTCAGAGTCCCCA
1265 SEQ ID NO:4382 -7.8 -40.8 92 -29.1 -3.9 -14.8
CGGCATGCTGGGCAGTTTTT
1543 SEQ ID NO:4383 -7.8 -37.9 89.6 -25.4 -1.6 -17.6
TGCACACAGAAAGGGCTACT
3827 SEQ ID NO: 4384 -7.8 -38.2 90 -30.4 0 -5.2
ATACTTTACATACTTTAGTG
4434 SEQ ID NO:4385 -7.8 -26.2 73.4 -18.4 0 -5
GCAGGAGGGAAATATATTTT
14 SEQ ID NO:4386 -7.7 -30.7 78.9 -22.5 0 -7.8
CCCAGAAGAAAACTCCAAAT
719 SEQ ID NO:4387 -77 -32.2 81 -24.5 0 -0.3
AAAAACTGTTCGACCAGGGA
1280 SEQ ID NO:4388 -7.7 -34.9 85.2 -25.1 -1.1 -12.2
CATCACACAGACTTTGGGCA
2781 SEQ ID NO:4389 -7.7 -37 87 -29.3 0 -5.6
TATAAAAGGCACAACTTCCC
2949 SEQ ID NO:4390 -7.7 -32.6 82.8 -24.9 0 -2.4
TATGTAAGAAAAAATGAGCA
3020 SEQ ID NO:4391 -7.7 -27 73.4 -19.3 0 -2.7
AGCTACGAACTAGCTGCCCA
3075 SEQ ID NO:4392 -7.7 -40.1 92.6 -28.7 -3.7 -10.2 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
ACTACAGCTTCTATTTTGTT
3810 SEQ ID NO:4393 -77 -29.1 77.4 -21.4 0 -3.7
AGTTTTCTATTACACAAATA
3929 SEQ ID NO:4394 -7.7 -24.8 70.4 -17.1 0 -0.7
AACTAAATATAAAAATATAT
3981 SEQ ID NO: 4395 -7.7 -18.5 58.9 -10.8 0 -1.4
AGAAAACTCCAAATCCTGCA
713 SEQ ID NO: 4396 -7.6 -33.5 82.6 -25.9 0 -2.7
AGTCCCCAGAGAAGTCAAGT
1253 SEQ ID NO:4397 -7.6 -38.5 89 -30.9 0 -3.8
AAGTTCAGAGTCCCCAGAGA
1261 SEQ ID NO:4398 -7.6 -38.7 88.8 -31.1 0 -3.1
GTGTAACTCAGAGGAAACAT
2075 SEQ ID NO:4399 -7.6 -32.9 81.6 -23.7 0 -11.4
TATACAATAAAAGCTATTAA
2505 SEQ ID NO:4400 -7.6 -23 67.9 -15.4 0 -5.9
AACATGGATGTCTGACATAC
3709 SEQ ID NO:4401 -7.6 -33.3 81 -23.4 -0.1 -12.8
AAACATGGATGTCTGACATA
3710 SEQ ID NO: 4402 -7.6 -32 79.4 -22.1 -0.1 -12.8
AACAGTGAAGAAATTCACAG
3777 SEQ ID NO: 4403 -7.6 -30 77.4 -15.4 -7 -14
GTTTTCTATTACACAAATAA
3928 SEQ ID NO:4404 -7.6 -23.6 68.7 -16 0 -0.7
AATATAAAAATATATTAGAA
3976 SEQ ID NO: 4405 -7.6 -18.7 58.9 -10.3 -0.6 -5.1
AACTATAACTAAATATAAAA
3987 SEQ ID NO: 4406 -7.6 -20.6 63.6 -13 0 -1.2
AAACAATCTCATCTAAAAAT
4122 SEQ ID NO: 4407 -7.6 -24.6 68.6 -17 0 0
AAAACAATCTCATCTAAAAA
4123 SEQ ID NO:4408 -7.6 -24.4 68.6 -16.8 0 0
AAAGAAAACAAAAACATGTC
4690 SEQ ID NO: 4409 -7.6 -24.9 70.2 -17.3 0 -6.2
ATTCATGTCATAGTGGTACA
1124 SEQ ID NO: 4410 -7.5 -32.2 80.2 -23.8 -0.7 -6.2
TGCTGGGCAGTTTTTTCTTC
1538 SEQ ID NO:4411 -7.5 -34.3 84.6 -24.8 -0.4 -12.1
TTTCAGTTTTCTATTACACA
3933 SEQ ID NO:4412 -7.5 -26.9 73.6 -19.4 0 -0.5
ACTATAAATCATATAGGTTA
4193 SEQ ID NO:4413 -7.5 -25.9 71.9 -16.1 -2.3 -7
TACAAACTATAAATCATATA
4198 SEQ ID NO:4414 -7.5 -23.5 67.8 -16 0 -1
GCATGTACAAACTATAAATC
4203 SEQ ID NO:4415 -7.5 -27.5 74.4 -20 0 -7.2
GTAACCCGCTATTAGATGGT
4496 SEQ ID NO:4416 -7.5 -34.8 85.3 -26.1 -0.4 -10.2
ATATTTTTTTTTTCTAAAAA
1 SEQ ID NO: 4417 -7.4 -16.9 55.6 -9.5 0 -0.5
CTTGTGAACGCAGAAGGAGC
32 SEQ ID NO:4418 -7.4 -37.4 87.6 -27.5 -2.5 -9.4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TCTCCAAGGACTCTCATTCG
755 SEQ ID NO:4419 -7.4 -37 85.7 -29.6 0 -6.8
AAAAAACTGTTCGACCAGGG
1281 SEQ ID NO: 4420 -7.4 -33.4 83.6 -23.6 -1.6 -12.7
GAAAAAACTGTTCGACCAGG
1282 SEQ ID NO: 4421 -7.4 -32.5 81.7 -22.7 -1.6 -12.7
TGCCGCCCTCCTAACATGTT
1822 SEQ ID NO: 4422 -7.4 -39.8 91.8 -32.4 0 -6.8
ATACAATAAAAGCTATTAAT
2504 SEQ ID NO: 4423 -7.4 -22.8 67 -15.4 0 -5.9
AGAAAGTTCATCACACAGAC
2789 SEQ ID NO: 4424 -7.4 -32.8 80.7 -25.4 0 -5.6
ACTATAACTAAATATAAAAA
3986 SEQ ID NO:4425 -7.4 -20.6 63.6 -13.2 0 -1.2
AACTATAAATCATATAGGTT
4194 SEQ ID NO: 4426 -7.4 -25.5 71.1 -15.8 -2.3 -7
CTATGCAAACTATTGCCAAA
4568 SEQ ID NO:4427 -7.4 -30.1 79 -20.8 -1.9 -6.6
TTTACCTGGGGACCCAGAAG
731 SEQ ID NO: 4428 -7.3 -39.4 91.6 -24.8 -5.9 -22.7
GCAGTCACTTTTGATGAAAC
2451 SEQ ID NO:4429 -7.3 -31.3 79.2 -22.9 -1 -5
GATATATTAACTAATAAATT
2690 SEQ ID NO: 4430 -7.3 -20 61.8 -12.7 0 -4.8
ACTAAATATAAAGGAAATTG
2990 SEQ ID NO: 4431 -7.3 -23.6 68.4 -16.3 0 -1.5
CACTAAATATAAAGGAAATT
2991 SEQ ID NO: 4432 -7.3 -23.6 68.4 -16.3 0 -1.5
ATTTTCACACAGATGATTGA
3120 SEQ ID NO:4433 -7.3 -29.4 75.1 -22.1 0 -3.4
TGTGTAGTTTTACAAACATG
3723 SEQ ID NO: 4434 -7.3 -27.7 75.5 -19 -1.3 -9
GTGCACACAGAAAGGGCTAC
3828 SEQ ID NO: 4435 -7.3 -38.3 89.6 -29.4 0 -11.3
CAGTTTTCTATTACACAAAT
3930 SEQ ID NO: 4436 -7.3 -25.6 71.6 -18.3 0 -07
AATATTAGATTTTCAGTTTT
3943 SEQ ID NO:4437 -7.3 -22.3 65.3 -15 0 -2.1
TAATATTAGATTTTCAGTTT
3944 SEQ ID NO: 448 -7.3 -22.7 66.2 -15.4 0 -4.4
TTAATATTAGATTTTCAGTT
3945 SEQ ID NO:4439 -7.3 -22.7 66.2 -15.4 0 -5.6
CCAGAAGAAAACTCCAAATC
718 SEQ ID NO:4440 -7.2 -31.3 79.2 -24.1 0 -0.3
ACCTGGGGACCCAGAAGAAA
728 SEQ ID NO:4441 -7.2 -40.5 92.4 -25.9 -6.2 -23
TTACCTGGGGACCCAGAAGA
730 SEQ ID NO: 4442 -7.2 -40.9 93 -26.3 -6.2 -23
GGTCTGATCTCCAAGGACTC
762 SEQ ID NO:4443 -7.2 -39.2 88.1 -29.2 -2.8 -10.4
AATAGCCATTAGAAAAAACT
1293 SEQ ID NO: 4444 -7.2 -26.7 73.6 -19.5 0 -1.5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecu Site oligo binding formation Duplex structure oligo oligo
GCCGCCCTCCTAACATGTTG
1821 SEQ ID NO:4445 -7.2 -39.8 91.4 -32 0 -8.6
TCACTAAATATAAAGGAAAT
2992 SEQ ID NO: 4446 -7.2 -25.1 70.3 -17.9 0 -2.4
AGTTCACTAAATATAAAGGA
2995 SEQ ID NO:4447 -7.2 -27.4 74.3 -20.2 0 -2.1
ATGAGCAAGCGTAGTTCACT
3007 SEQ ID NO: 4448 -7.2 -35.4 85.3 -24.7 -3.5 -13
AAAAAATGAGCAAGCGTAGT
3012 SEQ ID NO:4449 -7.2 -30.2 79 -23 0 -4.8
TTTTCACACAGATGATTGAT
3119 SEQ ID NO: 4450 -7.2 -29.4 75.1 -22.2 0 -5.1
ATATTAGATTTTCAGTTTTC
3942 SEQ ID NO: 4451 -7.2 -23.8 67.7 -16.6 0 -2.1
ACTAAATATAAAAATATATT
3980 SEQ ID NO: 4452 -7.2 -18.5 58.9 -10.8 -0.2 -3.2
AGCTTTAAGTCTGTTTCCCC
475 SEQ ID NO: 4453 -7.1 -34.7 85.5 -27.6 0 -57
AAGAAAACTCCAAATCCTGC
714 SEQ ID NO: 4454 -7.1 -32.3 81.1 -25.2 0 -0.5
TCATGTCATAGTGGTACATC
1122 SEQ ID NO: 4455 -7.1 -337 81.7 -25.9 -0.5 -6.5
ACCTGAAGCCTGTGTAACTC
2086 SEQ ID NO:4456 -7.1 -37.3 88.2 -30.2 0 -1.5
AGATACCTGAAGCCTGTGTA
2090 SEQ ID NO:4457 -7.1 -36.6 87.2 -29.5 0 -3.9
GTTCACTAAATATAAAGGAA
2994 SEQ ID NO:4458 -7.1 -26.2 72.6 -19.1 0 -3
GAAAGACAAATAGTTTACCA
3443 SEQ ID NO:4459 -7.1 -28.1 75.7 -20.3 -0.5 -3.1
CACAACTTTTAAGAAGTTAT
3485 SEQ ID NO:4460 -7.1 -25.4 71.8 -16.4 -1.7 -11.5
TGAAACTATAACTAAATATA
3990 SEQ ID NO: 4461 -7.1 -23.3 67.9 -16.2 0 -1
GGTTTTCTTCTCTACCAGGA
156 SEQ ID NO: 4462 -7 -35.2 85.2 -26.6 -1.5 -7
CAGCCAGTGAGGGTGAAGAC
261 SEQ ID NO:4463 -7 -407 91.4 -30.3 -3.4 -8.6
ATGCTGGGCAGTTTTTTCTT
1539 SEQ ID NO:4464 -7 -33 82.9 -23.6 -0.4 -12.9
TAATTTCATCAAATAGCTCT
2181 SEQ ID NO:4465 -7 -27.4 73.2 -20.4 0 -3.8
ATAAAGGAAATTGTTAAAAC
2983 SEQ ID NO: 4466 -7 -23.1 67.8 -16.1 0 -0.4
CTATTTTGTTTAAAATAATT
3800 SEQ ID NO: 4467 -7 -18.5 59.6 -10.8 -0.1 -8.9
TTTTCTATTACACAAATAAT
3927 SEQ ID NO: 4468 -7 -22.5 66.1 -15.5 0 -0.7
CATATTTTTAATATTAGATT
3952 SEQ ID NO: 4469 -7 -19.5 60.6 -11.8 0 -87
CTATAACTAAATATAAAAAT
3985 SEQ ID NO:4470 -7 -19.5 61.5 -12.5 0 -1.2 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
ACAAACTATAAATCATATAG
4197 SEQ ID NO:4471 -7 -24.3 69.2 -16.6 -0.5 -4.1
TTTTTTTATTAAATAATTTC
4752 SEQ ID NO:4472 -7 -16.3 54.9 -9.3 0 -3.5
ATTGTTTTTTTATTAAATAA
4756 SEQ ID NO:4473 -7 -17.3 56.7 -10.3 0 -2.8
ATCAACTCTGGCAGAGGAGC
105 SEQ ID NO: 4474 -6.9 -39.9 89.8 -27.9 -2.4 -18.3
AAGCTTTAAGTCTGTTTCCC
476 SEQ ID NO: 4475 -6.9 -32.3 82.2 -25.4 0 -5.7
AAAACTGTTCGACCAGGGAA
1279 SEQ ID NO: 4476 -6.9 -34.9 85.2 -25.6 -1.6 -12.7
CCTCCAACAGTGACACCAGG
1725 SEQ ID NO: 4477 -6.9 -40.4 91.6 -33.5 0 -5.7
TACCTGAAGCCTGTGTAACT
2087 SEQ ID NO: 4478 -6.9 -36.2 87.7 -29.3 0 -2.1
TACAATAAAAGCTATTAATT
2503 SEQ ID NO: 4479 -6.9 -22.6 67 -15.7 0 -5.6
TTTATACAATAAAAGCTATT
2507 SEQ ID NO:4480 -6.9 -22.6 67 -157 0 -5
GATAGTTTATACAATAAAAG
2512 SEQ ID NO:4481 -6.9 -22.9 67.6 -16 0 -2
CAACTCTGGCAGAGGAGCCG
103 SEQ ID NO: 4482 -6.8 -42.1 93.2 -29.9 -4.5 -18.8
AGTGAATATCAACTCTGGCA
112 SEQ ID NO: 4483 -6.8 -34.2 82.9 -27.4 0 -4.6
TTCATGTCATAGTGGTACAT
1123 SEQ ID NO: 4484 -6.8 -32.2 80.2 -24.5 -0.7 -6.5
ATGTAAGAAAAAATGAGCAA
3019 SEQ ID NO: 4485 -6.8 -26.6 72.6 -19.8 0 -2.8
CTATTACACAAATAATTTGG
3923 SEQ ID NO:4486 -6.8 -24.6 70.5 -17.3 0 -7.6
AAACTATAACTAAATATAAA
3988 SEQ ID NO: 4487 -6.8 -20.6 63.6 -13.8 0 -1.2
AGCACCACCTTCCTGTCTCC
4461 SEQ ID NO:4488 -6.8 -41.9 93.1 -35.1 0 -27
GAAAGCAAACAGTTTTCATC
790 SEQ ID NO: 4489 -67 -28.8 75.9 -17.2 -4.9 -11.4
GGGGCTTGACAAAACCAGAT
917 SEQ ID NO:4490 -6.7 -36.9 87.5 -28.7 -0.8 -10.8
GTCCCCAGAGAAGTCAAGTT
1252 SEQ ID NO: 4491 -67 -37.3 87.4 -30.6 0 -3.8
GAATTCCTTTTATTTTTTTC
1614 SEQ ID NO: 4492 -6.7 -21.9 65.2 -15.2 0 -5.2
TGAATTCCTTTTATTTTTTT
1615 SEQ ID NO: 4493 -67 -21.6 64.3 -14.9 0 -5.2
AATACTCATGGTCTTATCCA
2354 SEQ ID NO: 4494 -67 -32.3 80.3 -24.2 -1.3 -5.1
ACTTCCCTTTTCTGATATAC
2936 SEQ ID NO:4495 -6.7 -30.9 79 -24.2 0 -1.4
TTCACTAAATATAAAGGAAA
2993 SEQ ID NO:4496 -6.7 -24.9 70.4 -17.7 -0.1 -4.2 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
CATTTTCACACAGATGATTG
3121 SEQ ID NO:4497 -67 -29.1 75.2 -21.4 -0.9 -4.2
TACTACAGCTTCTATTTTGT
3811 SEQ ID NO:4498 -6.7 -29.5 78.2 -22.8 0 -4
GCACCACCTTCCTGTCTCCT
4460 SEQ ID NO:4499 -6.7 -41.9 93.1 -35.2 0 -2.7
CAGGGAAGTTCAGAGTCCCC
1266 SEQ ID NO:4500 -6.6 -40.8 91.6 -30.3 -3.9 -14.8
CAGTGACACCAGGGTAGGGG
1718 SEQ ID NO:4501 -6.6 -42.9 95.3 -35 -1.2 -8.9
TAAGATACCTGAAGCCTGTG
2092 SEQ ID NO:4502 -6.6 -35.3 85.6 -28.7 0 -1.3
ATCACTGAAAAGTGATGACG
2653 SEQ ID NO:4503 -6.6 -32 79.3 -18 -7.4 -14.8
GCTACGAACTAGCTGCCCAT
3074 SEQ ID NO: 4504 -6.6 -39.1 907 -29.9 -2.6 -8
GCTTGTGAACGCAGAAGGAG
33 SEQ ID NO:4505 -6.5 -37.4 87.6 -28.2 -2.7 -9.7
CACTGCTGGGGTTTTCTTCT
165 SEQ ID NO:4506 -6.5 -367 87.6 -28.7 -1.4 -5.7
AAAGCTTTAAGTCTGTTTCC
477 SEQ ID NO:4507 -6.5 -29.9 78.7 -22.9 0 -7.5
AAAAGCTTTAAGTCTGTTTC
478 SEQ ID NO:4508 -6.5 -27.5 75.1 -20.4 0 -8.6
TGATCTCCAAGGACTCTCAT
758 SEQ ID NO:4509 -6.5 -36.9 85 -29.6 -0.6 -7.8
CATGTCATAGTGGTACATCT
1121 SEQ ID NO:4510 -6.5 -33.4 81.7 -26.4 -0.2 -6.5
AAAATAAAACAACAAAACCT
2546 SEQ ID NO:4511 -6.5 -23.5 68.8 -17 0 0
GCATAAATCAAAAATGCTAT
2888 SEQ ID NO:4512 -6.5 -25.9 71.4 -17.5 -1.9 -7
AAAGCTACGAACTAGCTGCC
3077 SEQ ID NO: 4513 -6.5 -36.5 88.1 -26 -4 -11.4
TCTCAGCTGTGTTACAGCTG
3517 SEQ ID NO: 4514 -6.5 -37.3 87.8 -22.4 -8.2 -24.4
CTACAAAAAATATATAACAT
4151 SEQ ID NO:4515 -6.5 -21.6 65 -15.1 0 -3.4
CTGTCTCCTGTTTACATACT
4449 SEQ ID NO:4516 -6.5 -33 82.4 -26.5 0 -4.4
GCTATTAGATGGTGCCTTTA
4489 SEQ ID NO:4517 -6.5 -33.2 83.2 -25.3 -0.7 -10.6
ATACTCATGGTCTTATCCAA
2353 SEQ ID NO:4518 -6.4 -32.3 80.3 -23.8 -2.1 -6.8
TGAAACAGAAGTTTTTTGAT
2437 SEQ ID NO:4519 -6.4 -26.3 71.9 -18.8 -1 -7.4
ATGAAACAGAAGTTTTTTGA
2438 SEQ ID NO: 4520 -6.4 -26.3 71.9 -18.8 -1 -7.9
GATGAAACAGAAGTTTTTTG
2439 SEQ ID NO: 4521 -6.4 -26.3 72.2 -18.8 -1 -6.2
AGGAAATTGTTAAAACCAGA
2979 SEQ ID NO: 4522 -6.4 -28.8 767 -22.4 0 -2.5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular 1molecul Site oligo binding formation Duplex structure oligo oligo
GAAACTATAACTAAATATAA
3989 SEQ ID NO: 4523 -6.4 -22.1 66.2 -15.7 0 -1.2
TGCAGTAGCCCTTCCCTTCC
4041 SEQ ID NO: 4524 -6.4 -41.9 94.2 -35 -0.1 -4.1
TGGGGACCCAGAAGAAAACT
725 SEQ ID NO:4525 -6.3 -38.1 89.4 -27.6 -3 -16.5
AAAAATAGGCTTCTGATCCT
1166 SEQ ID NO: 4526 -6.3 -31.6 79.9 -25.3 0 -6.3
GTTTTTTCTTCGAATTTTAT
1529 SEQ ID NO: 4527 -6.3 -22.3 65.5 -16 0 -5.9
AAGATACCTGAAGCCTGTGT
2091 SEQ ID NO: 4528 -6.3 -36.2 86.6 -29!9 0 -2
CTCCCATCACTGAAAAGTGA
2658 SEQ ID NO:4529 -6.3 -35 83.9 -22.8 -5.9 -11.8
GAAATTGTTAAAACCAGACA
2977 SEQ ID NO:4530 -6.3 -27.7 74.9 -21.4 0 -2
GTAAGAAAAAATGAGCAAGC
3017 SEQ ID NO:4531 -6.3 -28.9 76.8 -22.6 0 -2.8
ATAAGATACCTGAAGCCTGT
2093 SEQ ID NO:4532 -6.2 -34.3 83.9 -28.1 0 -1.3
GTCACTTTTGATGAAACAGA
2448 SEQ ID NO:4533 -6.2 -30.3 77.4 -23.2 -0.7 -4.4
CATAAATCAAAAATGCTATC
2887 SEQ ID NO: 4534 -6.2 -24.9 69.6 -18.7 0 -17
AAAGACAAATAGTTTACCAG
3442 SEQ ID NO:4535 -6.2 -27.8 757 -20.9 -0.5 -3.1
TCAGTTTTCTATTACACAAA
3931 SEQ ID NO:4536 -6.2 -26.9 73.6 -20.7 0 -0.7
AAATATAAAAATATATTAGA
3977 SEQ ID NO:4537 -6.2 -187 58.9 -11.5 -0.9 -5.1
CAAACTATAAATCATATAGG
4196 SEQ ID NO:4538 -6.2 -25.4 71.2 -16.9 -2.3 -7
AGGCAGTCACTTTTGATGAA
2453 SEQ ID NO:4539 -6.1 -33.6 82.5 -26.6 -0.7 -6.2
GGAAATTGTTAAAACCAGAC
2978 SEQ ID NO: 4540 -6.1 -28.9 76.9 -22.8 0 -2.5
TTTCTATTACACAAATAATT
3926 SEQ ID NO: 4541 -6.1 -22.5 66.1 -16.4 0 -0.7
TATTAGATTTTCAGTTTTCT
3941 SEQ ID NO:4542 -6.1 -24.8 69.5 -18.7 0 -1.9
AAACTATAAATCATATAGGT
4195 SEQ ID NO: 4543 -6.1 -25.5 71.1 -17.1 -2.3 -7
AAGAAAACAAAAACATGTCC
4689 SEQ ID NO: 4544 -6.1 -27.3 74.1 -21.2 0 -6.3
AACAAATAACATTCAAAAAG
4715 SEQ ID NO:4545 -6.1 -22.7 66.7 -16.6 0 0
TAAAAATAGGCTTCTGATCC
1167 SEQ ID NO: 4546 -6 -30.8 79 -24.8 0 -5
AAAAATAAAACAACAAAACC
2547 SEQ ID NO: 4547 -6 -22.3 67 -16.3 0 0
AACTTCCCTTTTCTGATATA
2937 SEQ ID NO: 4548 -6 -29.6 77.2 -23.6 0 -1.4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
CCATTTTCACACAGATGATT
3122 SEQ ID NO:4549 -6 -30.3 77 -23 -1.2 -4.9
TATTTTGTTTAAAATAATTT
3799 SEQ ID NO:4550 -6 -17.3 56.7 -10.8 0 -8.1
TTTTAATATTAGATTTTCAG
3947 SEQ ID NO: 4551 -6 -21.4 64.2 -14.7 0 -8.7
TAAAAATCATTCTTATTTTA
4109 SEQ ID NO: 4552 -6 -20.2 61.6 -14.2 0 -0.7
GAGGTACTCACACCATGAAC
1093 SEQ ID NO: 4553 -5.9 -36.6 86.1 -28.4 -2.3 -8.5
GAGTCCCCAGAGAAGTCAAG
1254 SEQ ID NO: 4554 -5.9 -387 88.5 -32.8 0 -3.8
GGGAAGTTCAGAGTCCCCAG
1264 SEQ ID NO:4555 -5.9 -40.8 91.6 -31.2 -3.7 -14.4
CCTGAAGCCTGTGTAACTCA
2085 SEQ ID NO:4556 -5.9 -37.2 88.1 -31.3 0 -2.6
CAAATAGCTCTTGGCTCTTC
2172 SEQ ID NO: 4557 -5.9 -33.9 83.2 -26.6 -1.3 -7.8
CTTTTGATGAAACAGAAGTT
2444 SEQ ID NO: 4558 -5.9 -27.5 73.9 -18.3 -3.3 -8.4
AACTAATAAATTTCACCATC
2682 SEQ ID NO: 4559 -5.9 -26 71.7 -20.1 0 -0.9
CAACTTCCCTTTTCTGATAT
2938 SEQ ID NO: 4560 -5.9 -30.4 78.1 -24.5 0 -1.4
ACAAAAAATATATAACATGT
4149 SEQ ID NO: 4561 -5.9 -22.5 66.1 -16.6 0 -5.2
TACAAAAAATATATAACATG
4150 SEQ ID NO: 4562 -5.9 -21.6 65 -15.7 0 -3.4
CAAAAGCTTTAAGTCTGTTT
479 SEQ ID NO:4563 -5.8 -27.2 75 -20.8 0 -8.6
AAATAGCTCTTGGCTCTTCA
2171 SEQ ID NO:4564 -5.8 -33.9 83.3 -26.5 -1.5 -8
ATCAAATAGCTCTTGGCTCT
2174 SEQ ID NO:4565 -5.8 -34.1 83 -267 -1.5 -8
AAATAATTTTCAACAGTGAA
3788 SEQ ID NO: 4566 -5.8 -24.2 68.6 -18.4 0 -4.6
CTAAATATAAAAATATATTA
3979 SEQ ID NO: 4567 -5.8 -17.6 57.7 -10.8 -0.9 -5.1
ATGTCATGATAAAACAATCT
4133 SEQ ID NO:4568 -5.8 -27.3 72.3 -19.5 0 -12.2
CTATTAGATGGTGCCTTTAA
4488 SEQ ID NO: 4569 -5.8 -30.7 79.8 -23.1 0 -11.8
CATTCAAAGAAAACAAAAAC
4695 SEQ ID NO: 4570 -5.8 -23.6 68.4 -17.8 0 -1.9
TACCTGGGGACCCAGAAGAA
729 SEQ ID NO: 4571 -57 -40.9 93 -27.8 -6.2 -23
CTGAAGCCTGTGTAACTCAG
2084 SEQ ID NO:4572 -5.7 -36 86.4 -29.6 -0.4 -8.2
TCAAATAGCTCTTGGCTCTT
2173 SEQ ID NO:4573 -5.7 -33.9 83.3 -26.6 -1.5 -8
AGTCACTTTTGATGAAACAG
2449 SEQ ID NO:4574 -5.7 -30 77.4 -23.2 -1 -5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TGTAAGAAAAAATGAGCAAG
3018 SEQ ID NO:4575 -57 -27.6 747 -21.9 0 -2.8
GGAGAAAGCAAACAGTTTTC
793 SEQ ID NO:4576 -5.6 -31 79.6 -22 -3.4 -8.4
GTGACACCAGGGTAGGGGTG
1716 SEQ ID NO-.4577 -5.6 -43 95.3 -33.5 -3.9 -11.3
ATAGTTTATACAATAAAAGC
2511 SEQ ID NO:4578 -5.6 -23.9 69.4 -18.3 0 -2.2
ATGTGTAGTTTTACAAACAT
3724 SEQ ID NO:4579 -5.6 -26.7 73.5 -19.5 -1.6 -9
ATTGAGTAAACTAAACCTGC
4405 SEQ ID NO:4580 -5.6 -30.3 79.3 -24.7 0 -3.4
GATTGTTTTTTTATTAAATA
4757 SEQ ID NO:4581 -5.6 -18.8 59.8 -13.2 0 -2
TCAACTCTGGCAGAGGAGCC
104 SEQ ID NO:4582 -5.5 -42.1 92.9 -31.3 -2.9 -18.8
TGAGTTTTTGGAAACTCCTT
592 SEQ ID NO: 4583 -5.5 -30.9 79.6 -19.5 -4.2 -20
CAAAACCAGATCTCCATTAT
908 SEQ ID NO: 4584 ■ -5.5 -30.6 77.9 -25.1 0 -6.5
GGAAGTTCAGAGTCCCCAGA
1263 SEQ ID NO:4585 -5.5 -39.9 90.2 -33.7 -0.4 -7.8
GCTGGGCAGTTTTTTCTTCG
1537 SEQ ID NO:4586 -5.5 -34.6 85 -27.7 -0.2 -10.7
GATACCTGAAGCCTGTGTAA
2089 SEQ ID NO:4587 -5.5 -35.4 85.6 -29.9 0 -3.9
GGCAGTCACTTTTGATGAAA
2452 SEQ ID NO:4588 -5.5 -32.4 80.9 -25.8 -1 -6.2
GAAAAAATGAGCAAGCGTAG
3013 SEQ ID NO:4589 -5.5 -30.4 78.9 -24.9 0 -4.8
GAAAGCTACGAACTAGCTGC
3078 SEQ ID NO:4590 -5.5 -35.6 86.3 -26.1 -4 -11.4
TGGGGCTTGACAAAACCAGA
918 SEQ ID NO:4591 -5.4 -37.9 89.4 -30.9 -1.4 -10.8
AGAGGTACTCACACCATGAA
1094 SEQ ID NO:4592 -5.4 -36.5 86.3 -28.8 -2.3 -8.5
AAGGAAATTGTTAAAACCAG
2980 SEQ ID NO: 4593 -5.4 -27.3 74.9 -21.9 0 -2.5
AAAAATGAGCAAGCGTAGTT
3011 SEQ ID NO: 4594 -5.4 -30.2 79 -24.8 0 -4.8
GCTGGTTATCTGGAATCACA
3501 SEQ ID NO:4595 -5.4 -35.4 84.5 -28.5 -1.4 -8.1
GAATGTGAAAATGGGTGTCT
3614 SEQ ID NO:4596 -5.4 -32.6 80.7 -27.2 0 -4.6
TTGAGTAAACTAAACCTGCG
4404 SEQ ID NO:4597 -5.4 -31.6 81.7 -26.2 0 -3.4
AACTTAGCTTGTGAACGCAG
39 SEQ ID NO:4598 -5.3 -337 84.1 -27.2 -1.1 -7.1
GGAAATATATTTTTTTTTTC
7 SEQ ID NO:4599 -5.2 -19.8 61.7 -14.1 0 -8.2
TTTGTTTAAAATAATTTTCA
3796 SEQ ID NO:4600 -5.2 -19.4 60.6 -14.2 0 -5.5 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tmof target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
ATTTTGTTTAAAATAATTTT
3798 SEQ ID NO: 4601 -5.2 -16.9 55.7 -117 0 -5.5
ACAGGACCAGTCTCCATCTC
4279 SEQ ID NO: 4602 -5.2 -40.2 89.7 -32.6 -2.4 -8.8
TATGCAAACTATTGCCAAAC
4567 SEQ ID NO: 4603 -5.2 -30.2 79.1 -23.1 -1.9 -6.6
GTGAATATCAACTCTGGCAG
111 SEQ ID NO: 4604 -5.1 -34.2 82.8 -29.1 0 -6.6
AGAAAGCAAACAGTTTTCAT
791 SEQ ID NO: 4605 -5.1 -28.5 75.7 -18.3 -5.1 -11.4
CAGGTTTGCACTTGATTGTC
1973 SEQ ID NO: 4606 -5.1 -33.4 82.4 -25.7 -2.6 -8.9
TAACTAATAAATTTCACCAT
2683 SEQ ID NO: 4607 -5.1 -24.9 70.3 -19.8 0 -0.9
TTATGTAAGAAAAAATGAGC
3021 SEQ ID NO:4608 -5.1 -25.8 71.7 -20.7 0 -3.4
TTTTGTTTAAAATAATTTTC
3797 SEQ ID NO:4609 -5.1 -18.2 58.8 -13.1 0 -5.5
GCACACAGAAAGGGCTACTA
3826 SEQ ID NO:4610 -5.1 -37.4 89.1 -32.3 0 -5.2
GTTTTCTTCTCTACCAGGAG
155 SEQ ID NO: 4611 -5 . -34 83.4 -27.4 -1.6 -6.1
GCATGCTGGGCAGTTTTTTC
1541 SEQ ID NO: 4612 -5 -35.5 85.8 -28.1 -0.6 -12.9
CTGTGTAACTCAGAGGAAAC
2077 SEQ ID NO: 4613 -5 -33.9 83.3 -27.1 -1.8 -9.8
TTAAGGCAGTCACTTTTGAT
2456 SEQ ID NO:4614 -5 -31.3 80.1 -25.7 -0.3 -6.2
TCTCCCATCACTGAAAAGTG
2659 SEQ ID NO:4615 -5 -35 83.9 -26.2 -3.8 -7.6
CTACTACAGCTTCTATTTTG
3812 SEQ ID NO:4616 -5 -29.4 78.1 -24.4 0 -4
TATAACTAAATATAAAAATA
3984 SEQ ID NO:4617 -5 -18.7 59.7 -13.7 0 -1
CACAGGACCAGTCTCCATCT
4280 SEQ ID NO: 4618 -5 -39.9 89.7 -32.5 -2.4 -8.8
TGTCTCCTGTTTACATACTT
4448 SEQ ID NO:4619 -4.9 -31.8 80.8 -26.9 0 -4.4
ACCAGGAGTTAATGATTCTT
143 SEQ ID NO: 4620 -4.8 -31.7 79.9 -24.5 -2 -12.6
AAGGCAGTCACTTTTGATGA
2454 SEQ ID NO: 4621 -4.8 -33.6 82.5 -27.7 -1 -6.2
AATGAGCAAGCGTAGTTCAC
3008 SEQ ID NO:4622 -4.8 -34.2 83.8 -25.9 -3.5 -13
AGTTTTTTCTTCGAATTTTA
1530 SEQ ID NO:4623 -4.7 -23.3 67.3 -18.6 0 -5.9
CTCAGCTGTGTTACAGCTGG
3516 SEQ ID NO: 4624 -4.7 -38.2 89.3 -23.8 -9.5 -26.7
CGCTATTAGATGGTGCCTTT
4490 SEQ ID NO:4625 -4.7 -34.3 84.4 -26.7 -2.3 -13.8
TATCAACTCTGGCAGAGGAG
106 SEQ ID NO: 4626 -4.6 -37.8 87.5 -27.9 -2.5 -18.8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
GTTTTTTTATTAAATAATTT
4753 SEQ ID NO:4627 -4.6 -16.1 54.9 -11.5 0 -3.7
TACCAGGAGTTAATGATTCT
144 SEQ ID NO: 4628 -4.5 -32.1 80.6 -26 -1.1 -11.1
ACAGTGACACCAGGGTAGGG
1719 SEQ ID NO:4629 -4.5 -41.8 94.1 -36 -1.2 -8.9
AACAGTGACACCAGGGTAGG
1720 SEQ ID NO:4630 -4.5 -39.4 91.3 -34.1 -0.4 -8.9
GAAGCCTGTGTAACTCAGAG
2082 SEQ ID NO:4631 -4.5 -36.3 86.3 -30.5 -0.9 -10.2
TTCTTCTCTACCAGGAGTTA
152 SEQ ID NO:4632 -4.4 -34.4 84.3 -27.8 -2.2 -7.4
GAGAAAGCAAACAGTTTTCA
792 SEQ ID NO:4633 -4.4 -29.8 77.7 -20.3 -5.1 -11.4
AAAATAGGCTTCTGATCCTG
1165 SEQ ID NO:4634 -4.4 -32.8 81.4 -27.2 -1.1 -8.5
CTGGTTATCTGGAATCACAA
3500 SEQ ID NO:4635 -4.4 -32.9 81.3 -27.2 -1.2 -8.1
ATTAGATTTTCAGTTTTCTA
3940 SEQ ID NO:4636 -4.3 -24.8 69.5 -20 -0.1 -3.6
TCCCATCACTGAAAAGTGAT
2657 SEQ ID NO: 4637 -4.2 -34 82.2 -22.8 -7 -14
TAAATATAAAAATATATTAG
3978 SEQ ID NO: 4638 -4.2 -17.6 57.7 -12.4 -0.9 -5.1
TAACCCGCTATTAGATGGTG
4495 SEQ ID NO:4639 -4.2 -34.7 85.2 -27.8 -1.9 -13.3
GGTGATGATTTCAGCTAACA
2387 SEQ ID NO:4640 -4.1 -33 81.3 -26.7 -2.2 -7.4
TGGGTGTCTAGCCATTTTTG
3603 SEQ ID NO: 4641 -4.1 -34.6 85.1 -27.8 -27 -9.9
GTGAAAATGGGTGTCTAGCC
3610 SEQ ID NO: 4642 -4.1 -36.2 86.5 -31 -1 -8.6
AAATTGCATGTAAAGCTGCA
4057 SEQ ID NO: 4643 -4.1 -31 79.9 -24.7 -2.2 -11.4
CTGTATGAATGCATGTACAA
4213 SEQ ID NO: 4644 -4.1 -30.7 78.5 -237 -2.5 -13.5
CAGCAGGTTTGCACTTGATT
1976 SEQ ID NO: 4645 -4 -34.3 84.2 -27.5 -2.8 -12.5
TTAACTAATAAATTTCACCA
2684 SEQ ID NO: 4646 -4 -247 70.4 -20.7 0 -0.9
TCTATTACACAAATAATTTG
3924 SEQ ID NO: 4647 -4 -23.7 68.4 -19.7 0 " -5.2
AGATTGAGTAAACTAAACCT
4407 SEQ ID NO: 4648 -4 -29.3 77.5 -25.3 0 -3.4
TGAAAATAACAAATAACATT
4722 SEQ ID NO:4649 -4 -22.1 65.2 -18.1 0 0
GAAATATATTTTTTTTTTCT
6 SEQ ID NO: 4650 -3.9 -18.6 59 -14.7 0 -7.2
TTTCTTCTCTACCAGGAGTT
153 SEQ ID NO:4651 -3.9 -34 83.6 -27.9 -2.2 -7.4
TTTTCTTCTCTACCAGGAGT
154 SEQ ID NO: 4652 -3.9 -34 83.6 -27.9 -2.2 -7.4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
TCTCTACCAGGAGTTAATGA
148 SEQ ID NO:4653 -3.8 -34.6 83.9 -28.6 -2.2 -6.8
GAGTTTTTGGAAACTCCTTC
591 SEQ ID NO:4654 -3.8 -31.2 79.6 -21.5 -4.2 -20
GAAGTTCAGAGTCCCCAGAG
1262 SEQ ID NO:4655 -3.8 -38.7 88.5 -34.9 0 -6.3
ACCTCCAACAGTGACACCAG
1726 SEQ ID NO:4656 -3.8 -39.3 90.3 -35.5 0 -5.7
TGAAGCCTGTGTAACTCAGA
2083 SEQ ID NO:4657 -3.8 -36.3 86.6 -31 -1.2 -10.5
CAAATGATTGTTTTTTTATT
4762 SEQ ID NO:4658 -3.8 -20.4 62.3 -16.6 0 -6.1
ATGGGTGTCTAGCCATTTTT
3604 SEQ ID NO:4659 -37 -33.6 83.4 -27.2 -2.7 -9.9
AATAATTTTCAACAGTGAAG
3787 SEQ ID NO:4660 -3.7 -25.4 70.8 -20.9 -0.6 -6
ATAACTAAATATAAAAATAT
3983 SEQ ID NO:4661 -37 -18.5 58.9 -14.8 0 -1
CAGGACCAGTCTCCATCTCC
4278 SEQ ID NO: 4662 -3.7 -41.3 90.9 -35.2 -2.4 -8.9
TATATTAACTAATAAATTTC
2688 SEQ ID NO: 4663 -3.6 -19.8 617 -16.2 0 -4
GAAAATGGGTGTCTAGCCAT
3608 SEQ ID NO: 4664 -3.6 -35.1 84.8 -28.8 -2.7 -10.3
TAAATTGCATGTAAAGCTGC
4058 SEQ ID NO:4665 -3.6 -30.2 79 -25 -0.4 -11.4
CTTCTCTACCAGGAGTTAAT
150 SEQ ID NO:4666 -3.5 -33.1 82.4 -27.4 -2.2 -7.2
CCAGGGAAGTTCAGAGTCCC
1267 SEQ ID NO:4667 -3.5 -40.8 91.6 -34.2 -3.1 -13.2
ACTTTTGATGAAACAGAAGT
2445 SEQ ID NO:4668 -3.5 -28.8 75.7 -20.4 -4.9 -11.6
AATGGGTGTCTAGCCATTTT
3605 SEQ ID NO:4669 -3.5 -33.6 83.4 -27.3 -2.8 -11.7
TGTATGAATGCATGTACAAA
4212 SEQ ID NO:4670 -3.4 -29.5 76.8 -23.5 -2.4 -12.7
CCGCTATTAGATGGTGCCTT
4491 SEQ ID NO:4671 -3.4 -36.7 87.6 -30.1 -2.6 -14.4
CAACAGTGACACCAGGGTAG
1721 SEQ ID NO:4672 -3.3 -38.2 89.5 -34.1 -0.4 -8.9
AAATGAGCAAGCGTAGTTCA
3009 SEQ ID NO:4673 -3.2 -32.9 82.2 -26.7 -3 -11.7
AATTGCATGTAAAGCTGCAG
4056 SEQ ID NO:4674 -3.2 -32.2 81.5 -26 -3 -11.4
CTCCAAGGACTCTCATTCGT
754 SEQ ID NO:4675 -3.1 -36.8 85.8 -33.7 0 -6.8
TTAAAAATAGGCTTCTGATC
1168 SEQ ID NO: 4676 -3.1 -28.4 75.4 -25.3 0 -5
GCAGGTTTGCACTTGATTGT
1974 SEQ ID NO: 4677 -3.1 -34.4 84.2 -28.5 -2.8 -12.5
AGCAGGTTTGCACTTGATTG
1975 SEQ ID NO:4678 -3.1 -34.3 84.2 -28.4 -2.8 -12.5 kcal/mol kcal/mol degC kcal/mol kcal/mol kcal/mol Intra Inter
Target total duplex Tm of target molecular molecular
Site oligo binding formation Duplex structure oligo oligo
TCACTTTTGATGAAACAGAA
2447 SEQ ID NO:4679 -3.1 -29 75.5 -25.4 -0.2 -2.9 TGAAAATGGGTGTCTAGCCA
3609 SEQ ID NO:4680 -3.1 -36.1 86.6 -30.3 -2.7 -8.9 TTCAGTTTTCTATTACACAA
3932 SEQ ID NO:4681 -3.1 -26.9 73.6 -23.8 0 -07 TCTTCTCTACCAGGAGTTAA
151 SEQ ID NO:4682 -3 -34.4 84.3 -29.2 -2.2 -7.4 CACTTTTGATGAAACAGAAG
2446 SEQ ID NO:4683 -2.9 -28.7 75.6 -22.6 -3.2 -8.2 CCCGCTATTAGATGGTGCCT
4492 SEQ ID NO:4684 -2.9 -39.1 90.5 -33 -2.6 -14.4 AATATATTTTTTTTTTCTAA
4 SEQ ID NO:4685 -2.8 -17.5 56.8 -14.7 0 -2.8 AAATATATTTTTTTTTTCTA
5 SEQ ID NO:4686 -2.8 -17.5 56.8 -14.7 0 -4.6 CAGTCACTTTTGATGAAACA
2450 SEQ ID NO:4687 -2.8 -30 77.3 -26.1 -1 -5 TAAGGCAGTCACTTTTGATG
2455 SEQ ID NO:4688 -2.8 -32.5 81.6 -28.6 -1 -6.2 ATATTAACTAATAAATTTCA
2687 SEQ ID NO:4689 -2.8 -20.6 62.6 -17.8 0 -4 AAAATGGGTGTCTAGCCATT
3607 SEQ ID NO:4690 -2.8 -33.6 83.4 -27.8 -3 -12.1 CCAGGAGTTAATGATTCTTT
142 SEQ ID NO:4691 -2.7 -30.4 78.1 -24.9 -2.1 -13.5 AAGCCTGTGTAACTCAGAGG
2081 SEQ ID NO:4692 -2.7 -37.2 88.2 -30.5 -4 -11 ATAAATTGCATGTAAAGCTG
4059 SEQ ID NO:4693 -2.7 -27.9 75.2 -23.6 -0.2 -11.1 TGAATATCAACTCTGGCAGA
110 SEQ ID NO: 4694 -2.6 -34.4 82.8 -30.6 0 -10.4 CTGGGGACCCAGAAGAAAAC
726 SEQ ID NO: 4695 -2.6 -38.1 89 -28.4 -5.9 -22.3 CTACCAGGAGTTAATGATTC
145 SEQ ID NO:4696 -2.5 -32.1 80.5 -29.1 0 -7.6 GATTGAGTAAACTAAACCTG
4406 SEQ ID NO:4697 -2.5 -29.3 77.5 -26.8 0 -3 GGGGTTTTCTTCTCTACCAG
158 SEQ ID NO:4698 -2.4 -36.1 86.8 -29.5 -4.2 -8.4 TCCAAGGACTCTCATTCGTC
753 SEQ ID NO:4699 -2.4 -37.1 85.7 -34 -0.4 -6 AGGAGCAAAACACAGCAGGT
1988 SEQ ID NO:4700 -2.4 -37.8 89.3 -34.1 -1.2 -3.1 GAGATTGAGTAAACTAAACC
4408 SEQ ID NO:4701 -2.4 -29.6 77.5 -27.2 0 -3.4 ATATATTTTTTTTTTCTAAA
3 SEQ ID NO: 4702 -2.3 -17.5 56.8 -15.2 0 -1 ACTGCTGGGGTTTTCTTCTC
164 SEQ ID NO:4703 -2.3 -37 87.6 -31.7 -3 -7.2 ATATATTAACTAATAAATTT
2689 SEQ ID NO:4704 -2.3 -18.5 58.9 -16.2 0 -4 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular imolecul Site oligo binding formation Duplex structure oligo oligo
AAATGGGTGTCTAGCCATTT
3606 SEQ ID NO:4705 -2.2 -33.6 83.4 -27.3 -4.1 -14.3
GAAAATAACAAATAACATTC
4721 SEQ ID NO:4706 -2.1 -22.4 66 -20.3 0 0
TTCTCTACCAGGAGTTAATG
149 SEQ ID NO:4707 -2 -33.1 82.4 -28.9 -2.2 -6.8
AATTATGTAAGAAAAAATGA
3023 SEQ ID NO:4708 -2 -22.3 65.2 -20.3 0 -4.2
CCCATCACTGAAAAGTGATG
2656 SEQ ID NO:4709 -1.9 -337 82.1 -22.2 -9.6 -19.2
GGAGATTGAGTAAACTAAAC
4409 SEQ ID NO:4710 -1.9 -29.6 77.5 -277 0 -3.4
AACCCGCTATTAGATGGTGC
4494 SEQ ID NO:4711 -1.8 -36.8 87.6 -31.8 -2.6 -14.4
CTCTACCAGGAGTTAATGAT
147 SEQ ID NO:4712 -1.6 -33.3 82.1 -29.6 -2.1 -6.6
GCCTGTGTAACTCAGAGGAA
2079 SEQ ID NO:4713 -1.6 -37.5 88.1 -30.8 -5.1 -13.2
CTGCAGTAGCCCTTCCCTTC
4042 SEQ ID NO: 4714 -1.6 -40.7 92.5 -38.4 -0.1 -8.9
GCTGGGGTTTTCTTCTCTAC
161 SEQ ID NO:4715 -1.5 -36.2 86.7 -30.5 -4.2 -10.4
TGCTGGGGTTTTCTTCTCTA
162 SEQ ID NO:4716 -1.3 -36.1 86.9 -30.6 -4.2 -10.4
GCTACTACAGCTTCTATTTT
3813 SEQ ID NO: 4717 -1.3 -30.7 80 -27.9 -1.4 -5.4
ATTGCATGTAAAGCTGCAGT
4055 SEQ ID NO:4718 -1.3 -33.5 83.2 -27.7 -4.5 -13.3
TTGCATGTAAAGCTGCAGTA
4054 SEQ ID NO:4719 -1.2 -33.7 84.2 -29.1 -3.2 -14.2
AGCCTGTGTAACTCAGAGGA
2080 SEQ ID NO:4720 -1.1 -38.7 89.6 -32.5 -5.1 -13.2
CACACAGAAAGGGCTACTAC
3825 SEQ ID NO: 4721 -1.1 -36.2 87.4 -35.1 0 -5.2
CTGCTGGGGTTTTCTTCTCT
163 SEQ ID NO: 4722 -1 -36.9 87.5 -31.7 -4.2 -10.4
TGTGAAAATGGGTGTCTAGC
3611 SEQ ID NO: 4723 -0.9 -35 84.9 -33.5 0 -8.6
CCAAGGACTCTCATTCGTCT
752 SEQ ID NO: 4724 -0.8 -36.8 85.8 -33.7 -2.3 -5.9
TATATTTTTTTTTTCTAAAA
2 SEQ ID NO: 4725 -0.7 -17.3 56.6 -16.6 0 0
CCTGGGGACCCAGAAGAAAA
727 SEQ ID NO:4726 -0.7 -39.2 907 -31.1 -6.2 -23
ATTATGTAAGAAAAAATGAG
3022 SEQ ID NO:4727 -07 -23.5 67.6 -22.8 0 -4.2
ATATCAACTCTGGCAGAGGA
107 SEQ ID NO: 4728 -0.5 -36.8 86 -31 -2.4 -18.8
TCTACCAGGAGTTAATGATT
146 SEQ ID NO:4729 -0.5 -32.1 80.6 -31.6 0 -4.4
CCTGTGTAACTCAGAGGAAA
2078 SEQ ID NO:4730 -0.4 -35 85 -29.7 -4.9 -12.8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol
Intra Inter
Target total duplex Tm of target molecular molecul Site oligo binding formation Duplex structure oligo oligo
CTGGGGTTTTCTTCTCTACC
160 SEQ ID NO: 4731 -0.1 -36.1 86.8 -31.8 -4.2 -8.6
AAGCACCAACCCATTTTCAC
3132 SEQ ID NO:4732 -0.1 -34.3 84.5 -34.2 0 -2.7
GAATATCAACTCTGGCAGAG
109 SEQ ID NO:4733 0 -34.4 82.7 -30.3 -1.1 -16.4
AGCACCAACCCATTTTCACA
3131 SEQ ID NO:4734 0 -35.5 86 -35.5 0 -2.7
GGCTACTACAGCTTCTATTT
3814 SEQ ID NO:4735 0 -33.1 83.4 -31.2 -1.9 -6.4
TTCTATTACACAAATAATTT
3925 SEQ ID NO:4736 0.1 -22.5 66.1 -22.6 0 -0.7
ATATTTTTAATATTAGATTT
3951 SEQ ID NO:4737 0.1 -18.3 58 -17.8 0 -8.5
GCATGTAAAGCTGCAGTAGC
4052 SEQ ID NO: 4738 0.3 -36.2 87.1 -33.4 -2.1 -14.2
GAAGCACCAACCCATTTTCA
3133 SEQ ID NO:4739 0.4 -34.5 84.3 -34.9 0 -2.7
TGCATGTAAAGCTGCAGTAG
4053 SEQ ID NO:4740 0.4 -34.9 85.6 -32.1 -2.8 -14.2
TTTTTAATATTAGATTTTCA
3948 SEQ ID NO:4741 0.8 -20.2 61.6 -20.3 0 -8.7
CATCACTGAAAAGTGATGAC
2654 SEQ ID NO: 4742 1.1 -31.7 78.8 -22.4 -10.4 -20.8
TATTTTTAATATTAGATTTT
3950 SEQ ID NO: 4743 1.1 -18.1 57.9 -18.5 0 -8.7
AATATCAACTCTGGCAGAGG
108 SEQ ID NO: 4744 1.3 -35.3 84.5 -31.3 -2.4 -18.8
CATGTAAAGCTGCAGTAGCC
4051 SEQ ID NO: 4745 1.4 -36.1 87.1 -34.2 -1.3 -14.8
TGGGGTTTTCTTCTCTACCA
159 SEQ ID NO: 4746 1.5 -36.1 87 -33.4 -4.2 -8.6
ATTTTTAATATTAGATTTTC
3949 SEQ ID NO: 4747 17 -19.2 60 -20.2 0 -8.7
ATTAACTAATAAATTTCACC
2685 SEQ ID NO: 4748 1.8 -23.7 68.6 -25.5 0 -0.9
GGGCTACTACAGCTTCTATT
3815 SEQ ID NO: 4749 1.9 -35.5 86.6 -35 -2.4 -7.4
CCATCACTGAAAAGTGATGA
2655 SEQ ID NO:4750 2 -32.8 80.4 -24.1 -10.7 -21.4
TATTAACTAATAAATTTCAC
2686 SEQ ID NO:4751 2.1 -21.7 65.3 -23.8 0 -3.2
AGCTGCAGTAGCCCTTCCCT
4044 SEQ ID NO: 4752 2.6 -42.9 95.8 -42.2 -1.3 -14.8
GCTGCAGTAGCCCTTCCCTT
4043 SEQ ID NO: 4753 27 -417 94.3 -41.3 -1.1 -14.4
TGTAAAGCTGCAGTAGCCCT
4049 SEQ ID NO: 4754 3 -38.3 90.9 -38 -1.3 -14.8
ATGTAAAGCTGCAGTAGCCC
4050 SEQ ID NO:4755 3 -37.3 88.9 -37 -1.3 -14.8
AGGGCTACTACAGCTTCTAT
3816 SEQ ID NO: 4756 3.2 -36.7 88.2 -37.2 -2.7 -8.8 kcal/mol kcal/mol deg C kcal/mol kcal/mol kcal/mol Intra Inter
Target total duplex Tm of target molecular molecular Site oligo binding formation Duplex structure oligo oligo
ACACAGAAAGGGCTACTACA 3824 SEQ ID NO: 4757 3.9 -36.2 87.7 -40.1 0 -5.2
GTAAAGCTGCAGTAGCCCTT 4048 SEQ ID NO: 4758 4.2 -37.1 89.3 -38.1 -1.1 -14.6
AAGGGCTACTACAGCTTCTA
3817 SEQ ID NO:4759 4.3 -36.5 88.5 -38.1 -2.7 -8.8 ACAGAAAGGGCTACTACAGC
3822 SEQ ID NO: 4760 4.4 -37.4 89.1 -41.3 -0.2 -5.2 ACCCGCTATTAGATGGTGCC
4493 SEQ ID NO: 4761 4.4 -39.2 90.6 -40.4 -2.6 -14.4
AAAGCTGCAGTAGCCCTTCC
4046 SEQ ID NO: 4762 5 -39.3 91.5 -41 -1.3 -14.8 AAGCTGCAGTAGCCCTTCCC
4045 SEQ ID NO: 4763 5.2 -41.7 94.3 -43.6 -1.3 -14.8
TAAAGCTGCAGTAGCCCTTC
4047 SEQ ID NO: 4764 5.2 -37.3 89.2 -39.2 -1.3 -14.8 AAAGGGCTACTACAGCTTCT
3818 SEQ ID NO:4765 5.8 -36.1 87.9 -39.2 -2.7 -8.8 CACAGAAAGGGCTACTACAG
3823 SEQ ID NO: 4766 6 -36.1 87.3 -42.1 0 -5.2 CAGAAAGGGCTACTACAGCT
3821 SEQ ID NO: 4767 7.1 -37.3 89.1 -42.2 -2.2 -7
GAAAGGGCTACTACAGCTTC
3819 SEQ ID NO: 4768 7.3 -36.4 87.4 -41 -2.7 -8.8 AGAAAGGGCTACTACAGCTT
3820 SEQ ID NO: 4769 9.8 -36.1 87.9 -43.6 -2.3 -7.5
Example 15
Western blot analysis of GR protein levels
[00190] Western blot analysis (immunoblot analysis) is carried out using standard methods. Cells are harvested 16-20 h after oligonucleotide treatment, washed once with PBS, suspended in Laemmli buffer (100 ul/well), boiled for 5 minutes and loaded on a 16% SDS-PAGE gel. Gels are run for 1.5 hours at 150 V, and transferred to membrane for western blotting. Appropriate primary antibody directed to GR is used, with a radiolabelled or fluorescently labeled secondary
10 antibody directed against the primary antibody species. Bands are visualized using a
PHOSPHORIMAGER™ (Molecular Dynamics, Sunnyvale CA).

Claims

WHAT IS CLAIMED IS:
1. An antisense compound targeted to a nucleic acid molecule encoding mammalian glucocorticoid receptor, wherein said antisense compound is a complement to the nucleic acid sequence of Figure 1 or Figure 2.
2. An antisense compound 8 to 30 nucleobases in length targeted to a nucleic acid molecule encoding mammalian glucocorticoid receptor, wherein said antisense compound specifically hybridizes with and inhibits the expression of mammalian glucocorticoid receptor.
3. The antisense compound of claim 2 which is an antisense oligonucleotide.
4. The antisense compound of claim 3 wherein said antisense compound comprise a nucleic acid sequence selected from the group consisiting of SEQ ID NO: 1 -SEQ ID NO:4769, and a fragment of at least eight contiguous nucleotides of SEQ ID NO: 1 -SEQ ID NO:4769.
5. The antisense compound of claim 4 wherein the antisense oligonucleotide comprises at least one modified internucleoside linkage.
6. The antisense compound of claim 5 wherein the modified internucleoside linkage is a phosphorothioate linkage.
7. The antisense compound of claim 3 wherein the antisense oligonucleotide comprises at least one modified sugar moiety.
8. The antisense compound of claim 7 wherein the modified sugar moiety is a 2'-O-methoxyethyl sugar moiety.
9. The antisense compound of claim 3 wherein the antisense oligonucleotide comprises at least one modified nucleobase.
10. The antisense compound of claim 9 wherein the modified nucleobase is a 5-methylcytosine.
1 1. The antisense compound of claim 3 wherein the antisense oligonucleotide is a chimeric oligonucleotide.
12. A composition comprising the antisense compound of claim 2 and a pharmaceutically acceptable carrier or diluent.
13. The composition of claim 12 further comprising a colloidal dispersion system.
14. The composition of claim 13 wherein the antisense compound is an antisense oligonucleotide.
15. A method of inhibiting the expression of mammalian glucocorticoid receptor in cells or tissues comprising contacting said cells or tissues with the antisense compound of claim 2 so that expression of mammalian glucocorticoid receptor is inhibited.
16. A method of treating a human having a disease or condition associated with mammalian glucocorticoid receptor comprising administering to said patient a therapeutically or prophylactically effective amount of the antisense compound of claim 2 so that expression of mammalian glucocorticoid receptor is inhibited.
17. The method of claim 16 wherein the disease or condition is diabetes.
18. The method of claim 16 wherein the disease or condition is obesity.
19. The method of claim 16 wherein the disease or condition is a cardiovascular disorder.
20. The method of claim 16 wherein the disease or condition is a hyperlipidemia.
21. The method of claim 16 wherein the disease is Cushings Syndrom.
EP03729073A 2002-05-20 2003-05-20 Antisense modulation of glucocorticoid receptor expression Withdrawn EP1534728A4 (en)

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EP2363480A3 (en) * 2004-01-20 2015-10-07 Isis Pharmaceuticals, Inc. Modulation of glucocorticoid receptor expression
EP1799859B1 (en) * 2004-09-17 2014-07-02 Isis Pharmaceuticals, Inc. Enhanced antisense oligonucleotides
EP2368988A1 (en) 2005-09-19 2011-09-28 Isis Pharmaceuticals, Inc. Modulation of Glucocorticoid Receptor Expression
JP2009536222A (en) * 2006-05-05 2009-10-08 アイシス ファーマシューティカルズ， インコーポレーテッド Compounds and methods for modulating the expression of PCSK9
JP2009171895A (en) * 2008-01-24 2009-08-06 Hitachi Software Eng Co Ltd Method for analyzing function of intranuclear non-coding rna
EP2429657A2 (en) * 2009-05-15 2012-03-21 F. Hoffmann-La Roche AG Compositions and methods for inhibiting expression of glucocorticoid receptor (gcr) genes
EP2545173A2 (en) 2010-03-12 2013-01-16 Sarepta Therapeutics, Inc. Antisense modulation of nuclear hormone receptors
RU2014119787A (en) 2011-10-25 2015-12-10 Айсис Фармасьютикалс, Инк. GCCR ANTI-SENSE REGULATION
WO2013088853A1 (en) * 2011-12-15 2013-06-20 国立大学法人東京農工大学 Oligonucleotide, glucocorticoid sensitivity enhancer, drug composition, and expression vector
WO2016077704A1 (en) * 2014-11-14 2016-05-19 The Regents Of The University Of California Modulation of agpat5 expression
AU2019269601A1 (en) * 2018-05-17 2020-11-26 Regents Of The University Of Minnesota Drug-resistant immune cells and methods of use thereof

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AU2003233634A1 (en) 2003-12-12
JP2005525829A (en) 2005-09-02

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