Patent Publication Number: US-7897757-B2

Title: RNA interference mediated inhibition of protein tyrosine phosphatase-1B (PTP-1B) gene expression using short interfering nucleic acid (siNA)

Description:
This application is a continuation of U.S. patent application Ser. No. 11/748,029, filed May 14, 2007, which is a continuation of U.S. patent application Ser. No. 11/031,668, filed on Jan. 6, 2005, which is a continuation-in-part of U.S. patent application Ser. No. 10/894,475, filed Jul. 19, 2004, which is a continuation-in-part of International Patent Application No. PCT/US03/04123, filed Feb. 11, 2003, which is a continuation-in-part of U.S. patent application Ser. No. 10/206,705, filed Jul. 26, 2002. This application is also a continuation-in-part of International Patent Application No. PCT/US04/16390, filed May 24, 2004, which is a continuation-in-part of U.S. patent application Ser. No. 10/826,966, filed Apr. 16, 2004, which is a continuation-in-part of U.S. patent application Ser. No. 10/757,803, filed Jan. 14, 2004, which is a continuation-in-part of U.S. patent application Ser. No. 10/720,448, filed Nov. 24, 2003, which is a continuation-in-part of U.S. patent application Ser. No. 10/693,059, filed Oct. 23, 2003, which is a continuation-in-part of U.S. patent application Ser. No. 10/444,853, filed May 23, 2003, which is a continuation-in-part of International Patent Application No. PCT/US03/05346, filed Feb. 20, 2003, and a continuation-in-part of International Patent Application No. PCT/US03/05028, filed Feb. 20, 2003, both of which claim the benefit of U.S. Provisional Application No. 60/358,580, filed Feb. 20, 2002, U.S. Provisional Application No. 60/363,124, filed Mar. 11, 2002, U.S. Provisional Application No. 60/386,782, filed Jun. 6, 2002, U.S. Provisional Application No. 60/406,784, filed Aug. 29, 2002, U.S. Provisional Application No. 60/408,378, filed Sep. 5, 2002, U.S. Provisional Application No. 60/409,293, filed Sep. 9, 2002, and U.S. Provisional Application No. 60/440,129, filed Jan. 15, 2003. The instant application claims the benefit of all the listed applications, which are hereby incorporated by reference herein in their entireties, including the drawings. 
    
    
     SEQUENCE LISTING 
     The sequence listing submitted via EFS, in compliance with 37 CFR §1.52(e)(5), is incorporated herein by reference. The sequence listing text file submitted via EFS contains the file “SequenceListing6USCNT3,” created on Feb. 17, 2010, which is 341,760 bytes in size. 
     FIELD OF THE INVENTION 
     The present invention relates to compounds, compositions, and methods for the study, diagnosis, and treatment of traits, diseases and conditions that respond to the modulation of protein tyrosine phosphatase-1B (PTP-1B) gene expression and/or activity. The present invention is also directed to compounds, compositions, and methods relating to traits, diseases and conditions that respond to the modulation of expression and/or activity of genes involved in protein tyrosine phosphatase-1B (PTP-1B) gene expression pathways or other cellular processes that mediate the maintenance or development of such traits, diseases and conditions. Specifically, the invention relates to small nucleic acid molecules, such as short interfering nucleic acid (siNA), short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), and short hairpin RNA (shRNA) molecules capable of mediating or that mediate RNA interference (RNAi) against protein tyrosine phosphatase-1B (PTP-1B) gene expression. Such small nucleic acid molecules are useful, for example, in providing compositions for treatment of traits, diseases and conditions that can respond to modulation of PTP-1B expression in a subject, such as obesity, insulin resistance, diabetes (eg. type II and type I diabetes) and any other disease, condition, trait or indication that can respond to the level of PTP-1B in a cell or tissue. 
     BACKGROUND OF THE INVENTION 
     The following is a discussion of relevant art pertaining to RNAi. The discussion is provided only for understanding of the invention that follows. The summary is not an admission that any of the work described below is prior art to the claimed invention. 
     RNA interference refers to the process of sequence-specific post-transcriptional gene silencing in animals mediated by short interfering RNAs (siRNAs) (Zamore et al., 2000, Cell, 101, 25-33; Fire et al., 1998,  Nature,  391, 806; Hamilton et al., 1999,  Science,  286, 950-951; Lin et al., 1999,  Nature,  402, 128-129; Sharp, 1999, Genes &amp; Dev., 13:139-141; and Strauss, 1999,  Science,  286, 886). The corresponding process in plants (Heifetz et al., International PCT Publication No. WO 99/61631) is commonly referred to as post-transcriptional gene silencing or RNA silencing and is also referred to as quelling in fungi. The process of post-transcriptional gene silencing is thought to be an evolutionarily-conserved cellular defense mechanism used to prevent the expression of foreign genes and is commonly shared by diverse flora and phyla (Fire et al., 1999,  Trends Genet.,  15, 358). Such protection from foreign gene expression may have evolved in response to the production of double-stranded RNAs (dsRNAs) derived from viral infection or from the random integration of transposon elements into a host genome via a cellular response that specifically destroys homologous single-stranded RNA or viral genomic RNA. The presence of dsRNA in cells triggers the RNAi response through a mechanism that has yet to be fully characterized. This mechanism appears to be different from other known mechanisms involving double stranded RNA-specific ribonucleases, such as the interferon response that results from dsRNA-mediated activation of protein kinase PKR and 2′,5′-oligoadenylate synthetase resulting in non-specific cleavage of mRNA by ribonuclease L (see for example U.S. Pat. Nos. 6,107,094; 5,898,031; Clemens et al., 1997,  J. Interferon  &amp;  Cytokine Res.,  17, 503-524; Adah et al., 2001,  Curr. Med. Chem.,  8, 1189). 
     The presence of long dsRNAs in cells stimulates the activity of a ribonuclease III enzyme referred to as dicer (Bass, 2000, Cell, 101, 235; Zamore et al., 2000, Cell, 101, 25-33; Hammond et al., 2000, Nature, 404, 293). Dicer is involved in the processing of the dsRNA into short pieces of dsRNA known as short interfering RNAs (siRNAs) (Zamore et al., 2000, Cell, 101, 25-33; Bass, 2000, Cell, 101, 235; Berstein et al., 2001,  Nature,  409, 363). Short interfering RNAs derived from dicer activity are typically about 21 to about 23 nucleotides in length and comprise about 19 base pair duplexes (Zamore et al., 2000, Cell, 101, 25-33; Elbashir et al., 2001,  Genes Dev.,  15, 188). Dicer has also been implicated in the excision of 21- and 22-nucleotide small temporal RNAs (stRNAs) from precursor RNA of conserved structure that are implicated in translational control (Hutvagner et al., 2001,  Science,  293, 834). The RNAi response also features an endonuclease complex, commonly referred to as an RNA-induced silencing complex (RISC), which mediates cleavage of single-stranded RNA having sequence complementary to the antisense strand of the siRNA duplex. Cleavage of the target RNA takes place in the middle of the region complementary to the antisense strand of the siRNA duplex (Elbashir et al., 2001,  Genes Dev.,  15, 188). 
     RNAi has been studied in a variety of systems. Fire et al., 1998,  Nature,  391, 806, were the first to observe RNAi in  C. elegans . Bahramian and Zarbl, 1999,  Molecular and Cellular Biology,  19, 274-283 and Wianny and Goetz, 1999,  Nature Cell Biol.,  2, 70, describe RNAi mediated by dsRNA in mammalian systems. Hammond et al., 2000,  Nature,  404, 293, describe RNAi in  Drosophila  cells transfected with dsRNA. Elbashir et al., 2001,  Nature,  411, 494 and Tuschl et al., International PCT Publication No. WO 01/75164, describe RNAi induced by introduction of duplexes of synthetic 21-nucleotide RNAs in cultured mammalian cells including human embryonic kidney and HeLa cells. Recent work in  Drosophila  embryonic lysates (Elbashir et al., 2001,  EMBO J.,  20, 6877 and Tuschl et al., International PCT Publication No. WO 01/75164) has revealed certain requirements for siRNA length, structure, chemical composition, and sequence that are essential to mediate efficient RNAi activity. These studies have shown that 21-nucleotide siRNA duplexes are most active when containing 3′-terminal dinucleotide overhangs. Furthermore, complete substitution of one or both siRNA strands with 2′-deoxy (2′-H) or 2′-O-methyl nucleotides abolishes RNAi activity, whereas substitution of the 3′-terminal siRNA overhang nucleotides with 2′-deoxy nucleotides (2′-H) was shown to be tolerated. Single mismatch sequences in the center of the siRNA duplex were also shown to abolish RNAi activity. In addition, these studies also indicate that the position of the cleavage site in the target RNA is defined by the 5′-end of the siRNA guide sequence rather than the 3′-end of the guide sequence (Elbashir et al., 2001,  EMBO J.,  20, 6877). Other studies have indicated that a 5′-phosphate on the target-complementary strand of a siRNA duplex is required for siRNA activity and that ATP is utilized to maintain the 5′-phosphate moiety on the siRNA (Nykanen et al., 2001,  Cell,  107, 309). 
     Studies have shown that replacing the 3′-terminal nucleotide overhanging segments of a 21-mer siRNA duplex having two-nucleotide 3′-overhangs with deoxyribonucleotides does not have an adverse effect on RNAi activity. Replacing up to four nucleotides on each end of the siRNA with deoxyribonucleotides has been reported to be well tolerated, whereas complete substitution with deoxyribonucleotides results in no RNAi activity (Elbashir et al., 2001,  EMBO J.,  20, 6877 and Tuschl et al., International PCT Publication No. WO 01/75164). In addition, Elbashir et al., supra, also report that substitution of siRNA with 2′-O-methyl nucleotides completely abolishes RNAi activity. Li et al., International PCT Publication No. WO 00/44914, and Beach et al., International PCT Publication No. WO 01/68836 preliminarily suggest that siRNA may include modifications to either the phosphate-sugar backbone or the nucleoside to include at least one of a nitrogen or sulfur heteroatom, however, neither application postulates to what extent such modifications would be tolerated in siRNA molecules, nor provides any further guidance or examples of such modified siRNA. Kreutzer et al., Canadian Patent Application No. 2,359,180, also describe certain chemical modifications for use in dsRNA constructs in order to counteract activation of double-stranded RNA-dependent protein kinase PKR, specifically 2′-amino or 2′-O-methyl nucleotides, and nucleotides containing a 2′-O or 4′-C methylene bridge. However, Kreutzer et al. similarly fails to provide examples or guidance as to what extent these modifications would be tolerated in dsRNA molecules. 
     Parrish et al., 2000,  Molecular Cell,  6, 1077-1087, tested certain chemical modifications targeting the unc-22 gene in  C. elegans  using long (&gt;25 nt) siRNA transcripts. The authors describe the introduction of thiophosphate residues into these siRNA transcripts by incorporating thiophosphate nucleotide analogs with T7 and T3 RNA polymerase and observed that RNAs with two phosphorothioate modified bases also had substantial decreases in effectiveness as RNAi. Further, Parrish et al. reported that phosphorothioate modification of more than two residues greatly destabilized the RNAs in vitro such that interference activities could not be assayed. Id. at 1081. The authors also tested certain modifications at the 2′-position of the nucleotide sugar in the long siRNA transcripts and found that substituting deoxynucleotides for ribonucleotides produced a substantial decrease in interference activity, especially in the case of Uridine to Thymidine and/or Cytidine to deoxy-Cytidine substitutions. Id. In addition, the authors tested certain base modifications, including substituting, in sense and antisense strands of the siRNA, 4-thiouracil, 5-bromouracil, 5-iodouracil, and 3-(aminoallyl)uracil for uracil, and inosine for guanosine. Whereas 4-thiouracil and 5-bromouracil substitution appeared to be tolerated, Parrish reported that inosine produced a substantial decrease in interference activity when incorporated in either strand. Parrish also reported that incorporation of 5-iodouracil and 3-(aminoallyl)uracil in the antisense strand resulted in a substantial decrease in RNAi activity as well. 
     The use of longer dsRNA has been described. For example, Beach et al., International PCT Publication No. WO 01/68836, describes specific methods for attenuating gene expression using endogenously-derived dsRNA. Tuschl et al., International PCT Publication No. WO 01/75164, describe a  Drosophila  in vitro RNAi system and the use of specific siRNA molecules for certain functional genomic and certain therapeutic applications; although Tuschl, 2001,  Chem. Biochem.,  2, 239-245, doubts that RNAi can be used to cure genetic diseases or viral infection due to the danger of activating interferon response. Li et al., International PCT Publication No. WO 00/44914, describe the use of specific long (141 bp-488 bp) enzymatically synthesized or vector expressed dsRNAs for attenuating the expression of certain target genes. Zernicka-Goetz et al., International PCT Publication No. WO 01/36646, describe certain methods for inhibiting the expression of particular genes in mammalian cells using certain long (550 bp-714 bp), enzymatically synthesized or vector expressed dsRNA molecules. Fire et al., International PCT Publication No. WO 99/32619, describe particular methods for introducing certain long dsRNA molecules into cells for use in inhibiting gene expression in nematodes. Plaetinck et al., International PCT Publication No. WO 00/01846, describe certain methods for identifying specific genes responsible for conferring a particular phenotype in a cell using specific long dsRNA molecules. Mello et al., International PCT Publication No. WO 01/29058, describe the identification of specific genes involved in dsRNA-mediated RNAi. Pachuck et al., International PCT Publication No. WO 00/63364, describe certain long (at least 200 nucleotide) dsRNA constructs. Deschamps Depaillette et al., International PCT Publication No. WO 99/07409, describe specific compositions consisting of particular dsRNA molecules combined with certain anti-viral agents. Waterhouse et al., International PCT Publication No. 99/53050 and 1998,  PNAS,  95, 13959-13964, describe certain methods for decreasing the phenotypic expression of a nucleic acid in plant cells using certain dsRNAs. Driscoll et al., International PCT Publication No. WO 01/49844, describe specific DNA expression constructs for use in facilitating gene silencing in targeted organisms. 
     Others have reported on various RNAi and gene-silencing systems. For example, Parrish et al., 2000,  Molecular Cell,  6, 1077-1087, describe specific chemically-modified dsRNA constructs targeting the unc-22 gene of  C. elegans . Grossniklaus, International PCT Publication No. WO 01/38551, describes certain methods for regulating polycomb gene expression in plants using certain dsRNAs. Churikov et al., International PCT Publication No. WO 01/42443, describe certain methods for modifying genetic characteristics of an organism using certain dsRNAs. Cogoni et al, International PCT Publication No. WO 01/53475, describe certain methods for isolating a Neurospora silencing gene and uses thereof. Reed et al., International PCT Publication No. WO 01/68836, describe certain methods for gene silencing in plants. Honer et al., International PCT Publication No. WO 01/70944, describe certain methods of drug screening using transgenic nematodes as Parkinson&#39;s Disease models using certain dsRNAs. Deak et al., International PCT Publication No. WO 01/72774, describe certain  Drosophila -derived gene products that may be related to RNAi in  Drosophila . Arndt et al., International PCT Publication No. WO 01/92513 describe certain methods for mediating gene suppression by using factors that enhance RNAi. Tuschl et al., International PCT Publication No. WO 02/44321, describe certain synthetic siRNA constructs. Pachuk et al., International PCT Publication No. WO 00/63364, and Satishchandran et al., International PCT Publication No. WO 01/04313, describe certain methods and compositions for inhibiting the function of certain polynucleotide sequences using certain long (over 250 bp), vector expressed dsRNAs. Echeverri et al., International PCT Publication No. WO 02/38805, describe certain  C. elegans  genes identified via RNAi. Kreutzer et al., International PCT Publications Nos. WO 02/055692, WO 02/055693, and EP 1144623 B1 describes certain methods for inhibiting gene expression using dsRNA. Graham et al., International PCT Publications Nos. WO 99/49029 and WO 01/70949, and AU 4037501 describe certain vector expressed siRNA molecules. Fire et al., U.S. Pat. No. 6,506,559, describe certain methods for inhibiting gene expression in vitro using certain long dsRNA (299 bp-1033 bp) constructs that mediate RNAi. Martinez et al., 2002,  Cell,  110, 563-574, describe certain single stranded siRNA constructs, including certain 5′-phosphorylated single stranded siRNAs that mediate RNA interference in Hela cells. Harborth et al., 2003, Antisense &amp; Nucleic Acid Drug Development, 13, 83-105, describe certain chemically and structurally modified siRNA molecules. Chiu and Rana, 2003, RNA, 9, 1034-1048, describe certain chemically and structurally modified siRNA molecules. Woolf et al., International PCT Publication Nos. WO 03/064626 and WO 03/064625 describe certain chemically modified dsRNA constructs. McSwiggen et al., International PCT Publication No. WO 01/16312, describes nucleic acid modulators of PTP-1B. 
     SUMMARY OF THE INVENTION 
     This invention relates to compounds, compositions, and methods useful for modulating protein tyrosine phosphatase-1B (PTP-1B) gene expression using short interfering nucleic acid (siNA) molecules. This invention also relates to compounds, compositions, and methods useful for modulating the expression and activity of other genes involved in pathways of protein tyrosine phosphatase-1B (PTP-1B) gene expression and/or activity by RNA interference (RNAi) using small nucleic acid molecules. In particular, the instant invention features small nucleic acid molecules, such as short interfering nucleic acid (siNA), short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), and short hairpin RNA (shRNA) molecules and methods used to modulate the expression protein tyrosine phosphatase-1B (PTP-1B) genes. 
     A siNA of the invention can be unmodified or chemically-modified. A siNA of the instant invention can be chemically synthesized, expressed from a vector or enzymatically synthesized. The instant invention also features various chemically-modified synthetic short interfering nucleic acid (siNA) molecules capable of modulating PTP-1B gene expression or activity in cells by RNA interference (RNAi). The use of chemically-modified siNA improves various properties of native siNA molecules through increased resistance to nuclease degradation in vivo and/or through improved cellular uptake. Further, contrary to earlier published studies, siNA having multiple chemical modifications retains its RNAi activity. The siNA molecules of the instant invention provide useful reagents and methods for a variety of therapeutic, cosmetic, veterinary, diagnostic, target validation, genomic discovery, genetic engineering, and pharmacogenomic applications. 
     In one embodiment, the invention features one or more siNA molecules and methods that independently or in combination modulate the expression of PTP-1B genes en coding proteins, such as proteins comprising PTP-1B associated with the maintenance and/or development of diabetes (e.g., type 1 and type 2), obesity, and/or insulin resistance in a subject or organism such as genes encoding sequences comprising those sequences referred to by GenBank Accession Nos. shown in Table I, referred to herein generally as PTP-1B (also known as PTPN1). The description below of the various aspects and embodiments of the invention is provided with reference to exemplary PTP-1B gene. However, the various aspects and embodiments are also directed to other PTP-1B genes, such as PTP-1B homolog genes and transcript variants and polymorphisms (e.g., single nucleotide polymorphism, (SNPs)) associated with certain PTP-1B genes. As such, the various aspects and embodiments are also directed to other genes that are involved in PTP-1B mediated pathways of signal transduction or gene expression that are involved, for example, in the maintenance and/or development of conditions or disease states such as diabetes (e.g., type 1 and type 2), obesity, and/or insulin resistance in a subject or organism. These additional genes can be analyzed for target sites using the methods described for PTP-1B genes herein. Thus, the modulation of other genes and the effects of such modulation of the other genes can be performed, determined, and measured as described herein. 
     In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that down-regulates expression of a PTP-1B gene, wherein said siNA molecule comprises about 15 to about 28 base pairs. 
     In one embodiment, the invention features a double stranded short interfering nucleic acid (siNA) molecule that directs cleavage of a PTP-1B RNA via RNA interference (RNAi), wherein the double stranded siNA molecule comprises a first and a second strand, each strand of the siNA molecule is about 18 to about 28 nucleotides in length, the first strand of the siNA molecule comprises nucleotide sequence having sufficient complementarity to the PTP-1B RNA for the siNA molecule to direct cleavage of the PTP-1B RNA via RNA interference, and the second strand of said siNA molecule comprises nucleotide sequence that is complementary to the first strand. 
     In one embodiment, the invention features a double stranded short interfering nucleic acid (siNA) molecule that directs cleavage of a PTP-1B RNA via RNA interference (RNAi), wherein the double stranded siNA molecule comprises a first and a second strand, each strand of the siNA molecule is about 18 to about 23 nucleotides in length, the first strand of the siNA molecule comprises nucleotide sequence having sufficient complementarity to the PTP-1B RNA for the siNA molecule to direct cleavage of the PTP-1B RNA via RNA interference, and the second strand of said siNA molecule comprises nucleotide sequence that is complementary to the first strand. 
     In one embodiment, the invention features a chemically synthesized double stranded short interfering nucleic acid (siNA) molecule that directs cleavage of a PTP-1B RNA via RNA interference (RNAi), wherein each strand of the siNA molecule is about 18 to about 28 nucleotides in length; and one strand of the siNA molecule comprises nucleotide sequence having sufficient complementarity to the PTP-1B RNA for the siNA molecule to direct cleavage of the PTP-1B RNA via RNA interference. 
     In one embodiment, the invention features a chemically synthesized double stranded short interfering nucleic acid (siNA) molecule that directs cleavage of a PTP-1B RNA via RNA interference (RNAi), wherein each strand of the siNA molecule is about 18 to about 23 nucleotides in length; and one strand of the siNA molecule comprises nucleotide sequence having sufficient complementarity to the PTP-1 B RNA for the siNA molecule to direct cleavage of the PTP-1B RNA via RNA interference. 
     In one embodiment, the invention features a siNA molecule that down-regulates expression of a PTP-1B gene or that directs cleavage of a PTP-1B RNA, for example, wherein the PTP-1B gene or RNA comprises PTP-1B encoding sequence. In one embodiment, the invention features a siNA molecule that down-regulates expression of a PTP-1B gene or that directs cleavage of a PTP-1B RNA, for example, wherein the PTP-1B gene or RNA comprises PTP-1B non-coding sequence or regulatory elements involved in PTP-1B gene expression. 
     In one embodiment, a siNA of the invention is used to inhibit the expression of PTP-1B genes or a PTP-1B gene family, wherein the genes or gene family sequences share sequence homology. Such homologous sequences can be identified as is known in the art, for example using sequence alignments. siNA molecules can be designed to target such homologous sequences, for example using perfectly complementary sequences or by incorporating non-canonical base pairs, for example mismatches and/or wobble base pairs, that can provide additional target sequences. In instances where mismatches are identified, non-canonical base pairs (for example, mismatches and/or wobble bases) can be used to generate siNA molecules that target more than one gene sequence. In a non-limiting example, non-canonical base pairs such as UU and CC base pairs are used to generate siNA molecules that are capable of targeting sequences for differing PTP-1B targets that share sequence homology. As such, one advantage of using siNAs of the invention is that a single siNA can be designed to include nucleic acid sequence that is complementary to the nucleotide sequence that is conserved between the homologous genes. In this approach, a single siNA can be used to inhibit expression of more than one gene instead of using more than one siNA molecule to target the different genes. 
     In one embodiment, the invention features a siNA molecule having RNAi activity against PTP-1B RNA, wherein the siNA molecule comprises a sequence complementary to any RNA having PTP-1B encoding sequence, such as those sequences having GenBank Accession Nos. shown in Table I. In another embodiment, the invention features a siNA molecule having RNAi activity against PTP-1B RNA, wherein the siNA molecule comprises a sequence complementary to an RNA having variant PTP-1B encoding sequence, for example other mutant PTP-1B genes not shown in Table I but known in the art to be associated with the maintenance and/or development of diabetes (e.g., type 1 and type 2), obesity, and/or insulin resistance in a subject or organism. Chemical modifications as shown in Tables III and IV or otherwise described herein can be applied to any siNA construct of the invention. In another embodiment, a siNA molecule of the invention includes a nucleotide sequence that can interact with nucleotide sequence of a PTP-1B gene and thereby mediate silencing of PTP-1B gene expression, for example, wherein the siNA mediates regulation of PTP-1B gene expression by cellular processes that modulate the chromatin structure or methylation patterns of the PTP-1B gene and prevent transcription of the PTP-1B gene. 
     In one embodiment, siNA molecules of the invention are used to down regulate or inhibit the expression of proteins arising from PTP-1B haplotype polymorphisms that are associated with a trait, disease or condition such as diabetes (e.g., type 1 and type 2), obesity, and/or insulin resistance in a subject or organism. Analysis of genes, or protein or RNA levels can be used to identify subjects with such polymorphisms or those subjects who are at risk of developing traits, conditions, or diseases described herein. These subjects are amenable to treatment, for example, treatment with siNA molecules of the invention and any other composition useful in treating diseases related to PTP-1B gene expression. As such, analysis of PTP-1B protein or RNA levels can be used to determine treatment type and the course of therapy in treating a subject. Monitoring of PTP-1B protein or RNA levels can be used to predict treatment outcome and to determine the efficacy of compounds and compositions that modulate the level and/or activity of certain PTP-1B proteins associated with a trait, condition, or disease. 
     In one embodiment of the invention a siNA molecule comprises an antisense strand comprising a nucleotide sequence that is complementary to a nucleotide sequence or a portion thereof encoding a PTP-1B protein. The siNA further comprises a sense strand, wherein said sense strand comprises a nucleotide sequence of a PTP-1B gene or a portion thereof. 
     In another embodiment, a siNA molecule comprises an antisense region comprising a nucleotide sequence that is complementary to a nucleotide sequence encoding a PTP-1B protein or a portion thereof The siNA molecule further comprises a sense region, wherein said sense region comprises a nucleotide sequence of a PTP-1B gene or a portion thereof. 
     In another embodiment, the invention features a siNA molecule comprising nucleotide sequence, for example, nucleotide sequence in the antisense region of the siNA molecule that is complementary to a nucleotide sequence or portion of sequence of a PTP-1B gene. In another embodiment, the invention features a siNA molecule comprising a region, for example, the antisense region of the siNA construct, complementary to a sequence comprising a PTP-1B gene sequence or a portion thereof. 
     In one embodiment, the antisense region of siNA constructs comprises a sequence complementary to sequence having any of target SEQ ID NOs. shown in Tables II and III. In one embodiment, the antisense region of siNA constructs of the invention constructs comprises sequence having any of antisense (lower) SEQ ID NOs. in Tables II and III and  FIGS. 4 and 5 . In another embodiment, the sense region of siNA constructs of the invention comprises sequence having any of sense (upper) SEQ ID NOs. in Tables II and III and  FIGS. 4 and 5 . 
     In one embodiment, a siNA molecule of the invention comprises any of SEQ ID NOs. 1-755. The sequences shown in SEQ ID NOs: 1-755 are not limiting. A siNA molecule of the invention can comprise any contiguous PTP-1B sequence (e.g., about 15 to about 25 or more, or about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 or more contiguous PTP-1B nucleotides). 
     In yet another embodiment, the invention features a siNA molecule comprising a sequence, for example, the antisense sequence of the siNA construct, complementary to a sequence or portion of sequence comprising sequence represented by GenBank Accession Nos. shown in Table I. Chemical modifications in Tables III and IV and described herein can be applied to any siNA construct of the invention. 
     In one embodiment of the invention a siNA molecule comprises an antisense strand having about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides, wherein the antisense strand is complementary to a RNA sequence or a portion thereof encoding PTP-1B, and wherein said siNA further comprises a sense strand having about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides, and wherein said sense strand and said antisense strand are distinct nucleotide sequences where at least about 15 nucleotides in each strand are complementary to the other strand. 
     In another embodiment of the invention a siNA molecule of the invention comprises an antisense region having about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides, wherein the antisense region is complementary to a RNA sequence encoding PTP-1B, and wherein said siNA further comprises a sense region having about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides, wherein said sense region and said antisense region are comprised in a linear molecule where the sense region comprises at least about 15 nucleotides that are complementary to the antisense region. 
     In one embodiment, a siNA molecule of the invention has RNAi activity that modulates expression of RNA encoded by a PTP-1B gene. Because PTP-1B genes can share some degree of sequence homology with each other, siNA molecules can be designed to target a class of PTP-1B genes or alternately specific PTP-1B genes (e.g., polymorphic variants) by selecting sequences that are either shared amongst different PTP-1B targets or alternatively that are unique for a specific PTP-1B target. Therefore, in one embodiment, the siNA molecule can be designed to target conserved regions of PTP-1B RNA sequences having homology among several PTP-1B gene variants so as to target a class of PTP-1B genes with one siNA molecule. Accordingly, in one embodiment, the siNA molecule of the invention modulates the expression of one or both PTP-1B alleles in a subject. In another embodiment, the siNA molecule can be designed to target a sequence that is unique to a specific PTP-1B RNA sequence (e.g., a single PTP-1B allele or PTP-1B single nucleotide polymorphism (SNP)) due to the high degree of specificity that the siNA molecule requires to mediate RNAi activity. 
     In one embodiment, nucleic acid molecules of the invention that act as mediators of the RNA interference gene silencing response are double-stranded nucleic acid molecules. In another embodiment, the siNA molecules of the invention consist of duplex nucleic acid molecules containing about 15 to about 30 base pairs between oligonucleotides comprising about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides. In yet another embodiment, siNA molecules of the invention comprise duplex nucleic acid molecules with overhanging ends of about 1 to about 3 (e.g., about 1, 2, or 3) nucleotides, for example, about 21-nucleotide duplexes with about 19 base pairs and 3′-terminal mononucleotide, dinucleotide, or trinucleotide overhangs. In yet another embodiment, siNA molecules of the invention comprise duplex nucleic acid molecules with blunt ends, where both ends are blunt, or alternatively, where one of the ends is blunt. 
     In one embodiment, the invention features one or more chemically-modified siNA constructs having specificity for PTP-1B expressing nucleic acid molecules, such as RNA encoding a PTP-1B protein or non-coding RNA associated with the expression of PTP-1B genes. In one embodiment, the invention features a RNA based siNA molecule (e.g., a siNA comprising 2′-OH nucleotides) having specificity for PTP-1B expressing nucleic acid molecules that includes one or more chemical modifications described herein. Non-limiting examples of such chemical modifications include without limitation phosphorothioate internucleotide linkages, 2′-deoxyribonucleotides, 2′-O-methyl ribonucleotides, 2′-deoxy-2′-fluoro ribonucleotides, 4′-thio ribonucleotides, 2′-O-trifluoromethyl nucleotides, 2′-O-ethyl-trifluoromethoxy nucleotides, 2′-O-difluoromethoxy-ethoxy nucleotides (see for example U.S. Ser. No. 10/981,966 filed Nov. 5, 2004, incorporated by reference herein), “universal base” nucleotides, “acyclic” nucleotides, 5-C-methyl nucleotides, and terminal glyceryl and/or inverted deoxy abasic residue incorporation. These chemical modifications, when used in various siNA constructs, (e.g., RNA based siNA constructs), are shown to preserve RNAi activity in cells while at the same time, dramatically increasing the serum stability of these compounds. Furthermore, contrary to the data published by Parrish et al., supra, applicant demonstrates that multiple (greater than one) phosphorothioate substitutions are well-tolerated and confer substantial increases in serum stability for modified siNA constructs. 
     In one embodiment, a siNA molecule of the invention comprises modified nucleotides while maintaining the ability to mediate RNAi. The modified nucleotides can be used to improve in vitro or in vivo characteristics such as stability, activity, toxicity, immune response, and/or bioavailability. For example, a siNA molecule of the invention can comprise modified nucleotides as a percentage of the total number of nucleotides present in the siNA molecule. As such, a siNA molecule of the invention can generally comprise about 5% to about 100% modified nucleotides (e.g., about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% modified nucleotides). The actual percentage of modified nucleotides present in a given siNA molecule will depend on the total number of nucleotides present in the siNA. If the siNA molecule is single stranded, the percent modification can be based upon the total number of nucleotides present in the single stranded siNA molecules. Likewise, if the siNA molecule is double stranded, the percent modification can be based upon the total number of nucleotides present in the sense strand, antisense strand, or both the sense and antisense strands. 
     A siNA molecule of the invention can comprise modified nucleotides at various locations within the siNA molecule. In one embodiment, a double stranded siNA molecule of the invention comprises modified nucleotides at internal base paired positions within the siNA duplex. For example, internal positions can comprise positions from about 3 to about 19 nucleotides from the 5′-end of either sense or antisense strand or region of a 21 nucleotide siNA duplex having 19 base pairs and two nucleotide 3′-overhangs. In another embodiment, a double stranded siNA molecule of the invention comprises modified nucleotides at non-base paired or overhang regions of the siNA molecule. For example, overhang positions can comprise positions from about 20 to about 21 nucleotides from the 5′-end of either sense or antisense strand or region of a 21 nucleotide siNA duplex having 19 base pairs and two nucleotide 3′-overhangs. In another embodiment, a double stranded siNA molecule of the invention comprises modified nucleotides at terminal positions of the siNA molecule. For example, such terminal regions include the 3′-position, 5′-position, for both 3′ and 5′-positions of the sense and/or antisense strand or region of the siNA molecule. In another embodiment, double stranded siNA molecule of the invention comprises modified nucleotides at base-paired or internal positions, non-base paired or overhang regions, and/or terminal regions, or any combination thereof. 
     One aspect of the invention features a double-stranded short interfering nucleic acid (siNA) molecule that down-regulates expression of a PTP-1B gene or that directs cleavage of a PTP-1B RNA. In one embodiment, the double stranded siNA molecule comprises one or more chemical modifications and each strand of the double-stranded siNA is about 21 nucleotides long. In one embodiment, the double-stranded siNA molecule does not contain any ribonucleotides. In another embodiment, the double-stranded siNA molecule comprises one or more ribonucleotides. In one embodiment, each strand of the double-stranded siNA molecule independently comprises about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides, wherein each strand comprises about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides that are complementary to the nucleotides of the other strand. In one embodiment, one of the strands of the double-stranded siNA molecule comprises a nucleotide sequence that is complementary to a nucleotide sequence or a portion thereof of the PTP-1B gene, and the second strand of the double-stranded siNA molecule comprises a nucleotide sequence substantially similar to the nucleotide sequence of the PTP-1B gene or a portion thereof. 
     In another embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that down-regulates expression of a PTP-1B gene or that directs cleavage of a PTP-1B RNA, comprising an antisense region, wherein the anti sense region comprises a nucleotide sequence that is complementary to a nucleotide sequence of the PTP-1B gene or a portion thereof, and a sense region, wherein the sense region comprises a nucleotide sequence substantially similar to the nucleotide sequence of the PTP-1B gene or a portion thereof. In one embodiment, the antisense region and the sense region independently comprise about 15 to about 30 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides, wherein the antisense region comprises about 15 to about 30 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides that are complementary to nucleotides of the sense region. 
     In another embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that down-regulates expression of a PTP-1B gene or that directs cleavage of a PTP-1B RNA, comprising a sense region and an antisense region, wherein the antisense region comprises a nucleotide sequence that is complementary to a nucleotide sequence of RNA encoded by the PTP-1B gene or a portion thereof and the sense region comprises a nucleotide sequence that is complementary to the antisense region. 
     In one embodiment, a siNA molecule of the invention comprises blunt ends, i.e., ends that do not include any overhanging nucleotides. For example, a siNA molecule comprising modifications described herein (e.g., comprising nucleotides having Formulae I-VII or siNA constructs comprising “Stab 00”-“Stab 34” or “Stab 3F”-“Stab 34F” (Table IV) or any combination thereof (see Table IV)) and/or any length described herein can comprise blunt ends or ends with no overhanging nucleotides. 
     In one embodiment, any siNA molecule of the invention can comprise one or more blunt ends, i.e. where a blunt end does not have any overhanging nucleotides. In one embodiment, the blunt ended siNA molecule has a number of base pairs equal to the number of nucleotides present in each strand of the siNA molecule. In another embodiment, the siNA molecule comprises one blunt end, for example wherein the 5′-end of the antisense strand and the 3′-end of the sense strand do not have any overhanging nucleotides. In another example, the siNA molecule comprises one blunt end, for example wherein the 3′-end of the antisense strand and the 5′-end of the sense strand do not have any overhanging nucleotides. In another example, a siNA molecule comprises two blunt ends, for example wherein the 3′-end of the antisense strand and the 5′-end of the sense strand as well as the 5′-end of the antisense strand and 3′-end of the sense strand do not have any overhanging nucleotides. A blunt ended siNA molecule can comprise, for example, from about 15 to about 30 nucleotides (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides). Other nucleotides present in a blunt ended siNA molecule can comprise, for example, mismatches, bulges, loops, or wobble base pairs to modulate the activity of the siNA molecule to mediate RNA interference. 
     By “blunt ends” is meant symmetric termini or termini of a double stranded siNA molecule having no overhanging nucleotides. The two strands of a double stranded siNA molecule align with each other without over-hanging nucleotides at the termini. For example, a blunt ended siNA construct comprises terminal nucleotides that are complementary between the sense and antisense regions of the siNA molecule. 
     In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that down-regulates expression of a PTP-1B gene or that directs cleavage of a PTP-1B RNA, wherein the siNA molecule is assembled from two separate oligonucleotide fragments wherein one fragment comprises the sense region and the second fragment comprises the antisense region of the siNA molecule. The sense region can be connected to the antisense region via a linker molecule, such as a polynucleotide linker or a non-nucleotide linker. 
     In one embodiment, the invention features double-stranded short interfering nucleic acid (siNA) molecule that down-regulates expression of a PTP-1B gene or that directs cleavage of a PTP-1B RNA, wherein the siNA molecule comprises about 15 to about 30 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) base pairs, and wherein each strand of the siNA molecule comprises one or more chemical modifications. In another embodiment, one of the strands of the double-stranded siNA molecule comprises a nucleotide sequence that is complementary to a nucleotide sequence of a PTP-1B gene or a portion thereof, and the second strand of the double-stranded siNA molecule comprises a nucleotide sequence substantially similar to the nucleotide sequence or a portion thereof of the PTP-1B gene. In another embodiment, one of the strands of the double-stranded siNA molecule comprises a nucleotide sequence that is complementary to a nucleotide sequence of a PTP-1B gene or portion thereof, and the second strand of the double-stranded siNA molecule comprises a nucleotide sequence substantially similar to the nucleotide sequence or portion thereof of the PTP-1B gene. In another embodiment, each strand of the siNA molecule comprises about 15 to about 30 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides, and each strand comprises at least about 15 to about 30 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides that are complementary to the nucleotides of the other strand. The PTP-1B gene can comprise, for example, sequences referred to in Table I. 
     In one embodiment, a siNA molecule of the invention comprises no ribonucleotides. In another embodiment, a siNA molecule of the invention comprises ribonucleotides. 
     In one embodiment, a siNA molecule of the invention comprises an antisense region comprising a nucleotide sequence that is complementary to a nucleotide sequence of a PTP-1B gene or a portion thereof, and the siNA further comprises a sense region comprising a nucleotide sequence substantially similar to the nucleotide sequence of the PTP-1B gene or a portion thereof. In another embodiment, the antisense region and the sense region each comprise about 15 to about 30 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides and the antisense region comprises at least about 15 to about 30 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides that are complementary to nucleotides of the sense region. The PTP-1B gene can comprise, for example, sequences referred to in Table I. In another embodiment, the siNA is a double stranded nucleic acid molecule, where each of the two strands of the siNA molecule independently comprise about 15 to about 40 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 23, 33, 34, 35, 36, 37, 38, 39, or 40) nucleotides, and where one of the strands of the siNA molecule comprises at least about 15 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 or more) nucleotides that are complementary to the nucleic acid sequence of the PTP-1B gene or a portion thereof. 
     In one embodiment, a siNA molecule of the invention comprises a sense region and an antisense region, wherein the antisense region comprises a nucleotide sequence that is complementary to a nucleotide sequence of RNA encoded by a PTP-1B gene, or a portion thereof, and the sense region comprises a nucleotide sequence that is complementary to the antisense region. In one embodiment, the siNA molecule is assembled from two separate oligonucleotide fragments, wherein one fragment comprises the sense region and the second fragment comprises the antisense region of the siNA molecule. In another embodiment, the sense region is connected to the antisense region via a linker molecule. In another embodiment, the sense region is connected to the antisense region via a linker molecule, such as a nucleotide or non-nucleotide linker. The PTP-1B gene can comprise, for example, sequences referred in to Table I. 
     In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that down-regulates expression of a PTP-1B gene or that directs cleavage of a PTP-1B RNA, comprising a sense region and an antisense region, wherein the antisense region comprises a nucleotide sequence that is complementary to a nucleotide sequence of RNA encoded by the PTP-1 B gene or a portion thereof and the sense region comprises a nucleotide sequence that is complementary to the antisense region, and wherein the siNA molecule has one or more modified pyrimidine and/or purine nucleotides. In one embodiment, the pyrimidine nucleotides in the sense region are 2′-O-methyl pyrimidine nucleotides or 2′-deoxy-2′-fluoro pyrimidine nucleotides and the purine nucleotides present in the sense region are 2′-deoxy purine nucleotides. In another embodiment, the pyrimidine nucleotides in the sense region are T-deoxy-2′-fluoro pyrimidine nucleotides and the purine nucleotides present in the sense region are 2′-O-methyl purine nucleotides. In another embodiment, the pyrimidine nucleotides in the sense region are 2′-deoxy-2′-fluoro pyrimidine nucleotides and the purine nucleotides present in the sense region are 2′-deoxy purine nucleotides. In one embodiment, the pyrimidine nucleotides in the antisense region are 2′-deoxy-2′-fluoro pyrimidine nucleotides and the purine nucleotides present in the antisense region are 2′-O-methyl or 2′-deoxy purine nucleotides. In another embodiment of any of the above-described siNA molecules, any nucleotides present in a non-complementary region of the sense strand (e.g. overhang region) are 2′-deoxy nucleotides. 
     In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that down-regulates expression of a PTP-1B gene or that directs cleavage of a PTP-1B RNA, wherein the siNA molecule is assembled from two separate oligonucleotide fragments wherein one fragment comprises the sense region and the second fragment comprises the antisense region of the siNA molecule, and wherein the fragment comprising the sense region includes a terminal cap moiety at the 5′-end, the 3′-end, or both of the 5′ and 3′ ends of the fragment. In one embodiment, the terminal cap moiety is an inverted deoxy abasic moiety or glyceryl moiety. In one embodiment, each of the two fragments of the siNA molecule independently comprise about 15 to about 30 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides. In another embodiment, each of the two fragments of the siNA molecule independently comprise about 15 to about 40 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 23, 33, 34, 35, 36, 37, 38, 39, or 40) nucleotides. In a non-limiting example, each of the two fragments of the siNA molecule comprise about 21 nucleotides. 
     In one embodiment, the invention features a siNA molecule comprising at least one modified nucleotide, wherein the modified nucleotide is a 2′-deoxy-2′-fluoro nucleotide, 2′-O-trifluoromethyl nucleotide, 2′-O-ethyl-trifluoromethoxy nucleotide, or 2′-O-difluoromethoxy-ethoxy nucleotide or any other modified nucleoside/nucleotide described in U.S. Ser. No. 10/981,966 filed Nov. 5, 2004, incorporated by reference herein. The siNA can be, for example, about 15 to about 40 nucleotides in length. In one embodiment, all pyrimidine nucleotides present in the siNA are 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy, 4′-thio pyrimidine nucleotides. In one embodiment, the modified nucleotides in the siNA include at least one 2′-deoxy-2′-fluoro cytidine or 2′-deoxy-2′-fluoro uridine nucleotide. In another embodiment, the modified nucleotides in the siNA include at least one 2′-fluoro cytidine and at least one 2′-deoxy-2′-fluoro uridine nucleotides. In one embodiment, all uridine nucleotides present in the siNA are 2′-deoxy-2′-fluoro uridine nucleotides. In one embodiment, all cytidine nucleotides present in the siNA are 2′-deoxy-2′-fluoro cytidine nucleotides. In one embodiment, all adenosine nucleotides present in the siNA are 2′-deoxy-2′-fluoro adenosine nucleotides. In one embodiment, all guanosine nucleotides present in the siNA are 2′-deoxy-2′-fluoro guanosine nucleotides. The siNA can further comprise at least one modified internucleotidic linkage, such as phosphorothioate linkage. In one embodiment, the 2′-deoxy-2′-fluoronucleotides are present at specifically selected locations in the siNA that are sensitive to cleavage by ribonucleases, such as locations having pyrimidine nucleotides. 
     In one embodiment, the invention features a method of increasing the stability of a siNA molecule against cleavage by ribonucleases comprising introducing at least one modified nucleotide into the siNA molecule, wherein the modified nucleotide is a 2′-deoxy-2′-fluoro nucleotide. In one embodiment, all pyrimidine nucleotides present in the siNA are 2′-deoxy-2′-fluoro pyrimidine nucleotides. In one embodiment, the modified nucleotides in the siNA include at least one 2′-deoxy-2′-fluoro cytidine or 2′-deoxy-2′-fluoro uridine nucleotide. In another embodiment, the modified nucleotides in the siNA include at least one 2′-fluoro cytidine and at least one 2′-deoxy-2′-fluoro uridine nucleotides. In one embodiment, all uridine nucleotides present in the siNA are 2′-deoxy-2′-fluoro uridine nucleotides. In one embodiment, all cytidine nucleotides present in the siNA are 2′-deoxy-2′-fluoro cytidine nucleotides. In one embodiment, all adenosine nucleotides present in the siNA are 2′-deoxy-2′-fluoro adenosine nucleotides. In one embodiment, all guanosine nucleotides present in the siNA are 2′-deoxy-2′-fluoro guanosine nucleotides. The siNA can further comprise at least one modified internucleotidic linkage, such as a phosphorothioate linkage. In one embodiment, the 2′-deoxy-2′-fluoronucleotides are present at specifically selected locations in the siNA that are sensitive to cleavage by ribonucleases, such as locations having pyrimidine nucleotides. 
     In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that down-regulates expression of a PTP-1B gene or that directs cleavage of a PTP-1B RNA, comprising a sense region and an antisense region, wherein the antisense region comprises a nucleotide sequence that is complementary to a nucleotide sequence of RNA encoded by the PTP-1B gene or a portion thereof and the sense region comprises a nucleotide sequence that is complementary to the anti sense region, and wherein the purine nucleotides present in the antisense region comprise 2′-deoxy-purine nucleotides. In an alternative embodiment, the purine nucleotides present in the antisense region comprise 2′-O-methyl purine nucleotides. In either of the above embodiments, the antisense region can comprise a phosphorothioate internucleotide linkage at the 3′ end of the antisense region. Alternatively, in either of the above embodiments, the antisense region can comprise a glyceryl modification at the 3′ end of the antisense region. In another embodiment of any of the above-described siNA molecules, any nucleotides present in a non-complementary region of the antisense strand (e.g. overhang region) are 2′-deoxy nucleotides. 
     In one embodiment, the antisense region of a siNA molecule of the invention comprises sequence complementary to a portion of an endogenous transcript having sequence unique to a particular PTP-1B disease or trait related allele in a subject or organism, such as sequence comprising a single nucleotide polymorphism (SNP) associated with the disease or trait specific allele. As such, the antisense region of a siNA molecule of the invention can comprise sequence complementary to sequences that are unique to a particular allele to provide specificity in mediating selective RNAi against the disease, condition, or trait related allele. 
     In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that down-regulates expression of a PTP-1B gene or that directs cleavage of a PTP-1B RNA, wherein the siNA molecule is assembled from two separate oligonucleotide fragments wherein one fragment comprises the sense region and the second fragment comprises the antisense region of the siNA molecule. In another embodiment, the siNA molecule is a double stranded nucleic acid molecule, where each strand is about 21 nucleotides long and where about 19 nucleotides of each fragment of the siNA molecule are base-paired to the complementary nucleotides of the other fragment of the siNA molecule, wherein at least two 3′ terminal nucleotides of each fragment of the siNA molecule are not base-paired to the nucleotides of the other fragment of the siNA molecule. In another embodiment, the siNA molecule is a double stranded nucleic acid molecule, where each strand is about 19 nucleotide long and where the nucleotides of each fragment of the siNA molecule are base-paired to the complementary nucleotides of the other fragment of the siNA molecule to form at least about 15 (e.g., 15, 16, 17, 18, or 19) base pairs, wherein one or both ends of the siNA molecule are blunt ends. In one embodiment, each of the two 3′ terminal nucleotides of each fragment of the siNA molecule is a 2′-deoxy-pyrimidine nucleotide, such as a 2′-deoxy-thymidine. In another embodiment, all nucleotides of each fragment of the siNA molecule are base-paired to the complementary nucleotides of the other fragment of the siNA molecule. In another embodiment, the siNA molecule is a double stranded nucleic acid molecule of about 19 to about 25 base pairs having a sense region and an antisense region, where about 19 nucleotides of the antisense region are base-paired to the nucleotide sequence or a portion thereof of the RNA encoded by the PTP-1B gene. In another embodiment, about 21 nucleotides of the antisense region are base-paired to the nucleotide sequence or a portion thereof of the RNA encoded by the PTP-1B gene. In any of the above embodiments, the 5′-end of the fragment comprising said antisense region can optionally include a phosphate group. 
     In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that inhibits the expression of a PTP-1B RNA sequence (e.g., wherein said target RNA sequence is encoded by a PTP-1B gene involved in the PTP-1B pathway), wherein the siNA molecule does not contain any ribonucleotides and wherein each strand of the double-stranded siNA molecule is about 15 to about 30 nucleotides. In one embodiment, the siNA molecule is 21 nucleotides in length. Examples of non-ribonucleotide containing siNA constructs are combinations of stabilization chemistries shown in Table IV in any combination of Sense/Antisense chemistries, such as Stab 7/8, Stab 7/11, Stab 8/8, Stab 18/8, Stab 18/11, Stab 12/13, Stab 7/13, Stab 18/13, Stab 7/19, Stab 8/19, Stab 18/19, Stab 7/20, Stab 8/20, Stab 18/20, Stab 7/32, Stab 8/32, or Stab 18/32 (e.g., any siNA having Stab 7, 8, 11, 12, 13, 14, 15, 17, 18, 19, 20, or 32 sense or antisense strands or any combination thereof). Herein, numeric Stab chemistries can include both 2′-fluoro and 2′-OCF3 versions of the chemistries shown in Table IV. For example, “Stab 7/8” refers to both Stab 7/8 and Stab 7F/8F etc. In one embodiment, the invention features a chemically synthesized double stranded RNA molecule that directs cleavage of a PTP-1B RNA via RNA interference, wherein each strand of said RNA molecule is about 15 to about 30 nucleotides in length; one strand of the RNA molecule comprises nucleotide sequence having sufficient complementarity to the PTP-1B RNA for the RNA molecule to direct cleavage of the PTP-1B RNA via RNA interference; and wherein at least one strand of the RNA molecule optionally comprises one or more chemically modified nucleotides described herein, such as without limitation deoxynucleotides, 2′-O-methyl nucleotides, 2′-deoxy-2′-fluoro nucleotides, 2′-O-methoxyethyl nucleotides, 4′-thio nucleotides, 2′-O-trifluoromethyl nucleotides, 2′-O-ethyl-trifluoromethoxy nucleotides, 2′-O-difluoromethoxy-ethoxy nucleotides, etc. 
     In one embodiment, the invention features a medicament comprising a siNA molecule of the invention. 
     In one embodiment, the invention features an active ingredient comprising a siNA molecule of the invention. 
     In one embodiment, the invention features the use of a double-stranded short interfering nucleic acid (siNA) molecule to inhibit, down-regulate, or reduce expression of a PTP-1B gene, wherein the siNA molecule comprises one or more chemical modifications and each strand of the double-stranded siNA is independently about 15 to about 30 or more (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 or more) nucleotides long. In one embodiment, the siNA molecule of the invention is a double stranded nucleic acid molecule comprising one or more chemical modifications, where each of the two fragments of the siNA molecule independently comprise about 15 to about 40 (e.g. about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 23, 33, 34, 35, 36, 37, 38, 39, or 40) nucleotides and where one of the strands comprises at least 15 nucleotides that are complementary to nucleotide sequence of PTP-1B encoding RNA or a portion thereof. In a non-limiting example, each of the two fragments of the siNA molecule comprise about 21 nucleotides. In another embodiment, the siNA molecule is a double stranded nucleic acid molecule comprising one or more chemical modifications, where each strand is about 21 nucleotide long and where about 19 nucleotides of each fragment of the siNA molecule are base-paired to the complementary nucleotides of the other fragment of the siNA molecule, wherein at least two 3′ terminal nucleotides of each fragment of the siNA molecule are not base-paired to the nucleotides of the other fragment of the siNA molecule. In another embodiment, the siNA molecule is a double stranded nucleic acid molecule comprising one or more chemical modifications, where each strand is about 19 nucleotide long and where the nucleotides of each fragment of the siNA molecule are base-paired to the complementary nucleotides of the other fragment of the siNA molecule to form at least about 15 (e.g., 15, 16, 17, 18, or 19) base pairs, wherein one or both ends of the siNA molecule are blunt ends. In one embodiment, each of the two 3′ terminal nucleotides of each fragment of the siNA molecule is a 2′-deoxy-pyrimidine nucleotide, such as a 2′-deoxy-thymidine. In another embodiment, all nucleotides of each fragment of the siNA molecule are base-paired to the complementary nucleotides of the other fragment of the siNA molecule. In another embodiment, the siNA molecule is a double stranded nucleic acid molecule of about 19 to about 25 base pairs having a sense region and an antisense region and comprising one or more chemical modifications, where about 19 nucleotides of the antisense region are base-paired to the nucleotide sequence or a portion thereof of the RNA encoded by the PTP-1B gene. In another embodiment, about 21 nucleotides of the antisense region are base-paired to the nucleotide sequence or a portion thereof of the RNA encoded by the PTP-1B gene. In any of the above embodiments, the 5′-end of the fragment comprising said antisense region can optionally include a phosphate group. 
     In one embodiment, the invention features the use of a double-stranded short interfering nucleic acid (siNA) molecule that inhibits, down-regulates, or reduces expression of a PTP-1B gene, wherein one of the strands of the double-stranded siNA molecule is an antisense strand which comprises nucleotide sequence that is complementary to nucleotide sequence of PTP-1B RNA or a portion thereof, the other strand is a sense strand which comprises nucleotide sequence that is complementary to a nucleotide sequence of the antisense strand and wherein a majority of the pyrimidine nucleotides present in the double-stranded siNA molecule comprises a sugar modification. 
     In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that inhibits, down-regulates, or reduces expression of a PTP-1B gene, wherein one of the strands of the double-stranded siNA molecule is an antisense strand which comprises nucleotide sequence that is complementary to nucleotide sequence of PTP-1 B RNA or a portion thereof, wherein the other strand is a sense strand which comprises nucleotide sequence that is complementary to a nucleotide sequence of the antisense strand and wherein a majority of the pyrimidine nucleotides present in the double-stranded siNA molecule comprises a sugar modification. 
     In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that inhibits, down-regulates, or reduces expression of a PTP-1B gene, wherein one of the strands of the double-stranded siNA molecule is an antisense strand which comprises nucleotide sequence that is complementary to nucleotide sequence of PTP-1B RNA that encodes a protein or portion thereof, the other strand is a sense strand which comprises nucleotide sequence that is complementary to a nucleotide sequence of the antisense strand and wherein a majority of the pyrimidine nucleotides present in the double-stranded siNA molecule comprises a sugar modification. In one embodiment, each strand of the siNA molecule comprises about 15 to about 30 or more (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more) nucleotides, wherein each strand comprises at least about 15 nucleotides that are complementary to the nucleotides of the other strand. In one embodiment, the siNA molecule is assembled from two oligonucleotide fragments, wherein one fragment comprises the nucleotide sequence of the antisense strand of the siNA molecule and a second fragment comprises nucleotide sequence of the sense region of the siNA molecule. In one embodiment, the sense strand is connected to the antisense strand via a linker molecule, such as a polynucleotide linker or a non-nucleotide linker. In a further embodiment, the pyrimidine nucleotides present in the sense strand are 2′-deoxy-2′fluoro pyrimidine nucleotides and the purine nucleotides present in the sense region are 2′-deoxy purine nucleotides. In another embodiment, the pyrimidine nucleotides present in the sense strand are 2′-deoxy-2′fluoro pyrimidine nucleotides and the purine nucleotides present in the sense region are 2′-O-methyl purine nucleotides. In still another embodiment, the pyrimidine nucleotides present in the antisense strand are 2′-deoxy-2′-fluoro pyrimidine nucleotides and any purine nucleotides present in the antisense strand are 2′-deoxy purine nucleotides. In another embodiment, the antisense strand comprises one or more 2′-deoxy-2′-fluoro pyrimidine nucleotides and one or more 2′-O-methyl purine nucleotides. In another embodiment, the pyrimidine nucleotides present in the antisense strand are 2′-deoxy-2′-fluoro pyrimidine nucleotides and any purine nucleotides present in the antisense strand are 2′-O-methyl purine nucleotides. In a further embodiment the sense strand comprises a 3′-end and a 5′-end, wherein a terminal cap moiety (e.g., an inverted deoxy abasic moiety or inverted deoxy nucleotide moiety such as inverted thymidine) is present at the 5′-end, the 3′-end, or both of the 5′ and 3′ ends of the sense strand. In another embodiment, the antisense strand comprises a phosphorothioate internucleotide linkage at the 3′ end of the antisense strand. In another embodiment, the antisense strand comprises a glyceryl modification at the 3′ end. In another embodiment, the 5′-end of the antisense strand optionally includes a phosphate group. 
     In any of the above-described embodiments of a double-stranded short interfering nucleic acid (siNA) molecule that inhibits expression of a PTP-1B gene, wherein a majority of the pyrimidine nucleotides present in the double-stranded siNA molecule comprises a sugar modification, each of the two strands of the siNA molecule can comprise about 15 to about 30 or more (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more) nucleotides. In one embodiment, about 15 to about 30 or more (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more) nucleotides of each strand of the siNA molecule are base-paired to the complementary nucleotides of the other strand of the siNA molecule. In another embodiment, about 15 to about 30 or more (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more) nucleotides of each strand of the siNA molecule are base-paired to the complementary nucleotides of the other strand of the siNA molecule, wherein at least two 3′ terminal nucleotides of each strand of the siNA molecule are not base-paired to the nucleotides of the other strand of the siNA molecule. In another embodiment, each of the two 3′ terminal nucleotides of each fragment of the siNA molecule is a 2′-deoxy-pyrimidine, such as 2′-deoxy-thymidine. In one embodiment, each strand of the siNA molecule is base-paired to the complementary nucleotides of the other strand of the siNA molecule. In one embodiment, about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides of the antisense strand are base-paired to the nucleotide sequence of the PTP-1B RNA or a portion thereof. In one embodiment, about 18 to about 25 (e.g., about 18, 19, 20, 21, 22, 23, 24, or 25) nucleotides of the antisense strand are base-paired to the nucleotide sequence of the PTP-1B RNA or a portion thereof. 
     In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that inhibits expression of a PTP-1B gene, wherein one of the strands of the double-stranded siNA molecule is an antisense strand which comprises nucleotide sequence that is complementary to nucleotide sequence of PTP-1B RNA or a portion thereof, the other strand is a sense strand which comprises nucleotide sequence that is complementary to a nucleotide sequence of the antisense strand and wherein a majority of the pyrimidine nucleotides present in the double-stranded siNA molecule comprises a sugar modification, and wherein the 5′-end of the antisense strand optionally includes a phosphate group. 
     In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that inhibits expression of a PTP-1B gene, wherein one of the strands of the double-stranded siNA molecule is an anti sense strand which comprises nucleotide sequence that is complementary to nucleotide sequence of PTP-1B RNA or a portion thereof, the other strand is a sense strand which comprises nucleotide sequence that is complementary to a nucleotide sequence of the antisense strand and wherein a majority of the pyrimidine nucleotides present in the double-stranded siNA molecule comprises a sugar modification, and wherein the nucleotide sequence or a portion thereof of the anti sense strand is complementary to a nucleotide sequence of the untranslated region or a portion thereof of the PTP-1B RNA. 
     In one embodiment, the invention features a double-stranded short interfering nucleic acid (siNA) molecule that inhibits expression of a PTP-1B gene, wherein one of the strands of the double-stranded siNA molecule is an antisense strand which comprises nucleotide sequence that is complementary to nucleotide sequence of PTP-1B RNA or a portion thereof, wherein the other strand is a sense strand which comprises nucleotide sequence that is complementary to a nucleotide sequence of the antisense strand, wherein a majority of the pyrimidine nucleotides present in the double-stranded siNA molecule comprises a sugar modification, and wherein the nucleotide sequence of the antisense strand is complementary to a nucleotide sequence of the PTP-1B RNA or a portion thereof that is present in the PTP-1B RNA. 
     In one embodiment, the invention features a composition comprising a siNA molecule of the invention in a pharmaceutically acceptable carrier or diluent. 
     In a non-limiting example, the introduction of chemically-modified nucleotides into nucleic acid molecules provides a powerful tool in overcoming potential limitations of in vivo stability and bioavailability inherent to native RNA molecules that are delivered exogenously. For example, the use of chemically-modified nucleic acid molecules can enable a lower dose of a particular nucleic acid molecule for a given therapeutic effect since chemically-modified nucleic acid molecules tend to have a longer half-life in serum. Furthermore, certain chemical modifications can improve the bioavailability of nucleic acid molecules by targeting particular cells or tissues and/or improving cellular uptake of the nucleic acid molecule. Therefore, even if the activity of a chemically-modified nucleic acid molecule is reduced as compared to a native nucleic acid molecule, for example, when compared to an all-RNA nucleic acid molecule, the overall activity of the modified nucleic acid molecule can be greater than that of the native molecule due to improved stability and/or delivery of the molecule. Unlike native unmodified siNA, chemically-modified siNA can also minimize the possibility of activating interferon activity in humans. 
     In any of the embodiments of siNA molecules described herein, the antisense region of a siNA molecule of the invention can comprise a phosphorothioate internucleotide linkage at the 3′-end of said antisense region. In any of the embodiments of siNA molecules described herein, the antisense region can comprise about one to about five phosphorothioate internucleotide linkages at the 5′-end of said antisense region. In any of the embodiments of siNA molecules described herein, the 3′-terminal nucleotide overhangs of a siNA molecule of the invention can comprise ribonucleotides or deoxyribonucleotides that are chemically-modified at a nucleic acid sugar, base, or backbone. In any of the embodiments of siNA molecules described herein, the 3′-terminal nucleotide overhangs can comprise one or more universal base ribonucleotides. In any of the embodiments of siNA molecules described herein, the 3′-terminal nucleotide overhangs can comprise one or more acyclic nucleotides. 
     One embodiment of the invention provides an expression vector comprising a nucleic acid sequence encoding at least one siNA molecule of the invention in a manner that allows expression of the nucleic acid molecule. Another embodiment of the invention provides a mammalian cell comprising such an expression vector. The mammalian cell can be a human cell. The siNA molecule of the expression vector can comprise a sense region and an antisense region. The antisense region can comprise sequence complementary to a RNA or DNA sequence encoding PTP-1B and the sense region can comprise sequence complementary to the antisense region. The siNA molecule can comprise two distinct strands having complementary sense and antisense regions. The siNA molecule can comprise a single strand having complementary sense and antisense regions. 
     In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule capable of mediating RNA interference (RNAi) against PTP-1B inside a cell or reconstituted in vitro system, wherein the chemical modification comprises one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) nucleotides comprising a backbone modified internucleotide linkage having Formula I: 
     
       
         
         
             
             
         
       
     
     wherein each R1 and R2 is independently any nucleotide, non-nucleotide, or polynucleotide which can be naturally-occurring or chemically-modified, each X and Y is independently O, S, N, alkyl, or substituted alkyl, each Z and W is independently O, S, N, alkyl, substituted alkyl, O-alkyl, S-alkyl, alkaryl, aralkyl, or acetyl and wherein W, X, Y, and Z are optionally not all O. In another embodiment, a backbone modification of the invention comprises a phosphonoacetate and/or thiophosphonoacetate internucleotide linkage (see for example Sheehan et al., 2003, Nucleic Acids Research, 31, 4109-4118). 
     The chemically-modified internucleotide linkages having Formula I, for example, wherein any Z, W, X, and/or Y independently comprises a sulphur atom, can be present in one or both oligonucleotide strands of the siNA duplex, for example, in the sense strand, the antisense strand, or both strands. The siNA molecules of the invention can comprise one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) chemically-modified internucleotide linkages having Formula I at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of the sense strand, the antisense strand, or both strands. For example, an exemplary siNA molecule of the invention can comprise about 1 to about 5 or more (e.g., about 1, 2, 3, 4, 5, or more) chemically-modified internucleotide linkages having Formula I at the 5′-end of the sense strand, the antisense strand, or both strands. In another non-limiting example, an exemplary siNA molecule of the invention can comprise one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) pyrimidine nucleotides with chemically-modified internucleotide linkages having Formula 1 in the sense strand, the antisense strand, or both strands. In yet another non-limiting example, an exemplary siNA molecule of the invention can comprise one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) purine nucleotides with chemically-modified internucleotide linkages having Formula I in the sense strand, the antisense strand, or both strands. In another embodiment, a siNA molecule of the invention having internucleotide linkage(s) of Formula I also comprises a chemically-modified nucleotide or non-nucleotide having any of Formulae I-VII. 
     In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule capable of mediating RNA interference (RNAi) against PTP-1B inside a cell or reconstituted in vitro system, wherein the chemical modification comprises one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) nucleotides or non-nucleotides having Formula II: 
                         
wherein each R3, R4, R5, R6, R7, R8, R10, R11 and R12 is independently H, OH, alkyl, substituted alkyl, alkaryl or aralkyl, F, Cl, Br, CN, CF3, OCF3, OCN, O-alkyl, S-alkyl, N-alkyl, O-alkenyl, S-alkenyl, N-alkenyl, SO-alkyl, alkyl-OSH, alkyl-OH, O-alkyl-OH, O-alkyl-SH, S-alkyl-OH, S-alkyl-SH, alkyl-S-alkyl, alkyl-O-alkyl, ONO2, NO2, N3, NH2, aminoalkyl, aminoacid, aminoacyl, ONH2, O-aminoalkyl, O-aminoacid, O-aminoacyl, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalklyl amino, substituted silyl, or group having Formula I or II; R9 is O, S, CH2, S═O, CHF, or CF2, and B is a nucleosidic base such as adenine, guanine, uracil, cytosine, thymine, 2-aminoadenosine, 5-methylcytosine, 2,6-diaminopurine, or any other non-naturally occurring base that can be complementary or non-complementary to target RNA or a non-nucleosidic base such as phenyl, naphthyl, 3-nitropyrrole, 5-nitroindole, nebularine, pyridone, pyridinone, or any other non-naturally occurring universal base that can be complementary or non-complementary to target RNA. In one embodiment, R3 and/or R7 comprises a conjugate moiety and a linker (e.g., a nucleotide or non-nucleotide linker as described herein or otherwise known in the art). Non-limiting examples of conjugate moieties include ligands for cellular receptors, such as peptides derived from naturally occurring protein ligands; protein localization sequences, including cellular ZIP code sequences; antibodies; nucleic acid aptamers; vitamins and other co-factors, such as folate and N-acetylgalactosamine; polymers, such as polyethyleneglycol (PEG); phospholipids; cholesterol; steroids, and polyamines, such as PEI, spermine or spermidine.
 
     The chemically-modified nucleotide or non-nucleotide of Formula II can be present in one or both oligonucleotide strands of the siNA duplex, for example in the sense strand, the antisense strand, or both strands. The siNA molecules of the invention can comprise one or more chemically-modified nucleotides or non-nucleotides of Formula II at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of the sense strand, the antisense strand, or both strands. For example, an exemplary siNA molecule of the invention can comprise about 1 to about 5 or more (e.g., about 1, 2, 3, 4, 5, or more) chemically-modified nucleotides or non-nucleotides of Formula 11 at the 5′-end of the sense strand, the antisense strand, or both strands. In anther non-limiting example, an exemplary siNA molecule of the invention can comprise about 1 to about 5 or more (e.g., about 1, 2, 3, 4, 5, or more) chemically-modified nucleotides or non-nucleotides of Formula II at the 3′-end of the sense strand, the antisense strand, or both strands. 
     In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule capable of mediating RNA interference (RNAi) against PTP-1B inside a cell or reconstituted in vitro system, wherein the chemical modification comprises one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) nucleotides or non-nucleotides having Formula III: 
                         
wherein each R3, R4, R5, R6, R7, R8, R10, R11 and R12 is independently H, OH, alkyl, substituted alkyl, alkaryl or aralkyl, F, Cl, Br, CN, CF3, OCF3, OCN, O-alkyl, S-alkyl, N-alkyl, O-alkenyl, S-alkenyl, N-alkenyl, SO-alkyl, alkyl-OSH, alkyl-OH, O-alkyl-OH, O-alkyl-SH, S-alkyl-OH, S-alkyl-SH, alkyl-S-alkyl, alkyl-O-alkyl, ONO2, NO2, N3, NH2, aminoalkyl, aminoacid, aminoacyl, ONH2, O-aminoalkyl, O-aminoacid, O-aminoacyl, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalklylamino, substituted silyl, or group having Formula I or II; R9 is O, S, CH2, S═O, CHF, or CF2, and B is a nucleosidic base such as adenine, guanine, uracil, cytosine, thymine, 2-aminoadenosine, 5-methylcytosine, 2,6-diaminopurine, or any other non-naturally occurring base that can be employed to be complementary or non-complementary to target RNA or a non-nucleosidic base such as phenyl, naphthyl, 3-nitropyrrole, 5-nitroindole, nebularine, pyridone, pyridinone, or any other non-naturally occurring universal base that can be complementary or non-complementary to target RNA. In one embodiment, R3 and/or R7 comprises a conjugate moiety and a linker (e.g., a nucleotide or non-nucleotide linker as described herein or otherwise known in the art). Non-limiting examples of conjugate moieties include ligands for cellular receptors, such as peptides derived from naturally occurring protein ligands; protein localization sequences, including cellular ZIP code sequences; antibodies; nucleic acid aptamers; vitamins and other co-factors, such as folate and N-acetylgalactosamine; polymers, such as polyethyleneglycol (PEG); phospholipids; cholesterol; steroids, and polyamines, such as PEI, spermine or spermidine.
 
     The chemically-modified nucleotide or non-nucleotide of Formula III can be present in one or both oligonucleotide strands of the siNA duplex, for example, in the sense strand, the antisense strand, or both strands. The siNA molecules of the invention can comprise one or more chemically-modified nucleotides or non-nucleotides of Formula III at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of the sense strand, the antisense strand, or both strands. For example, an exemplary siNA molecule of the invention can comprise about 1 to about 5 or more (e.g., about 1, 2, 3, 4, 5, or more) chemically-modified nucleotide(s) or non-nucleotide(s) of Formula III at the 5′-end of the sense strand, the antisense strand, or both strands. In anther non-limiting example, an exemplary siNA molecule of the invention can comprise about 1 to about 5 or more (e.g., about 1, 2, 3, 4, 5, or more) chemically-modified nucleotide or non-nucleotide of Formula III at the 3′-end of the sense strand, the antisense strand, or both strands. 
     In another embodiment, a siNA molecule of the invention comprises a nucleotide having Formula II or III, wherein the nucleotide having Formula II or III is in an inverted configuration. For example, the nucleotide having Formula II or III is connected to the siNA construct in a 3′-3′, 3′-2′, 2′-3′, or 5′-5′ configuration, such as at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of one or both siNA strands. 
     In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule capable of mediating RNA interference (RNAi) against PTP-1B inside a cell or reconstituted in vitro system, wherein the chemical modification comprises a 5′-terminal phosphate group having Formula IV: 
                         
wherein each X and Y is independently O, S, N, alkyl, substituted alkyl, or alkylhalo; wherein each Z and W is independently O, S, N, alkyl, substituted alkyl, O-alkyl, S-alkyl, alkaryl, aralkyl, alkylhalo, or acetyl; and wherein W, X, Y and Z are not all O.
 
     In one embodiment, the invention features a siNA molecule having a 5′-terminal phosphate group having Formula IV on the target-complementary strand, for example, a strand complementary to a target RNA, wherein the siNA molecule comprises an all RNA siNA molecule. In another embodiment, the invention features a siNA molecule having a 5′-terminal phosphate group having Formula IV on the target-complementary strand wherein the siNA molecule also comprises about 1 to about 3 (e.g., about 1, 2, or 3) nucleotide 3′-terminal nucleotide overhangs having about 1 to about 4 (e.g., about 1, 2, 3, or 4) deoxyribonucleotides on the 3′-end of one or both strands. In another embodiment, a 5′-terminal phosphate group having Formula IV is present on the target-complementary strand of a siNA molecule of the invention, for example a siNA molecule having chemical modifications having any of Formulae I-VII. 
     In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule capable of mediating RNA interference (RNAi) against PTP-1B inside a cell or reconstituted in vitro system, wherein the chemical modification comprises one or more phosphorothioate internucleotide linkages. For example, in a non-limiting example, the invention features a chemically-modified short interfering nucleic acid (siNA) having about 1, 2, 3, 4, 5, 6, 7, 8 or more phosphorothioate internucleotide linkages in one siNA strand. In yet another embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) individually having about 1, 2, 3, 4, 5, 6, 7, 8 or more phosphorothioate internucleotide linkages in both siNA strands. The phosphorothioate internucleotide linkages can be present in one or both oligonucleotide strands of the siNA duplex, for example in the sense strand, the antisense strand, or both strands. The siNA molecules of the invention can comprise one or more phosphorothioate internucleotide linkages at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the sense strand, the antisense strand, or both strands. For example, an exemplary siNA molecule of the invention can comprise about 1 to about 5 or more (e.g., about 1, 2, 3, 4, 5, or more) consecutive phosphorothioate internucleotide linkages at the 5′-end of the sense strand, the antisense strand, or both strands. In another non-limiting example, an exemplary siNA molecule of the invention can comprise one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) pyrimidine phosphorothioate internucleotide linkages in the sense strand, the antisense strand, or both strands. In yet another non-limiting example, an exemplary siNA molecule of the invention can comprise one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) purine phosphorothioate internucleotide linkages in the sense strand, the antisense strand, or both strands. 
     In one embodiment, the invention features a siNA molecule, wherein the sense strand comprises one or more, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy and/or about one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the sense strand; and wherein the antisense strand comprises about 1 to about 10 or more, specifically about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the antisense strand. In another embodiment, one or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more, pyrimidine nucleotides of the sense and/or antisense siNA strand are chemically-modified with 2′-deoxy, 2′-O-methyl, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or 2′-deoxy-2′-fluoro nucleotides, with or without one or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more, phosphorothioate internucleotide linkages and/or a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends, being present in the same or different strand. 
     In another embodiment, the invention features a siNA molecule, wherein the sense strand comprises about 1 to about 5, specifically about 1, 2, 3, 4, or 5 phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or one or more (e.g., about 1, 2, 3, 4, 5, or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3-end, the 5′-end, or both of the 3′- and 5′-ends of the sense strand; and wherein the antisense strand comprises about 1 to about 5 or more, specifically about 1, 2, 3, 4, 5, or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the antisense strand. In another embodiment, one or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more, pyrimidine nucleotides of the sense and/or antisense siNA strand are chemically-modified with 2′-deoxy, 2′-O-methyl, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or 2′-deoxy-2′-fluoro nucleotides, with or without about 1 to about 5 or more, for example about 1, 2, 3, 4, 5, or more phosphorothioate internucleotide linkages and/or a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends, being present in the same or different strand. 
     In one embodiment, the invention features a siNA molecule, wherein the antisense strand comprises one or more, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more phosphorothioate internucleotide linkages, and/or about one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the sense strand; and wherein the antisense strand comprises about 1 to about 10 or more, specifically about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the antisense strand. In another embodiment, one or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more pyrimidine nucleotides of the sense and/or antisense siNA strand are chemically-modified with 2′-deoxy, 2′-O-methyl, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or 2′-deoxy-2′-fluoro nucleotides, with or without one or more, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more phosphorothioate internucleotide linkages and/or a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends, being present in the same or different strand. 
     In another embodiment, the invention features a siNA molecule, wherein the antisense strand comprises about 1 to about 5 or more, specifically about 1, 2, 3, 4, 5 or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the sense strand; and wherein the antisense strand comprises about 1 to about 5 or more, specifically about 1, 2, 3, 4, 5 or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the antisense strand. In another embodiment, one or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more pyrimidine nucleotides of the sense and/or antisense siNA strand are chemically-modified with 2′-deoxy, 2′-O-methyl, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy, 4′-thio and/or 2′-deoxy-2′-fluoro nucleotides, with or without about 1 to about 5, for example about 1, 2, 3, 4, 5 or more phosphorothioate internucleotide linkages and/or a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends, being present in the same or different strand. 
     In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule having about 1 to about 5 or more (specifically about 1, 2, 3, 4, 5 or more) phosphorothioate internucleotide linkages in each strand of the siNA molecule. 
     In another embodiment, the invention features a siNA molecule comprising 2′-5′ internucleotide linkages. The 2′-5′ internucleotide linkage(s) can be at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of one or both siNA sequence strands. In addition, the 2′-5′ internucleotide linkage(s) can be present at various other positions within one or both siNA sequence strands, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more including every internucleotide linkage of a pyrimidine nucleotide in one or both strands of the siNA molecule can comprise a 2′-5′ internucleotide linkage, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more including every internucleotide linkage of a purine nucleotide in one or both strands of the siNA molecule can comprise a 2′-5′ internucleotide linkage. 
     In another embodiment, a chemically-modified siNA molecule of the invention comprises a duplex having two strands, one or both of which can be chemically-modified, wherein each strand is independently about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides in length, wherein the duplex has about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) base pairs, and wherein the chemical modification comprises a structure having any of Formulae I-VII. For example, an exemplary chemically-modified siNA molecule of the invention comprises a duplex having two strands, one or both of which can be chemically-modified with a chemical modification having any of Formulae I-VII or any combination thereof, wherein each strand consists of about 21 nucleotides, each having a 2-nucleotide 3′-terminal nucleotide overhang, and wherein the duplex has about 19 base pairs. In another embodiment, a siNA molecule of the invention comprises a single stranded hairpin structure, wherein the siNA is about 36 to about 70 (e.g., about 36, 40, 45, 50, 55, 60, 65, or 70) nucleotides in length having about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) base pairs, and wherein the siNA can include a chemical modification comprising a structure having any of Formulae I-VII or any combination thereof. For example, an exemplary chemically-modified siNA molecule of the invention comprises a linear oligonucleotide having about 42 to about 50 (e.g., about 42, 43, 44, 45, 46, 47, 48, 49, or 50) nucleotides that is chemically-modified with a chemical modification having any of Formulae I-VII or any combination thereof, wherein the linear oligonucleotide forms a hairpin structure having about 19 to about 21 (e.g., 19, 20, or 21) base pairs and a 2-nucleotide 3′-terminal nucleotide overhang. In another embodiment, a linear hairpin siNA molecule of the invention contains a stem loop motif, wherein the loop portion of the siNA molecule is biodegradable. For example, a linear hairpin siNA molecule of the invention is designed such that degradation of the loop portion of the siNA molecule in vivo can generate a double-stranded siNA molecule with 3′-terminal overhangs, such as 3′-terminal nucleotide overhangs comprising about 2 nucleotides. 
     In another embodiment, a siNA molecule of the invention comprises a hairpin structure, wherein the siNA is about 25 to about 50 (e.g., about 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50) nucleotides in length having about 3 to about 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) base pairs, and wherein the siNA can include one or more chemical modifications comprising a structure having any of Formulae I-VII or any combination thereof. For example, an exemplary chemically-modified siNA molecule of the invention comprises a linear oligonucleotide having about 25 to about 35 (e.g., about 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) nucleotides that is chemically-modified with one or more chemical modifications having any of Formulae I-VII or any combination thereof, wherein the linear oligonucleotide forms a hairpin structure having about 3 to about 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) base pairs and a 5′-terminal phosphate group that can be chemically modified as described herein (for example a 5′-terminal phosphate group having Formula IV). In another embodiment, a linear hairpin siNA molecule of the invention contains a stem loop motif, wherein the loop portion of the siNA molecule is biodegradable. In one embodiment, a linear hairpin siNA molecule of the invention comprises a loop portion comprising a non-nucleotide linker. 
     In another embodiment, a siNA molecule of the invention comprises an asymmetric hairpin structure, wherein the siNA is about 25 to about 50 (e.g., about 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50) nucleotides in length having about 3 to about 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) base pairs, and wherein the siNA can include one or more chemical modifications comprising a structure having any of Formulae I-VII or any combination thereof. For example, an exemplary chemically-modified siNA molecule of the invention comprises a linear oligonucleotide having about 25 to about 35 (e.g., about 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, or 35) nucleotides that is chemically-modified with one or more chemical modifications having any of Formulae I-VII or any combination thereof, wherein the linear oligonucleotide forms an asymmetric hairpin structure having about 3 to about 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) base pairs and a 5′-terminal phosphate group that can be chemically modified as described herein (for example a 5′-terminal phosphate group having Formula IV). In one embodiment, an asymmetric hairpin siNA molecule of the invention contains a stem loop motif, wherein the loop portion of the siNA molecule is biodegradable. In another embodiment, an asymmetric hairpin siNA molecule of the invention comprises a loop portion comprising a non-nucleotide linker. 
     In another embodiment, a siNA molecule of the invention comprises an asymmetric double stranded structure having separate polynucleotide strands comprising sense and antisense regions, wherein the antisense region is about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides in length, wherein the sense region is about 3 to about 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) nucleotides in length, wherein the sense region and the antisense region have at least 3 complementary nucleotides, and wherein the siNA can include one or more chemical modifications comprising a structure having any of Formulae I-VII or any combination thereof. For example, an exemplary chemically-modified siNA molecule of the invention comprises an asymmetric double stranded structure having separate polynucleotide strands comprising sense and antisense regions, wherein the antisense region is about 18 to about 23 (e.g., about 18, 19, 20, 21, 22, or 23) nucleotides in length and wherein the sense region is about 3 to about 15 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15) nucleotides in length, wherein the sense region the antisense region have at least 3 complementary nucleotides, and wherein the siNA can include one or more chemical modifications comprising a structure having any of Formulae I-VII or any combination thereof. In another embodiment, the asymmetric double stranded siNA molecule can also have a 5′-terminal phosphate group that can be chemically modified as described herein (for example a 5′-terminal phosphate group having Formula IV). 
     In another embodiment, a siNA molecule of the invention comprises a circular nucleic acid molecule, wherein the siNA is about 38 to about 70 (e.g., about 38, 40, 45, 50, 55, 60, 65, or 70) nucleotides in length having about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) base pairs, and wherein the siNA can include a chemical modification, which comprises a structure having any of Formulae I-VII or any combination thereof For example, an exemplary chemically-modified siNA molecule of the invention comprises a circular oligonucleotide having about 42 to about 50 (e.g., about 42, 43, 44, 45, 46, 47, 48, 49, or 50) nucleotides that is chemically-modified with a chemical modification having any of Formulae I-VII or any combination thereof, wherein the circular oligonucleotide forms a dumbbell shaped structure having about 19 base pairs and 2 loops. 
     In another embodiment, a circular siNA molecule of the invention contains two loop motifs, wherein one or both loop portions of the siNA molecule is biodegradable. For example, a circular siNA molecule of the invention is designed such that degradation of the loop portions of the siNA molecule in vivo can generate a double-stranded siNA molecule with 3′-terminal overhangs, such as 3′-terminal nucleotide overhangs comprising about 2 nucleotides. 
     In one embodiment, a siNA molecule of the invention comprises at least one (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) abasic moiety, for example a compound having Formula V: 
                         
wherein each R3, R4, R5, R6, R7, R8, R10, R11, R12, and R13 is independently H, OH, alkyl, substituted alkyl, alkaryl or aralkyl, F, Cl, Br, CN, CF3, OCF3, OCN, O-alkyl, S-alkyl, N-alkyl, O-alkenyl, S-alkenyl, N-alkenyl, SO-alkyl, alkyl-OSH, alkyl-OH, O-alkyl-OH, O-alkyl-SH, S-alkyl-OH, S-alkyl-SH, alkyl-S-alkyl, alkyl-O-alkyl, ONO2, NO2, N3, NH2, aminoalkyl, aminoacid, aminoacyl, ONH2, O-aminoalkyl, O-aminoacid, O-aminoacyl, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalklylamino, substituted silyl, or group having Formula I or II; R9 is O, S, CH2, S═O, CHF, or CF2. In one embodiment, R3 and/or R7 comprises a conjugate moiety and a linker (e.g., a nucleotide or non-nucleotide linker as described herein or otherwise known in the art). Non-limiting examples of conjugate moieties include ligands for cellular receptors, such as peptides derived from naturally occurring protein ligands; protein localization sequences, including cellular ZIP code sequences; antibodies; nucleic acid aptamers; vitamins and other co-factors, such as folate and N-acetylgalactosamine; polymers, such as polyethyleneglycol (PEG); phospholipids; cholesterol; steroids, and polyamines, such as PEI, spermine or spermidine.
 
     In one embodiment, a siNA molecule of the invention comprises at least one (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) inverted abasic moiety, for example a compound having Formula VI: 
                         
wherein each R3, R4, R5, R6, R7, R8, R10, R11, R12, and R13 is independently H, OH, alkyl, substituted alkyl, alkaryl or aralkyl, F, Cl, Br, CN, CF3, OCF3, OCN, O-alkyl, S-alkyl, N-alkyl, O-alkenyl, S-alkenyl, N-alkenyl, SO-alkyl, alkyl-OSH, alkyl-OH, O-alkyl-OH, O-alkyl-SH, S-alkyl-OH, S-alkyl-SH, alkyl-S-alkyl, alkyl-O-alkyl, ONO2, NO2, N3, NH2, aminoalkyl, aminoacid, aminoacyl, ONH2, O-aminoalkyl, O-aminoacid, O-aminoacyl, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalklylamino, substituted silyl, or group having Formula I or II; R9 is O, S, CH2, S═O, CHF, or CF2, and either R2, R3, R8 or R13 serve as points of attachment to the siNA molecule of the invention. In one embodiment, R3 and/or R7 comprises a conjugate moiety and a linker (e.g., a nucleotide or non-nucleotide linker as described herein or otherwise known in the art). Non-limiting examples of conjugate moieties include ligands for cellular receptors, such as peptides derived from naturally occurring protein ligands; protein localization sequences, including cellular ZIP code sequences; antibodies; nucleic acid aptamers; vitamins and other co-factors, such as folate and N-acetylgalactosamine; polymers, such as polyethyleneglycol (PEG); phospholipids; cholesterol; steroids, and polyamines, such as PEI, spermine or spermidine.
 
     In another embodiment, a siNA molecule of the invention comprises at least one (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) substituted polyalkyl moieties, for example a compound having Formula VII: 
                         
wherein each n is independently an integer from 1 to 12, each R1, R2 and R3 is independently H, OH, alkyl, substituted alkyl, alkaryl or aralkyl, F, Cl, Br, CN, CF3, OCF3, OCN, O-alkyl, S-alkyl, N-alkyl, O-alkenyl, S-alkenyl, N-alkenyl, SO-alkyl, alkyl-OSH, alkyl-OH, O-alkyl-OH, O-alkyl-SH, S-alkyl-OH, S-alkyl-SH, alkyl-S-alkyl, alkyl-O-alkyl, ONO2, NO2, N3, NH2, aminoalkyl, aminoacid, aminoacyl, ONH2, O-aminoalkyl, O-aminoacid, O-aminoacyl, heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalklylamino, substituted silyl, or a group having Formula I, and R1, R2 or R3 serves as points of attachment to the siNA molecule of the invention. In one embodiment, R3 and/or R1 comprises a conjugate moiety and a linker (e.g., a nucleotide or non-nucleotide linker as described herein or otherwise known in the art). Non-limiting examples of conjugate moieties include ligands for cellular receptors, such as peptides derived from naturally occurring protein ligands; protein localization sequences, including cellular ZIP code sequences; antibodies; nucleic acid aptamers; vitamins and other co-factors, such as folate and N-acetylgalactosamine; polymers, such as polyethyleneglycol (PEG); phospholipids; cholesterol; steroids, and polyamines, such as PEI, spermine or spermidine.
 
     By “ZIP code” sequences is meant, any peptide or protein sequence that is involved in cellular topogenic signaling mediated transport (see for example Ray et al., 2004,  Science,  306(1501): 1505) 
     In another embodiment, the invention features a compound having Formula VII, wherein R1 and R2 are hydroxyl (OH) groups, n=1, and R3 comprises O and is the point of attachment to the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of one or both strands of a double-stranded siNA molecule of the invention or to a single-stranded siNA molecule of the invention. This modification is referred to herein as “glyceryl” (for example modification 6 in  FIG. 10 ). 
     In another embodiment, a chemically modified nucleoside or non-nucleoside (e.g. a moiety having any of Formula V, VI or VII) of the invention is at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of a siNA molecule of the invention. For example, chemically modified nucleoside or non-nucleoside (e.g., a moiety having Formula V, VI or VII) can be present at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of the antisense strand, the sense strand, or both antisense and sense strands of the siNA molecule. In one embodiment, the chemically modified nucleoside or non-nucleoside (e.g., a moiety having Formula V, VI or VII) is present at the 5′-end and 3′-end of the sense strand and the 3′-end of the antisense strand of a double stranded siNA molecule of the invention. In one embodiment, the chemically modified nucleoside or non-nucleoside (e.g., a moiety having Formula V, VI or VII) is present at the terminal position of the 5′-end and 3′-end of the sense strand and the 3′-end of the antisense strand of a double stranded siNA molecule of the invention. In one embodiment, the chemically modified nucleoside or non-nucleoside (e.g., a moiety having Formula V, VI or VII) is present at the two terminal positions of the 5′-end and 3′-end of the sense strand and the 3′-end of the antisense strand of a double stranded siNA molecule of the invention. In one embodiment, the chemically modified nucleoside or non-nucleoside (e.g., a moiety having Formula V, VI or VII) is present at the penultimate position of the 5′-end and 3′-end of the sense strand and the 3′-end of the antisense strand of a double stranded siNA molecule of the invention. In addition, a moiety having Formula VII can be present at the 3′-end or the 5′-end of a hairpin siNA molecule as described herein. 
     In another embodiment, a siNA molecule of the invention comprises an abasic residue having Formula V or VI, wherein the abasic residue having Formula VI or VI is connected to the siNA construct in a 3′-3′, 3′-2′, 2′-3′, or 5′-5′ configuration, such as at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of one or both siNA strands. 
     In one embodiment, a siNA molecule of the invention comprises one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) locked nucleic acid (LNA) nucleotides, for example, at the 5′-end, the 3′-end, both of the 5′ and 3′-ends, or any combination thereof, of the siNA molecule. 
     In one embodiment, a siNA molecule of the invention comprises one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) 4′-thio nucleotides, for example, at the 5′-end, the 3′-end, both of the 5′ and 3′-ends, or any combination thereof, of the siNA molecule. 
     In another embodiment, a siNA molecule of the invention comprises one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) acyclic nucleotides, for example, at the 5′-end, the 3′-end, both of the 5′ and 3′-ends, or any combination thereof, of the siNA molecule. 
     In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention comprising a sense region, wherein any (e.g., one or more or all) pyrimidine nucleotides present in the sense region are 2′-deoxy-2′-fluoro pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro pyrimidine nucleotides), and wherein any (e.g., one or more or all) purine nucleotides present in the sense region are 2′-deoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-deoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-deoxy purine nucleotides). 
     In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention comprising a sense region, wherein any (e.g., one or more or all) pyrimidine nucleotides present in the sense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), and wherein any (e.g., one or more or all) purine nucleotides present in the sense region are 2′-deoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-deoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-deoxy purine nucleotides), wherein any nucleotides comprising a 3′-terminal nucleotide overhang that are present in said sense region are 2′-deoxy nucleotides. 
     In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention comprising a sense region, wherein any (e.g., one or more or all) pyrimidine nucleotides present in the sense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), and wherein any (e.g., one or more or all) purine nucleotides present in the sense region are 2′-O-methyl purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides). 
     In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention comprising a sense region, wherein any (e.g., one or more or all) pyrimidine nucleotides present in the sense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-di fluoromethoxy-ethoxy pyrimidine nucleotides), wherein any (e.g., one or more or all) purine nucleotides present in the sense region are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides), and wherein any nucleotides comprising a 3′-terminal nucleotide overhang that are present in said sense region are 2′-deoxy nucleotides. 
     In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention comprising an antisense region, wherein any (e.g., one or more or all) pyrimidine nucleotides present in the antisense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), and wherein any (e.g., one or more or all) purine nucleotides present in the antisense region are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-di fluoromethoxy-ethoxy purine nucleotides). 
     In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention comprising an antisense region, wherein any (e.g., one or more or all) pyrimidine nucleotides present in the antisense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), wherein any (e.g., one or more or all) purine nucleotides present in the antisense region are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides), and wherein any nucleotides comprising a 3′-terminal nucleotide overhang that are present in said antisense region are 2′-deoxy nucleotides. 
     In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention comprising an antisense region, wherein any (e.g., one or more or all) pyrimidine nucleotides present in the antisense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), and wherein any (e.g., one or more or all) purine nucleotides present in the antisense region are 2′-deoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-deoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-deoxy purine nucleotides). 
     In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention comprising an antisense region, wherein any (e.g., one or more or all) pyrimidine nucleotides present in the antisense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), and wherein any (e.g., one or more or all) purine nucleotides present in the anti sense region are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides). 
     In one embodiment, the invention features a chemically-modified short interfering nucleic acid (siNA) molecule of the invention capable of mediating RNA interference (RNAi) against PTP-1B inside a cell or reconstituted in vitro system comprising a sense region, wherein one or more pyrimidine nucleotides present in the sense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), and one or more purine nucleotides present in the sense region are 2′-deoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-deoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-deoxy purine nucleotides), and an antisense region, wherein one or more pyrimidine nucleotides present in the antisense region are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), and one or more purine nucleotides present in the antisense region are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides). The sense region and/or the antisense region can have a terminal cap modification, such as any modification described herein or shown in  FIG. 10 , that is optionally present at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of the sense and/or antisense sequence. The sense and/or antisense region can optionally further comprise a 3′-terminal nucleotide overhang having about 1 to about 4 (e.g., about 1, 2, 3, or 4) 2′-deoxynucleotides. The overhang nucleotides can further comprise one or more (e.g., about 1, 2, 3, 4 or more) phosphorothioate, phosphonoacetate, and/or thiophosphonoacetate internucleotide linkages. Non-limiting examples of these chemically-modified siNAs are shown in  FIGS. 4 and 5  and Tables III and IV herein. In any of these described embodiments, the purine nucleotides present in the sense region are alternatively 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides) and one or more purine nucleotides present in the antisense region are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides). Also, in any of these embodiments, one or more purine nucleotides present in the sense region are alternatively purine ribonucleotides (e.g., wherein all purine nucleotides are purine ribonucleotides or alternately a plurality of purine nucleotides are purine ribonucleotides) and any purine nucleotides present in the antisense region are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides). Additionally, in any of these embodiments, one or more purine nucleotides present in the sense region and/or present in the antisense region are alternatively selected from the group consisting of 2′-deoxy nucleotides, locked nucleic acid (LNA) nucleotides, 2′-methoxyethyl nucleotides, 4′-thionucleotides, 2′-O-trifluoromethyl nucleotides, 2′-O-ethyl-trifluoromethoxy nucleotides, 2′-O-difluoromethoxy-ethoxy nucleotides and 2′-O-methyl nucleotides (e.g., wherein all purine nucleotides are selected from the group consisting of 2′-deoxy nucleotides, locked nucleic acid (LNA) nucleotides, 2′-methoxyethyl nucleotides, 4′-thionucleotides, 2′-O-trifluoromethyl nucleotides, 2′-O-ethyl-trifluoromethoxy nucleotides, 2′-O-difluoromethoxy-ethoxy nucleotides and 2′-O-methyl nucleotides or alternately a plurality of purine nucleotides are selected from the group consisting of 2′-deoxy nucleotides, locked nucleic acid (LNA) nucleotides, 2′-methoxyethyl nucleotides, 4′-thionucleotides, 2′-O-trifluoromethyl nucleotides, 2′-O-ethyl-trifluoromethoxy nucleotides, 2′-O-difluoromethoxy-ethoxy nucleotides and 2′-O-methyl nucleotides). 
     In another embodiment, any modified nucleotides present in the siNA molecules of the invention, preferably in the antisense strand of the siNA molecules of the invention, but also optionally in the sense and/or both antisense and sense strands, comprise modified nucleotides having properties or characteristics similar to naturally occurring ribonucleotides. For example, the invention features siNA molecules including modified nucleotides having a Northern conformation (e.g., Northern pseudorotation cycle, see for example Saenger,  Principles of Nucleic Acid Structure , Springer-Verlag ed., 1984). As such, chemically modified nucleotides present in the siNA molecules of the invention, preferably in the antisense strand of the siNA molecules of the invention, but also optionally in the sense and/or both antisense and sense strands, are resistant to nuclease degradation while at the same time maintaining the capacity to mediate RNAi. Non-limiting examples of nucleotides having a northern configuration include locked nucleic acid (LNA) nucleotides (e.g., 2′-O, 4′-C-methylene-(D-ribofuranosyl) nucleotides); 2′-methoxyethoxy (MOE) nucleotides; 2′-methyl-thio-ethyl, 2′-deoxy-2′-fluoro nucleotides, 2′-deoxy-2′-chloro nucleotides, 2′-azido nucleotides, 2′-O-trifluoromethyl nucleotides, 2′-O-ethyl-trifluoromethoxy nucleotides, 2′-O-difluoromethoxy-ethoxy nucleotides, 4′-thio nucleotides and 2′-O-methyl nucleotides. 
     In one embodiment, the sense strand of a double stranded siNA molecule of the invention comprises a terminal cap moiety, (see for example  FIG. 10 ) such as an inverted deoxyabasic moiety, at the 3′-end, 5′-end, or both 3′ and 5′-ends of the sense strand. 
     In one embodiment, the invention features a chemically-modified short interfering nucleic acid molecule (siNA) capable of mediating RNA interference (RNAi) against PTP-1B inside a cell or reconstituted in vitro system, wherein the chemical modification comprises a conjugate covalently attached to the chemically-modified siNA molecule. Non-limiting examples of conjugates contemplated by the invention include conjugates and ligands described in Vargeese et al., U.S. Ser. No. 10/427,160, filed Apr. 30, 2003, incorporated by reference herein in its entirety, including the drawings. In another embodiment, the conjugate is covalently attached to the chemically-modified siNA molecule via a biodegradable linker. In one embodiment, the conjugate molecule is attached at the 3′-end of either the sense strand, the antisense strand, or both strands of the chemically-modified siNA molecule. In another embodiment, the conjugate molecule is attached at the 5′-end of either the sense strand, the antisense strand, or both strands of the chemically-modified siNA molecule. In yet another embodiment, the conjugate molecule is attached both the 3′-end and 5′-end of either the sense strand, the antisense strand, or both strands of the chemically-modified siNA molecule, or any combination thereof. In one embodiment, a conjugate molecule of the invention comprises a molecule that facilitates delivery of a chemically-modified siNA molecule into a biological system, such as a cell. In another embodiment, the conjugate molecule attached to the chemically-modified siNA molecule is a ligand for a cellular receptor, such as peptides derived from naturally occurring protein ligands; protein localization sequences, including cellular ZIP code sequences; antibodies; nucleic acid aptamers; vitamins and other co-factors, such as folate and N-acetylgalactosamine; polymers, such as polyethyleneglycol (PEG); phospholipids; cholesterol; steroids, and polyamines, such as PEI, spermine or spermidine. Examples of specific conjugate molecules contemplated by the instant invention that can be attached to chemically-modified siNA molecules are described in Vargeese et al., U.S. Ser. No. 10/201,394, filed Jul. 22, 2002 incorporated by reference herein. The type of conjugates used and the extent of conjugation of siNA molecules of the invention can be evaluated for improved pharmacokinetic profiles, bioavailability, and/or stability of siNA constructs while at the same time maintaining the ability of the siNA to mediate RNAi activity. As such, one skilled in the art can screen siNA constructs that are modified with various conjugates to determine whether the siNA conjugate complex possesses improved properties while maintaining the ability to mediate RNAi, for example in animal models as are generally known in the art. 
     In one embodiment, the invention features a short interfering nucleic acid (siNA) molecule of the invention, wherein the siNA further comprises a nucleotide, non-nucleotide, or mixed nucleotide/non-nucleotide linker that joins the sense region of the siNA to the antisense region of the siNA. In one embodiment, a nucleotide, non-nucleotide, or mixed nucleotide/non-nucleotide linker is used, for example, to attach a conjugate moiety to the siNA. In one embodiment, a nucleotide linker of the invention can be a linker of ≧2 nucleotides in length, for example about 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides in length. In another embodiment, the nucleotide linker can be a nucleic acid aptamer. By “aptamer” or “nucleic acid aptamer” as used herein is meant a nucleic acid molecule that binds specifically to a target molecule wherein the nucleic acid molecule has sequence that comprises a sequence recognized by the target molecule in its natural setting. Alternately, an aptamer can be a nucleic acid molecule that binds to a target molecule where the target molecule does not naturally bind to a nucleic acid. The target molecule can be any molecule of interest. For example, the aptamer can be used to bind to a ligand-binding domain of a protein, thereby preventing interaction of the naturally occurring ligand with the protein. This is a non-limiting example and those in the art will recognize that other embodiments can be readily generated using techniques generally known in the art. (See, for example, Gold et al., 1995,  Annu. Rev. Biochem.,  64, 763; Brody and Gold, 2000,  J. Biotechnol.,  74, 5; Sun, 2000,  Curr. Opin. Mol. Ther.,  2, 100; Kusser, 2000,  J. Biotechnol.,  74, 27; Hermann and Patel, 2000,  Science,  287, 820; and Jayasena, 1999,  Clinical Chemistry,  45, 1628.) 
     In yet another embodiment, a non-nucleotide linker of the invention comprises abasic nucleotide, polyether, polyamine, polyamide, peptide, carbohydrate, lipid, polyhydrocarbon, or other polymeric compounds (e.g. polyethylene glycols such as those having between 2 and 100 ethylene glycol units). Specific examples include those described by Seela and Kaiser,  Nucleic Acids Res.  1990, 18:6353 and  Nucleic Acids Res.  1987, 15:3113; Cload and Schepartz,  J. Am. Chem. Soc.  1991, 113:6324; Richardson and Schepartz,  J. Am. Chem. Soc.  1991, 113:5109; Ma et al.,  Nucleic Acids Res.  1993, 21:2585 and  Biochemistry  1993, 32:1751; Durand et al.,  Nucleic Acids Res.  1990, 18:6353; McCurdy et al.,  Nucleosides  &amp;  Nucleotides  1991, 10:287; Jschke et al.,  Tetrahedron Lett.  1993, 34:301; Ono et al.,  Biochemistry  1991, 30:9914; Arnold et al., International Publication No. WO 89/02439; Usman et al., International Publication No. WO 95/06731; Dudycz et al., International Publication No. WO 95/11910 and Ferentz and Verdine,  J. Am. Chem. Soc.  1991, 113:4000, all hereby incorporated by reference herein. A “non-nucleotide” further means any group or compound that can be incorporated into a nucleic acid chain in the place of one or more nucleotide units, including either sugar and/or phosphate substitutions, and allows the remaining bases to exhibit their enzymatic activity. The group or compound can be abasic in that it does not contain a commonly recognized nucleotide base, such as adenosine, guanine, cytosine, uracil or thymine, for example at the C1 position of the sugar. 
     In one embodiment, the invention features a short interfering nucleic acid (siNA) molecule capable of mediating RNA interference (RNAi) inside a cell or reconstituted in vitro system, wherein one or both strands of the siNA molecule that are assembled from two separate oligonucleotides do not comprise any ribonucleotides. For example, a siNA molecule can be assembled from a single oligonculeotide where the sense and antisense regions of the siNA comprise separate oligonucleotides that do not have any ribonucleotides (e.g., nucleotides having a 2′-OH group) present in the oligonucleotides. In another example, a siNA molecule can be assembled from a single oligonculeotide where the sense and antisense regions of the siNA are linked or circularized by a nucleotide or non-nucleotide linker as described herein, wherein the oligonucleotide does not have any ribonucleotides (e.g., nucleotides having a 2′-OH group) present in the oligonucleotide. Applicant has surprisingly found that the presence of ribonucleotides (e.g., nucleotides having a 2′-hydroxyl group) within the siNA molecule is not required or essential to support RNAi activity. As such, in one embodiment, all positions within the siNA can include chemically modified nucleotides and/or non-nucleotides such as nucleotides and or non-nucleotides having Formula I, II, III, IV, V, VI, or VII or any combination thereof to the extent that the ability of the siNA molecule to support RNAi activity in a cell is maintained. 
     In one embodiment, a siNA molecule of the invention is a single stranded siNA molecule that mediates RNAi activity in a cell or reconstituted in vitro system comprising a single stranded polynucleotide having complementarity to a target nucleic acid sequence. In another embodiment, the single stranded siNA molecule of the invention comprises a 5′-terminal phosphate group. In another embodiment, the single stranded siNA molecule of the invention comprises a 5′-terminal phosphate group and a 3′-terminal phosphate group (e.g., a 2′,3′-cyclic phosphate). In another embodiment, the single stranded siNA molecule of the invention comprises about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides. In yet another embodiment, the single stranded siNA molecule of the invention comprises one or more chemically modified nucleotides or non-nucleotides described herein. For example, all the positions within the siNA molecule can include chemically-modified nucleotides such as nucleotides having any of Formulae I-VII, or any combination thereof to the extent that the ability of the siNA molecule to support RNAi activity in a cell is maintained. 
     In one embodiment, a siNA molecule of the invention is a single stranded siNA molecule that mediates RNAi activity in a cell or reconstituted in vitro system comprising a single stranded polynucleotide having complementarity to a target nucleic acid sequence, wherein one or more pyrimidine nucleotides present in the siNA are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy pyrimidine nucleotides), and wherein any purine nucleotides present in the antisense region are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-O-methyl, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, or 2′-O-difluoromethoxy-ethoxy purine nucleotides), and a terminal cap modification, such as any modification described herein or shown in  FIG. 10 , that is optionally present at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of the antisense sequence. The siNA optionally further comprises about 1 to about 4 or more (e.g., about 1, 2, 3, 4 or more) terminal 2′-deoxynucleotides at the 3′-end of the siNA molecule, wherein the terminal nucleotides can further comprise one or more (e.g., 1, 2, 3, 4 or more) phosphorothioate, phosphonoacetate, and/or thiophosphonoacetate internucleotide linkages, and wherein the siNA optionally further comprises a terminal phosphate group, such as a 5′-terminal phosphate group. In any of these embodiments, any purine nucleotides present in the antisense region are alternatively 2′-deoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-deoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-deoxy purine nucleotides). Also, in any of these embodiments, any purine nucleotides present in the siNA (i.e., purine nucleotides present in the sense and/or antisense region) can alternatively be locked nucleic acid (LNA) nucleotides (e.g., wherein all purine nucleotides are LNA nucleotides or alternately a plurality of purine nucleotides are LNA nucleotides). Also, in any of these embodiments, any purine nucleotides present in the siNA are alternatively 2′-methoxyethyl purine nucleotides (e.g., wherein all purine nucleotides are 2′-methoxyethyl purine nucleotides or alternately a plurality of purine nucleotides are 2′-methoxyethyl purine nucleotides). In another embodiment, any modified nucleotides present in the single stranded siNA molecules of the invention comprise modified nucleotides having properties or characteristics similar to naturally occurring ribonucleotides. For example, the invention features siNA molecules including modified nucleotides having a Northern conformation (e.g., Northern pseudorotation cycle, see for example Saenger,  Principles of Nucleic Acid Structure , Springer-Verlag ed., 1984). As such, chemically modified nucleotides present in the single stranded siNA molecules of the invention are preferably resistant to nuclease degradation while at the same time maintaining the capacity to mediate RNAi. 
     In one embodiment, a siNA molecule of the invention comprises chemically modified nucleotides or non-nucleotides (e.g., having any of Formulae I-VII, such as 2′-deoxy, 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy or 2′-O-methyl nucleotides) at alternating positions within one or more strands or regions of the siNA molecule. For example, such chemical modifications can be introduced at every other position of a RNA based siNA molecule, starting at either the first or second nucleotide from the 3′-end or 5′-end of the siNA. In a non-limiting example, a double stranded siNA molecule of the invention in which each strand of the siNA is 21 nucleotides in length is featured wherein positions 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 and 21 of each strand are chemically modified (e.g., with compounds having any of Formulae I-VII, such as such as 2′-deoxy, 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy or 2′-O-methyl nucleotides). In another non-limiting example, a double stranded siNA molecule of the invention in which each strand of the siNA is 21 nucleotides in length is featured wherein positions 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 of each strand are chemically modified (e.g., with compounds having any of Formulae I-VII, such as such as 2′-deoxy, 2′-deoxy-2′-fluoro, 4′-thio, 2′-O-trifluoromethyl, 2′-O-ethyl-trifluoromethoxy, 2′-O-difluoromethoxy-ethoxy or 2′-O-methyl nucleotides). Such siNA molecules can further comprise terminal cap moieties and/or backbone modifications as described herein. 
     In one embodiment, the invention features a method for modulating the expression of a PTP-1B gene within a cell comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified or unmodified, wherein one of the siNA strands comprises a sequence complementary to RNA of the PTP-1B gene; and (b) introducing the siNA molecule into a cell under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B gene in the cell. 
     In one embodiment, the invention features a method for modulating the expression of a PTP-1B gene within a cell comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified or unmodified, wherein one of the siNA strands comprises a sequence complementary to RNA of the PTP-1B gene and wherein the sense strand sequence of the siNA comprises a sequence identical or substantially similar to the sequence of the target RNA; and (b) introducing the siNA molecule into a cell under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B gene in the cell. 
     In another embodiment, the invention features a method for modulating the expression of more than one PTP-1B gene within a cell comprising: (a) synthesizing siNA molecules of the invention, which can be chemically-modified or unmodified, wherein one of the siNA strands comprises a sequence complementary to RNA of the PTP-1B genes; and (b) introducing the siNA molecules into a cell under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B genes in the cell. 
     In another embodiment, the invention features a method for modulating the expression of two or more PTP-1B genes within a cell comprising: (a) synthesizing one or more siNA molecules of the invention, which can be chemically-modified or unmodified, wherein the siNA strands comprise sequences complementary to RNA of the PTP-1B genes and wherein the sense strand sequences of the siNAs comprise sequences identical or substantially similar to the sequences of the target RNAs; and (b) introducing the siNA molecules into a cell under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B genes in the cell. 
     In another embodiment, the invention features a method for modulating the expression of more than one PTP-1B gene within a cell comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified or unmodified, wherein one of the siNA strands comprises a sequence complementary to RNA of the PTP-1B gene and wherein the sense strand sequence of the siNA comprises a sequence identical or substantially similar to the sequences of the target RNAs; and (b) introducing the siNA molecule into a cell under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B genes in the cell. 
     In another embodiment, the invention features a method for modulating the expression of a PTP-1B gene within a cell comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified or unmodified, wherein one of the siNA strands comprises a sequence complementary to RNA of the PTP-1B gene, wherein the sense strand sequence of the siNA comprises a sequence identical or substantially similar to the sequences of the target RNA; and (b) introducing the siNA molecule into a cell under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B gene in the cell. 
     In one embodiment, siNA molecules of the invention are used as reagents in ex vivo applications. For example, siNA reagents are introduced into tissue or cells that are transplanted into a subject for therapeutic effect. The cells and/or tissue can be derived from an organism or subject that later receives the explant, or can be derived from another organism or subject prior to transplantation. The siNA molecules can be used to modulate the expression of one or more genes in the cells or tissue, such that the cells or tissue obtain a desired phenotype or are able to perform a function when transplanted in vivo. In one embodiment, certain target cells from a patient are extracted. These extracted cells are contacted with siNAs targeting a specific nucleotide sequence within the cells under conditions suitable for uptake of the siNAs by these cells (e.g. using delivery reagents such as cationic lipids, liposomes and the like or using techniques such as electroporation to facilitate the delivery of siNAs into cells). The cells are then reintroduced back into the same patient or other patients. 
     In one embodiment, the invention features a method of modulating the expression of a PTP-1B gene in a tissue explant comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified, wherein one of the siNA strands comprises a sequence complementary to RNA of the PTP-1B gene; and (b) introducing the siNA molecule into a cell of the tissue explant derived from a particular organism under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B gene in the tissue explant. In another embodiment, the method further comprises introducing the tissue explant back into the organism the tissue was derived from or into another organism under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B gene in that organism. 
     In one embodiment, the invention features a method of modulating the expression of a PTP-1B gene in a tissue explant comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified, wherein one of the siNA strands comprises a sequence complementary to RNA of the PTP-1B gene and wherein the sense strand sequence of the siNA comprises a sequence identical or substantially similar to the sequence of the target RNA; and (b) introducing the siNA molecule into a cell of the tissue explant derived from a particular organism under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B gene in the tissue explant. In another embodiment, the method further comprises introducing the tissue explant back into the organism the tissue was derived from or into another organism under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B gene in that organism. 
     In another embodiment, the invention features a method of modulating the expression of more than one PTP-1B gene in a tissue explant comprising: (a) synthesizing siNA molecules of the invention, which can be chemically-modified, wherein one of the siNA strands comprises a sequence complementary to RNA of the PTP-1B genes; and (b) introducing the siNA molecules into a cell of the tissue explant derived from a particular organism under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B genes in the tissue explant. In another embodiment, the method further comprises introducing the tissue explant back into the organism the tissue was derived from or into another organism under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B genes in that organism. 
     In one embodiment, the invention features a method of modulating the expression of a PTP-1B gene in a subject or organism comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified, wherein one of the siNA strands comprises a sequence complementary to RNA of the PTP-1B gene; and (b) introducing the siNA molecule into the subject or organism under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B gene in the subject or organism. The level of PTP-1B protein or RNA can be determined using various methods well-known in the art. 
     In another embodiment, the invention features a method of modulating the expression of more than one PTP-1 B gene in a subject or organism comprising: (a) synthesizing siNA molecules of the invention, which can be chemically-modified, wherein one of the siNA strands comprises a sequence complementary to RNA of the PTP-1B genes; and (b) introducing the siNA molecules into the subject or organism under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B genes in the subject or organism. The level of PTP-1B protein or RNA can be determined as is known in the art. 
     In one embodiment, the invention features a method for modulating the expression of a PTP-1B gene within a cell comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified, wherein the siNA comprises a single stranded sequence having complementarity to RNA of the PTP-1B gene; and (b) introducing the siNA molecule into a cell under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B gene in the cell. 
     In another embodiment, the invention features a method for modulating the expression of more than one PTP-1B gene within a cell comprising: (a) synthesizing siNA molecules of the invention, which can be chemically-modified, wherein the siNA comprises a single stranded sequence having complementarity to RNA of the PTP-1B gene; and (b) contacting the cell in vitro or in vivo with the siNA molecule under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B genes in the cell. 
     In one embodiment, the invention features a method of modulating the expression of a PTP-1B gene in a tissue explant (e.g., a skin, heart, liver, spleen, cornea, lung, stomach, kidney, vein, artery, hair, appendage, or limb transplant, or any other organ, tissue or cell as can be transplanted from one organism to another or back to the same organism from which the organ, tissue or cell is derived) comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified, wherein the siNA comprises a single stranded sequence having complementarity to RNA of the PTP-1B gene; and (b) contacting a cell of the tissue explant derived from a particular subject or organism with the siNA molecule under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B gene in the tissue explant. In another embodiment, the method further comprises introducing the tissue explant back into the subject or organism the tissue was derived from or into another subject or organism under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B gene in that subject or organism. 
     In another embodiment, the invention features a method of modulating the expression of more than one PTP-1B gene in a tissue explant (e.g., a skin, heart, liver, spleen, cornea, lung, stomach, kidney, vein, artery, hair, appendage, or limb transplant, or any other organ, tissue or cell as can be transplanted from one organism to another or back to the same organism from which the organ, tissue or cell is derived) comprising: (a) synthesizing siNA molecules of the invention, which can be chemically-modified, wherein the siNA comprises a single stranded sequence having complementarity to RNA of the PTP-1B gene; and (b) introducing the siNA molecules into a cell of the tissue explant derived from a particular subject or organism under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B genes in the tissue explant. In another embodiment, the method further comprises introducing the tissue explant back into the subject or organism the tissue was derived from or into another subject or organism under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B genes in that subject or organism. 
     In one embodiment, the invention features a method of modulating the expression of a PTP-1B gene in a subject or organism comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified, wherein the siNA comprises a single stranded sequence having complementarity to RNA of the PTP-1B gene; and (b) introducing the siNA molecule into the subject or organism under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B gene in the subject or organism. 
     In another embodiment, the invention features a method of modulating the expression of more than one PTP-1B gene in a subject or organism comprising: (a) synthesizing siNA molecules of the invention, which can be chemically-modified, wherein the siNA comprises a single stranded sequence having complementarity to RNA of the PTP-1B gene; and (b) introducing the siNA molecules into the subject or organism under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B genes in the subject or organism. 
     In one embodiment, the invention features a method of modulating the expression of a PTP-1B gene in a subject or organism comprising contacting the subject or organism with a siNA molecule of the invention under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B gene in the subject or organism. 
     In one embodiment, the invention features a method for treating or preventing diabetes (e.g., type 1 or type 2) in a subject or organism comprising contacting the subject or organism with a siNA molecule of the invention under conditions suitable to modulate the expression of the PTP-1B gene in the subject or organism whereby the treatment or prevention of diabetes can be achieved. In one embodiment, the invention features contacting the subject or organism with a siNA molecule of the invention via local administration to relevant tissues or cells, such as liver, intestine, or pancreas cells and tissues. In one embodiment, the invention features contacting the subject or organism with a siNA molecule of the invention via systemic administration (such as via intravenous or subcutaneous administration of siNA) to relevant tissues or cells, such as tissues or cells involved in the maintenance or development of diabetes. The siNA molecule of the invention can be formulated or conjugated as described herein or otherwise known in the art to target appropriate tissues or cells in the subject or organism. 
     In one embodiment, the invention features a method for treating or preventing obesity in a subject or organism comprising contacting the subject or organism with a siNA molecule of the invention under conditions suitable to modulate the expression of the PTP-1B gene in the subject or organism whereby the treatment or prevention of obesity can be achieved. In one embodiment, the invention features contacting the subject or organism with a siNA molecule of the invention via local administration to relevant tissues or cells, such as liver, intestine, or pancreas cells and tissues. In one embodiment, the invention features contacting the subject or organism with a siNA molecule of the invention via systemic administration (such as via intravenous or subcutaneous administration of siNA) to relevant tissues or cells, such as tissues or cells involved in the maintenance or development of obesity. The siNA molecule of the invention can be formulated or conjugated as described herein or otherwise known in the art to target appropriate tissues or cells in the subject or organism. 
     In one embodiment, the invention features a method for treating or preventing insulin resistance in a subject or organism comprising contacting the subject or organism with a siNA molecule of the invention under conditions suitable to modulate the expression of the PTP-1B gene in the subject or organism whereby the treatment or prevention of insulin resistance can be achieved. In one embodiment, the invention features contacting the subject or organism with a siNA molecule of the invention via local administration to relevant tissues or cells, such as liver, intestine, or pancreas cells and tissues. In one embodiment, the invention features contacting the subject or organism with a siNA molecule of the invention via systemic administration (such as via intravenous or subcutaneous administration of siNA) to relevant tissues or cells, such as tissues or cells involved in the maintenance or development of insulin resistance. The siNA molecule of the invention can be formulated or conjugated as described herein or otherwise known in the art to target appropriate tissues or cells in the subject or organism. 
     In any of the methods of treatment of the invention, the siNA can be administered to the subject as a course of treatment, for example administration at various time intervals, such as once per day over the course of treatment, once every two days over the course of treatment, once every three days over the course of treatment, once every four days over the course of treatment, once every five days over the course of treatment, once every six days over the course of treatment, once per week over the course of treatment, once every other week over the course of treatment, once per month over the course of treatment, etc. In one embodiment, the course of treatment is from about one to about 52 weeks or longer (e.g., indefinitely). In one embodiment, the course of treatment is from about one to about 48 months or longer (e.g., indefinitely). 
     In any of the methods of treatment of the invention, the siNA can be administered to the subject systemically as described herein or otherwise known in the art. Systemic administration can include, for example, intravenous, subcutaneous, intramuscular, catheterization, nasopharangeal, transdermal, or gastrointestinal administration as is generally known in the art. 
     In any of the methods of treatment of the invention, the siNA can be administered to the subject locally or to local tissues as described herein or otherwise known in the art. Local administration can include, for example, catheterization, implantation, direct injection, stenting, or portal vein administration to relevant tissues, or any other local administration technique, method or procedure, as is generally known in the art. 
     In another embodiment, the invention features a method of modulating the expression of more than one PTP-1B gene in a subject or organism comprising contacting the subject or organism with one or more siNA molecules of the invention under conditions suitable to modulate (e.g., inhibit) the expression of the PTP-1B genes in the subject or organism. 
     The siNA molecules of the invention can be designed to down regulate or inhibit target (e.g., PTP-1B) gene expression through RNAi targeting of a variety of nucleic acid molecules. In one embodiment, the siNA molecules of the invention are used to target various DNA corresponding to a target gene, for example via heterochromatic silencing. In one embodiment, the siNA molecules of the invention are used to target various RNAs corresponding to a target gene, for example via RNA target cleavage or translational inhibition. Non-limiting examples of such RNAs include messenger RNA (mRNA), non-coding RNA or regulatory elements, alternate RNA splice variants of target gene(s), post-transcriptionally modified RNA of target gene(s), pre-mRNA of target gene(s), and/or RNA templates. If alternate splicing produces a family of transcripts that are distinguished by usage of appropriate exons, the instant invention can be used to inhibit gene expression through the appropriate exons to specifically inhibit or to distinguish among the functions of gene family members. For example, a protein that contains an alternatively spliced transmembrane domain can be expressed in both membrane bound and secreted forms. Use of the invention to target the exon containing the transmembrane domain can be used to determine the functional consequences of pharmaceutical targeting of membrane bound as opposed to the secreted form of the protein. Non-limiting examples of applications of the invention relating to targeting these RNA molecules include therapeutic pharmaceutical applications, cosmetic applications, veterinary applications, pharmaceutical discovery applications, molecular diagnostic and gene function applications, and gene mapping, for example using single nucleotide polymorphism mapping with siNA molecules of the invention. Such applications can be implemented using known gene sequences or from partial sequences available from an expressed sequence tag (EST). 
     In another embodiment, the siNA molecules of the invention are used to target conserved sequences corresponding to a gene family or gene families such as PTP-1B family genes. As such, siNA molecules targeting multiple PTP-1B targets can provide increased therapeutic effect. In one embodiment, the invention features the targeting (cleavage or inhibition of expression or function) of more than one PTP-1B gene sequence using a single siNA molecule, by targeting the conserved sequences of the targeted PTP-1B gene. 
     In addition, siNA can be used to characterize pathways of gene function in a variety of applications. For example, the present invention can be used to inhibit the activity of target gene(s) in a pathway to determine the function of uncharacterized gene(s) in gene function analysis, mRNA function analysis, or translational analysis. The invention can be used to determine potential target gene pathways involved in various diseases and conditions toward pharmaceutical development. The invention can be used to understand pathways of gene expression involved in, for example, the progression and/or maintenance of diabetes (e.g., type I and/or type II), insulin resistance, obesity, and any other diseases, traits, and conditions associated with PTP-1B gene expression or activity in a subject or organism. 
     In one embodiment, siNA molecule(s) and/or methods of the invention are used to down regulate the expression of gene(s) that encode RNA referred to by Genbank Accession, for example, PTP-1B genes encoding RNA sequence(s) referred to herein by Genbank Accession number, for example, Genbank Accession Nos. shown in Table I. 
     In one embodiment, the invention features a method comprising: (a) generating a library of siNA constructs having a predetermined complexity; and (b) assaying the siNA constructs of (a) above, under conditions suitable to determine RNAi target sites within the target RNA sequence. In one embodiment, the siNA molecules of (a) have strands of a fixed length, for example, about 23 nucleotides in length. In another embodiment, the siNA molecules of (a) are of differing length, for example having strands of about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides in length. In one embodiment, the assay can comprise a reconstituted in vitro siNA assay as described herein. In another embodiment, the assay can comprise a cell culture system in which target RNA is expressed. In another embodiment, fragments of target RNA are analyzed for detectable levels of cleavage, for example by gel electrophoresis, northern blot analysis, or RNAse protection assays, to determine the most suitable target site(s) within the target RNA sequence. The target RNA sequence can be obtained as is known in the art, for example, by cloning and/or transcription for in vitro systems, and by cellular expression in in vivo systems. 
     In one embodiment, the invention features a method comprising: (a) generating a randomized library of siNA constructs having a predetermined complexity, such as of 4 N , where N represents the number of base paired nucleotides in each of the siNA construct strands (eg. for a siNA construct having 21 nucleotide sense and antisense strands with 19 base pairs, the complexity would be 4 19 ); and (b) assaying the siNA constructs of (a) above, under conditions suitable to determine RNAi target sites within the target PTP-1B RNA sequence. In another embodiment, the siNA molecules of (a) have strands of a fixed length, for example about 23 nucleotides in length. In yet another embodiment, the siNA molecules of (a) are of differing length, for example having strands of about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides in length. In one embodiment, the assay can comprise a reconstituted in vitro siNA assay as described in Example 6 herein. In another embodiment, the assay can comprise a cell culture system in which target RNA is expressed. In another embodiment, fragments of PTP-1B RNA are analyzed for detectable levels of cleavage, for example, by gel electrophoresis, northern blot analysis, or RNAse protection assays, to determine the most suitable target site(s) within the target PTP-1B RNA sequence. The target PTP-1B RNA sequence can be obtained as is known in the art, for example, by cloning and/or transcription for in vitro systems, and by cellular expression in in vivo systems. 
     In another embodiment, the invention features a method comprising: (a) analyzing the sequence of a RNA target encoded by a target gene; (b) synthesizing one or more sets of siNA molecules having sequence complementary to one or more regions of the RNA of (a); and (c) assaying the siNA molecules of (b) under conditions suitable to determine RNAi targets within the target RNA sequence. In one embodiment, the siNA molecules of (b) have strands of a fixed length, for example about 23 nucleotides in length. In another embodiment, the siNA molecules of (b) are of differing length, for example having strands of about 15 to about 30 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) nucleotides in length. In one embodiment, the assay can comprise a reconstituted in vitro siNA assay as described herein. In another embodiment, the assay can comprise a cell culture system in which target RNA is expressed. Fragments of target RNA are analyzed for detectable levels of cleavage, for example by gel electrophoresis, northern blot analysis, or RNAse protection assays, to determine the most suitable target site(s) within the target RNA sequence. The target RNA sequence can be obtained as is known in the art, for example, by cloning and/or transcription for in vitro systems, and by expression in in vivo systems. 
     By “target site” is meant a sequence within a target RNA that is “targeted” for cleavage mediated by a siNA construct which contains sequences within its antisense region that are complementary to the target sequence. 
     By “detectable level of cleavage” is meant cleavage of target RNA (and formation of cleaved product RNAs) to an extent sufficient to discern cleavage products above the background of RNAs produced by random degradation of the target RNA. Production of cleavage products from 1-5% of the target RNA is sufficient to detect above the background for most methods of detection. 
     In one embodiment, the invention features a composition comprising a siNA molecule of the invention, which can be chemically-modified, in a pharmaceutically acceptable carrier or diluent. In another embodiment, the invention features a pharmaceutical composition comprising siNA molecules of the invention, which can be chemically-modified, targeting one or more genes in a pharmaceutically acceptable carrier or diluent. In another embodiment, the invention features a method for diagnosing a disease, trait, or condition in a subject comprising administering to the subject a composition of the invention under conditions suitable for the diagnosis of the disease, trait, or condition in the subject. In another embodiment, the invention features a method for treating or preventing a disease, trait, or condition, such as diabetes (e.g., type I and/or type II), insulin resistance, or obesity in a subject, comprising administering to the subject a composition of the invention under conditions suitable for the treatment or prevention of the disease, trait, or condition in the subject, alone or in conjunction with one or more other therapeutic compounds. 
     In another embodiment, the invention features a method for validating a PTP-1B gene target, comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified, wherein one of the siNA strands includes a sequence complementary to RNA of a PTP-1B target gene; (b) introducing the siNA molecule into a cell, tissue, subject, or organism under conditions suitable for modulating expression of the PTP-1B target gene in the cell, tissue, subject, or organism; and (c) determining the function of the gene by assaying for any phenotypic change in the cell, tissue, subject, or organism. 
     In another embodiment, the invention features a method for validating a PTP-1B target comprising: (a) synthesizing a siNA molecule of the invention, which can be chemically-modified, wherein one of the siNA strands includes a sequence complementary to RNA of a PTP-1B target gene; (b) introducing the siNA molecule into a biological system under conditions suitable for modulating expression of the PTP-1B target gene in the biological system; and (c) determining the function of the gene by assaying for any phenotypic change in the biological system. 
     By “biological system” is meant, material, in a purified or unpurified form, from biological sources, including but not limited to human or animal, wherein the system comprises the components required for RNAi activity. The term “biological system” includes, for example, a cell, tissue, subject, or organism, or extract thereof. The term biological system also includes reconstituted RNAi systems that can be used in an in vitro setting. 
     By “phenotypic change” is meant any detectable change to a cell that occurs in response to contact or treatment with a nucleic acid molecule of the invention (e.g., siNA). Such detectable changes include, but are not limited to, changes in shape, size, proliferation, motility, protein expression or RNA expression or other physical or chemical changes as can be assayed by methods known in the art. The detectable change can also include expression of reporter genes/molecules such as Green Fluorescent Protein (GFP) or various tags that are used to identify an expressed protein or any other cellular component that can be assayed. 
     In one embodiment, the invention features a kit containing a siNA molecule of the invention, which can be chemically-modified, that can be used to modulate the expression of a PTP-1B target gene in a biological system, including, for example, in a cell, tissue, subject, or organism. In another embodiment, the invention features a kit containing more than one siNA molecule of the invention, which can be chemically-modified, that can be used to modulate the expression of more than one PTP-1B target gene in a biological system, including, for example, in a cell, tissue, subject, or organism. 
     In one embodiment, the invention features a cell containing one or more siNA molecules of the invention, which can be chemically-modified. In another embodiment, the cell containing a siNA molecule of the invention is a mammalian cell. In yet another embodiment, the cell containing a siNA molecule of the invention is a human cell. 
     In one embodiment, the synthesis of a siNA molecule of the invention, which can be chemically-modified, comprises: (a) synthesis of two complementary strands of the siNA molecule; (b) annealing the two complementary strands together under conditions suitable to obtain a double-stranded siNA molecule. In another embodiment, synthesis of the two complementary strands of the siNA molecule is by solid phase oligonucleotide synthesis. In yet another embodiment, synthesis of the two complementary strands of the siNA molecule is by solid phase tandem oligonucleotide synthesis. 
     In one embodiment, the invention features a method for synthesizing a siNA duplex molecule comprising: (a) synthesizing a first oligonucleotide sequence strand of the siNA molecule, wherein the first oligonucleotide sequence strand comprises a cleavable linker molecule that can be used as a scaffold for the synthesis of the second oligonucleotide sequence strand of the siNA; (b) synthesizing the second oligonucleotide sequence strand of siNA on the scaffold of the first oligonucleotide sequence strand, wherein the second oligonucleotide sequence strand further comprises a chemical moiety than can be used to purify the siNA duplex; (c) cleaving the linker molecule of (a) under conditions suitable for the two siNA oligonucleotide strands to hybridize and form a stable duplex; and (d) purifying the siNA duplex utilizing the chemical moiety of the second oligonucleotide sequence strand. In one embodiment, cleavage of the linker molecule in (c) above takes place during deprotection of the oligonucleotide, for example, under hydrolysis conditions using an alkylamine base such as methylamine. In one embodiment, the method of synthesis comprises solid phase synthesis on a solid support such as controlled pore glass (CPG) or polystyrene, wherein the first sequence of (a) is synthesized on a cleavable linker, such as a succinyl linker, using the solid support as a scaffold. The cleavable linker in (a) used as a scaffold for synthesizing the second strand can comprise similar reactivity as the solid support derivatized linker, such that cleavage of the solid support derivatized linker and the cleavable linker of (a) takes place concomitantly. In another embodiment, the chemical moiety of (b) that can be used to isolate the attached oligonucleotide sequence comprises a trityl group, for example a dimethoxytrityl group, which can be employed in a trityl-on synthesis strategy as described herein. In yet another embodiment, the chemical moiety, such as a dimethoxytrityl group, is removed during purification, for example, using acidic conditions. 
     In a further embodiment, the method for siNA synthesis is a solution phase synthesis or hybrid phase synthesis wherein both strands of the siNA duplex are synthesized in tandem using a cleavable linker attached to the first sequence which acts a scaffold for synthesis of the second sequence. Cleavage of the linker under conditions suitable for hybridization of the separate siNA sequence strands results in formation of the double-stranded siNA molecule. 
     In another embodiment, the invention features a method for synthesizing a siNA duplex molecule comprising: (a) synthesizing one oligonucleotide sequence strand of the siNA molecule, wherein the sequence comprises a cleavable linker molecule that can be used as a scaffold for the synthesis of another oligonucleotide sequence; (b) synthesizing a second oligonucleotide sequence having complementarity to the first sequence strand on the scaffold of (a), wherein the second sequence comprises the other strand of the double-stranded siNA molecule and wherein the second sequence further comprises a chemical moiety than can be used to isolate the attached oligonucleotide sequence; (c) purifying the product of (b) utilizing the chemical moiety of the second oligonucleotide sequence strand under conditions suitable for isolating the full-length sequence comprising both siNA oligonucleotide strands connected by the cleavable linker and under conditions suitable for the two siNA oligonucleotide strands to hybridize and form a stable duplex. In one embodiment, cleavage of the linker molecule in (c) above takes place during deprotection of the oligonucleotide, for example, under hydrolysis conditions. In another embodiment, cleavage of the linker molecule in (c) above takes place after deprotection of the oligonucleotide. In another embodiment, the method of synthesis comprises solid phase synthesis on a solid support such as controlled pore glass (CPG) or polystyrene, wherein the first sequence of (a) is synthesized on a cleavable linker, such as a succinyl linker, using the solid support as a scaffold. The cleavable linker in (a) used as a scaffold for synthesizing the second strand can comprise similar reactivity or differing reactivity as the solid support derivatized linker, such that cleavage of the solid support derivatized linker and the cleavable linker of (a) takes place either concomitantly or sequentially. In one embodiment, the chemical moiety of (b) that can be used to isolate the attached oligonucleotide sequence comprises a trityl group, for example a dimethoxytrityl group. 
     In another embodiment, the invention features a method for making a double-stranded siNA molecule in a single synthetic process comprising: (a) synthesizing an oligonucleotide having a first and a second sequence, wherein the first sequence is complementary to the second sequence, and the first oligonucleotide sequence is linked to the second sequence via a cleavable linker, and wherein a terminal 5′-protecting group, for example, a 5′-O-dimethoxytrityl group (5′-O-DMT) remains on the oligonucleotide having the second sequence; (b) deprotecting the oligonucleotide whereby the deprotection results in the cleavage of the linker joining the two oligonucleotide sequences; and (c) purifying the product of (b) under conditions suitable for isolating the double-stranded siNA molecule, for example using a trityl-on synthesis strategy as described herein. 
     In another embodiment, the method of synthesis of siNA molecules of the invention comprises the teachings of Scaringe et al., U.S. Pat. Nos. 5,889,136; 6,008,400; and 6,111,086, incorporated by reference herein in their entirety. 
     In one embodiment, the invention features siNA constructs that mediate RNAi against PTP-1B, wherein the siNA construct comprises one or more chemical modifications, for example, one or more chemical modifications having any of Formulae I-VII or any combination thereof that increases the nuclease resistance of the siNA construct. 
     In another embodiment, the invention features a method for generating siNA molecules with increased nuclease resistance comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having increased nuclease resistance. 
     In another embodiment, the invention features a method for generating siNA molecules with improved toxicologic profiles (e.g., having attenuated or no immunstimulatory properties) comprising (a) introducing nucleotides having any of Formula I-VII (e.g., siNA motifs referred to in Table IV) or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved toxicologic profiles. 
     In another embodiment, the invention features a method for generating siNA formulations with improved toxicologic profiles (e.g., having attenuated or no immunstimulatory properties) comprising (a) generating a siNA formulation comprising a siNA molecule of the invention and a delivery vehicle or delivery particle as described herein or as otherwise known in the art, and (b) assaying the siNA formulation of step (a) under conditions suitable for isolating siNA formulations having improved toxicologic profiles. 
     In another embodiment, the invention features a method for generating siNA molecules that do not stimulate an interferon response (e.g., no interferon response or attenuated interferon response) in a cell, subject, or organism, comprising (a) introducing nucleotides having any of Formula I-VII (e.g., siNA motifs referred to in Table IV) or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules that do not stimulate an interferon response. 
     In another embodiment, the invention features a method for generating siNA formulations that do not stimulate an interferon response (e.g., no interferon response or attenuated interferon response) in a cell, subject, or organism, comprising (a) generating a siNA formulation comprising a siNA molecule of the invention and a delivery vehicle or delivery particle as described herein or as otherwise known in the art, and (b) assaying the siNA formulation of step (a) under conditions suitable for isolating siNA formulations that do not stimulate an interferon response. 
     By “improved toxicologic profile”, is meant that the chemically modified or formulated siNA construct exhibits decreased toxicity in a cell, subject, or organism compared to an unmodified or unformulated siNA, or siNA molecule having fewer modifications or modifications that are less effective in imparting improved toxicology. In a non-limiting example, siNA molecules and formulations with improved toxicologic profiles are associated with a decreased or attenuated immunostimulatory response in a cell, subject, or organism compared to an unmodified or unformulated siNA, or siNA molecule having fewer modifications or modifications that are less effective in imparting improved toxicology. In one embodiment, a siNA molecule or formulation with an improved toxicological profile comprises no ribonucleotides. In one embodiment, a siNA molecule or formulation with an improved toxicological profile comprises less than 5 ribonucleotides (e.g., 1, 2, 3, or 4 ribonucleotides). In one embodiment, a siNA molecule or formulation with an improved toxicological profile comprises Stab 7, Stab 8, Stab 11, Stab 12, Stab 13, Stab 16, Stab 17, Stab 18, Stab 19, Stab 20, Stab 23, Stab 24, Stab 25, Stab 26, Stab 27, Stab 28, Stab 29, Stab 30, Stab 31, Stab 32, Stab 33, Stab 34 or any combination thereof (see Table IV). Herein, numeric Stab chemistries include both 2′-fluoro and 2′-OCF3 versions of the chemistries shown in Table IV. For example, “Stab 7/8” refers to both Stab 7/8 and Stab 7F/8F etc. In one embodiment, a siNA molecule or formulation with an improved toxicological profile comprises a siNA molecule of the invention and a formulation as described in United States Patent Application Publication No. 20030077829, incorporated by reference herein in its entirety including the drawings. In one embodiment, the level of immunostimulatory response associated with a given siNA molecule can be measured as is known in the art, for example by determining the level of PKR/interferon response, proliferation, B-cell activation, and/or cytokine production in assays to quantitate the immunostimulatory response of particular siNA molecules (see, for example, Leifer et al., 2003,  J Immunother.  26, 313-9; and U.S. Pat. No. 5,968,909, incorporated in its entirety by reference). 
     In one embodiment, the invention features siNA constructs that mediate RNAi against PTP-IB, wherein the siNA construct comprises one or more chemical modifications described herein that modulates the binding affinity between the sense and antisense strands of the siNA construct. 
     In another embodiment, the invention features a method for generating siNA molecules with increased binding affinity between the sense and antisense strands of the siNA molecule comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having increased binding affinity between the sense and antisense strands of the siNA molecule. 
     In one embodiment, the invention features siNA constructs that mediate RNAi against PTP-1B, wherein the siNA construct comprises one or more chemical modifications described herein that modulates the binding affinity between the antisense strand of the siNA construct and a complementary target RNA sequence within a cell. 
     In one embodiment, the invention features siNA constructs that mediate RNAi against PTP-1B, wherein the siNA construct comprises one or more chemical modifications described herein that modulates the binding affinity between the antisense strand of the siNA construct and a complementary target DNA sequence within a cell. 
     In another embodiment, the invention features a method for generating siNA molecules with increased binding affinity between the antisense strand of the siNA molecule and a complementary target RNA sequence comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having increased binding affinity between the antisense strand of the siNA molecule and a complementary target RNA sequence. 
     In another embodiment, the invention features a method for generating siNA molecules with increased binding affinity between the antisense strand of the siNA molecule and a complementary target DNA sequence comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having increased binding affinity between the antisense strand of the siNA molecule and a complementary target DNA sequence. 
     In one embodiment, the invention features siNA constructs that mediate RNAi against PTP-1B, wherein the siNA construct comprises one or more chemical modifications described herein that modulate the polymerase activity of a cellular polymerase capable of generating additional endogenous siNA molecules having sequence homology to the chemically-modified siNA construct. 
     In another embodiment, the invention features a method for generating siNA molecules capable of mediating increased polymerase activity of a cellular polymerase capable of generating additional endogenous siNA molecules having sequence homology to a chemically-modified siNA molecule comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules capable of mediating increased polymerase activity of a cellular polymerase capable of generating additional endogenous siNA molecules having sequence homology to the chemically-modified siNA molecule. 
     In one embodiment, the invention features chemically-modified siNA constructs that mediate RNAi against PTP-1B in a cell, wherein the chemical modifications do not significantly effect the interaction of siNA with a target RNA molecule, DNA molecule and/or proteins or other factors that are essential for RNAi in a manner that would decrease the efficacy of RNAi mediated by such siNA constructs. 
     In another embodiment, the invention features a method for generating siNA molecules with improved RNAi specificity against PTP-1B targets comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved RNAi specificity. In one embodiment, improved specificity comprises having reduced off target effects compared to an unmodified siNA molecule. For example, introduction of terminal cap moieties at the 3′-end, 5′-end, or both 3′ and 5′-ends of the sense strand or region of a siNA molecule of the invention can direct the siNA to have improved specificity by preventing the sense strand or sense region from acting as a template for RNAi activity against a corresponding target having complementarity to the sense strand or sense region. 
     In another embodiment, the invention features a method for generating siNA molecules with improved RNAi activity against PTP-1B comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved RNAi activity. 
     In yet another embodiment, the invention features a method for generating siNA molecules with improved RNAi activity against PTP-1B target RNA comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved RNAi activity against the target RNA. 
     In yet another embodiment, the invention features a method for generating siNA molecules with improved RNAi activity against PTP-1B target DNA comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved RNAi activity against the target DNA. 
     In one embodiment, the invention features siNA constructs that mediate RNAi against PTP-1B, wherein the siNA construct comprises one or more chemical modifications described herein that modulates the cellular uptake of the siNA construct, such as cholesterol conjugation of the siNA. 
     In another embodiment, the invention features a method for generating siNA molecules against PTP-1B with improved cellular uptake comprising (a) introducing nucleotides having any of Formula I-VII or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved cellular uptake. 
     In one embodiment, the invention features siNA constructs that mediate RNAi against PTP-1B, wherein the siNA construct comprises one or more chemical modifications described herein that increases the bioavailability of the siNA construct, for example, by attaching polymeric conjugates such as polyethyleneglycol or equivalent conjugates that improve the pharmacokinetics of the siNA construct, or by attaching conjugates that target specific tissue types or cell types in vivo. Non-limiting examples of such conjugates are described in Vargeese et al., U.S. Ser. No. 10/201,394 incorporated by reference herein. 
     In one embodiment, the invention features a method for generating siNA molecules of the invention with improved bioavailability comprising (a) introducing a conjugate into the structure of a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved bioavailability. Such conjugates can include ligands for cellular receptors, such as peptides derived from naturally occurring protein ligands; protein localization sequences, including cellular ZIP code sequences; antibodies; nucleic acid aptamers; vitamins and other co-factors, such as folate and N-acetylgalactosamine; polymers, such as polyethyleneglycol (PEG); phospholipids; cholesterol; cholesterol derivatives, polyamines, such as spermine or spermidine; and others. 
     In one embodiment, the invention features a double stranded short interfering nucleic acid (siNA) molecule that comprises a first nucleotide sequence complementary to a target RNA sequence or a portion thereof, and a second sequence having complementarity to said first sequence, wherein said second sequence is chemically modified in a manner that it can no longer act as a guide sequence for efficiently mediating RNA interference and/or be recognized by cellular proteins that facilitate RNAi. In one embodiment, the first nucleotide sequence of the siNA is chemically modified as described herein. In one embodiment, the first nucleotide sequence of the siNA is not modified (e.g., is all RNA). 
     In one embodiment, the invention features a double stranded short interfering nucleic acid (siNA) molecule that comprises a first nucleotide sequence complementary to a target RNA sequence or a portion thereof, and a second sequence having complementarity to said first sequence, wherein the second sequence is designed or modified in a manner that prevents its entry into the RNAi pathway as a guide sequence or as a sequence that is complementary to a target nucleic acid (e.g., RNA) sequence. In one embodiment, the first nucleotide sequence of the siNA is chemically modified as described herein. In one embodiment, the first nucleotide sequence of the siNA is not modified (e.g., is all RNA). Such design or modifications are expected to enhance the activity of siNA and/or improve the specificity of siNA molecules of the invention. These modifications are also expected to minimize any off-target effects and/or associated toxicity. 
     In one embodiment, the invention features a double stranded short interfering nucleic acid (siNA) molecule that comprises a first nucleotide sequence complementary to a target RNA sequence or a portion thereof, and a second sequence having complementarity to said first sequence, wherein said second sequence is incapable of acting as a guide sequence for mediating RNA interference. In one embodiment, the first nucleotide sequence of the siNA is chemically modified as described herein. In one embodiment, the first nucleotide sequence of the siNA is not modified (e.g., is all RNA). 
     In one embodiment, the invention features a double stranded short interfering nucleic acid (siNA) molecule that comprises a first nucleotide sequence complementary to a target RNA sequence or a portion thereof, and a second sequence having complementarity to said first sequence, wherein said second sequence does not have a terminal 5′-hydroxyl (5′-OH) or 5′-phosphate group. 
     In one embodiment, the invention features a double stranded short interfering nucleic acid (siNA) molecule that comprises a first nucleotide sequence complementary to a target RNA sequence or a portion thereof, and a second sequence having complementarity to said first sequence, wherein said second sequence comprises a terminal cap moiety at the 5′-end of said second sequence. In one embodiment, the terminal cap moiety comprises an inverted abasic, inverted deoxy abasic, inverted nucleotide moiety, a group shown in  FIG. 10 , an alkyl or cycloalkyl group, a heterocycle, or any other group that prevents RNAi activity in which the second sequence serves as a guide sequence or template for RNAi. 
     In one embodiment, the invention features a double stranded short interfering nucleic acid (siNA) molecule that comprises a first nucleotide sequence complementary to a target RNA sequence or a portion thereof, and a second sequence having complementarity to said first sequence, wherein said second sequence comprises a terminal cap moiety at the 5′-end and 3′-end of said second sequence. In one embodiment, each terminal cap moiety individually comprises an inverted abasic, inverted deoxy abasic, inverted nucleotide moiety, a group shown in  FIG. 10 , an alkyl or cycloalkyl group, a heterocycle, or any other group that prevents RNAi activity in which the second sequence serves as a guide sequence or template for RNAi. 
     In one embodiment, the invention features a method for generating siNA molecules of the invention with improved specificity for down regulating or inhibiting the expression of a target nucleic acid (e.g., a DNA or RNA such as a gene or its corresponding RNA), comprising (a) introducing one or more chemical modifications into the structure of a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved specificity. In another embodiment, the chemical modification used to improve specificity comprises terminal cap modifications at the 5′-end, 3′-end, or both 5′ and 3′-ends of the siNA molecule. The terminal cap modifications can comprise, for example, structures shown in  FIG. 10  (e.g. inverted deoxyabasic moieties) or any other chemical modification that renders a portion of the siNA molecule (e.g. the sense strand) incapable of mediating RNA interference against an off target nucleic acid sequence. In a non-limiting example, a siNA molecule is designed such that only the antisense sequence of the siNA molecule can serve as a guide sequence for RISC mediated degradation of a corresponding target RNA sequence. This can be accomplished by rendering the sense sequence of the siNA inactive by introducing chemical modifications to the sense strand that preclude recognition of the sense strand as a guide sequence by RNAi machinery. In one embodiment, such chemical modifications comprise any chemical group at the 5′-end of the sense strand of the siNA, or any other group that serves to render the sense strand inactive as a guide sequence for mediating RNA interference. These modifications, for example, can result in a molecule where the 5′-end of the sense strand no longer has a free 5′-hydroxyl (5′-OH) or a free 5′-phosphate group (e.g., phosphate, diphosphate, triphosphate, cyclic phosphate etc.). Non-limiting examples of such siNA constructs are described herein, such as “Stab 9/10”, “Stab 7/8”, “Stab 7/19”, “Stab 17/22”, “Stab 23/24”, “Stab 24/25”, and “Stab 24/26” (e.g., any siNA having Stab 7, 9, 17, 23, or 24 sense strands) chemistries and variants thereof (see Table IV) wherein the 5′-end and 3′-end of the sense strand of the siNA do not comprise a hydroxyl group or phosphate group. Herein, numeric Stab chemistries include both 2′-fluoro and 2′-OCF3 versions of the chemistries shown in Table IV. For example, “Stab 7/8” refers to both Stab 7/8 and Stab 7F/8F etc. 
     In one embodiment, the invention features a method for generating siNA molecules of the invention with improved specificity for down regulating or inhibiting the expression of a target nucleic acid (e.g., a DNA or RNA such as a gene or its corresponding RNA), comprising introducing one or more chemical modifications into the structure of a siNA molecule that prevent a strand or portion of the siNA molecule from acting as a template or guide sequence for RNAi activity. In one embodiment, the inactive strand or sense region of the siNA molecule is the sense strand or sense region of the siNA molecule, i.e. the strand or region of the siNA that does not have complementarity to the target nucleic acid sequence. In one embodiment, such chemical modifications comprise any chemical group at the 5′-end of the sense strand or region of the siNA that does not comprise a 5′-hydroxyl (5′-OH) or 5′-phosphate group, or any other group that serves to render the sense strand or sense region inactive as a guide sequence for mediating RNA interference. Non-limiting examples of such siNA constructs are described herein, such as “Stab 9/10”, “Stab 7/8”, “Stab 7/19”, “Stab 17/22”, “Stab 23/24”, “Stab 24/25”, and “Stab 24/26” (e.g., any siNA having Stab 7, 9, 17, 23, or 24 sense strands) chemistries and variants thereof (see Table IV) wherein the 5′-end and 3′-end of the sense strand of the siNA do not comprise a hydroxyl group or phosphate group. Herein, numeric Stab chemistries include both 2′-fluoro and 2′-OCF3 versions of the chemistries shown in Table IV. For example, “Stab 7/8” refers to both Stab 7/8 and Stab 7F/8F etc. 
     In one embodiment, the invention features a method for screening siNA molecules that are active in mediating RNA interference against a target nucleic acid sequence comprising (a) generating a plurality of unmodified siNA molecules, (b) screening the siNA molecules of step (a) under conditions suitable for isolating siNA molecules that are active in mediating RNA interference against the target nucleic acid sequence, and (c) introducing chemical modifications (e.g. chemical modifications as described herein or as otherwise known in the art) into the active siNA molecules of (b). In one embodiment, the method further comprises re-screening the chemically modified siNA molecules of step (c) under conditions suitable for isolating chemically modified siNA molecules that are active in mediating RNA interference against the target nucleic acid sequence. 
     In one embodiment, the invention features a method for screening chemically modified siNA molecules that are active in mediating RNA interference against a target nucleic acid sequence comprising (a) generating a plurality of chemically modified siNA molecules (e.g. siNA molecules as described herein or as otherwise known in the art), and (b) screening the siNA molecules of step (a) under conditions suitable for isolating chemically modified siNA molecules that are active in mediating RNA interference against the target nucleic acid sequence. 
     The term “ligand” refers to any compound or molecule, such as a drug, peptide, hormone, or neurotransmitter, that is capable of interacting with another compound, such as a receptor, either directly or indirectly. The receptor that interacts with a ligand can be present on the surface of a cell or can alternately be an intercellular receptor. Interaction of the ligand with the receptor can result in a biochemical reaction, or can simply be a physical interaction or association. 
     In another embodiment, the invention features a method for generating siNA molecules of the invention with improved bioavailability comprising (a) introducing an excipient formulation to a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved bioavailability. Such excipients include polymers such as cyclodextrins, lipids, cationic lipids, polyamines, phospholipids, nanoparticles, receptors, ligands, and others. 
     In another embodiment, the invention features a method for generating siNA molecules of the invention with improved bioavailability comprising (a) introducing nucleotides having any of Formulae or any combination thereof into a siNA molecule, and (b) assaying the siNA molecule of step (a) under conditions suitable for isolating siNA molecules having improved bioavailability. 
     In another embodiment, polyethylene glycol (PEG) can be covalently attached to siNA compounds of the present invention. The attached PEG can be any molecular weight, preferably from about 100 to about 50,000 daltons (Da). 
     The present invention can be used alone or as a component of a kit having at least one of the reagents necessary to carry out the in vitro or in vivo introduction of RNA to test samples and/or subjects. For example, preferred components of the kit include a siNA molecule of the invention and a vehicle that promotes introduction of the siNA into cells of interest as described herein (e.g., using lipids and other methods of transfection known in the art, see for example Beigelman et al, U.S. Pat. No. 6,395,713). The kit can be used for target validation, such as in determining gene function and/or activity, or in drug optimization, and in drug discovery (see for example Usman et al., U.S. Ser. No. 60/402,996). Such a kit can also include instructions to allow a user of the kit to practice the invention. 
     The term “short interfering nucleic acid”, “siNA”, “short interfering RNA”, “siRNA”, “short interfering nucleic acid molecule”, “short interfering oligonucleotide molecule”, or “chemically-modified short interfering nucleic acid molecule” as used herein refers to any nucleic acid molecule capable of inhibiting or down regulating gene expression or viral replication, for example by mediating RNA interference “RNAi” or gene silencing in a sequence-specific manner; see for example Zamorc et al., 2000, Cell, 101, 25-33; Bass, 2001,  Nature,  411, 428-429; Elbashir et al., 2001,  Nature,  411, 494-498; and Kreutzer et al., International PCT Publication No. WO 00/44895; Zernicka-Goetz et al., International PCT Publication No. WO 01/36646; Fire, International PCT Publication No. WO 99/32619; Plaetinck et al., International PCT Publication No. WO 00/01846; Mello and Fire, International PCT Publication No. WO 01/29058; Deschamps-Depaillette, International PCT Publication No. WO 99/07409; and Li et al., International PCT Publication No. WO 00/44914; Allshire, 2002,  Science,  297, 1818-1819; Volpe et al., 2002,  Science,  297, 1833-1837; Jenuwein, 2002,  Science,  297, 2215-2218; and Hall et al., 2002,  Science,  297, 2232-2237; Hutvagner and Zamore, 2002,  Science,  297, 2056-60; McManus et al., 2002,  RNA,  8, 842-850; Reinhart et al., 2002,  Gene  &amp;  Dev.,  16, 1616-1626; and Reinhart &amp; Bartel, 2002,  Science,  297, 1831). Non limiting examples of siNA molecules of the invention are shown in  FIGS. 4-6 , and Tables II and III herein. For example the siNA can be a double-stranded polynucleotide molecule comprising self-complementary sense and antisense regions, wherein the antisense region comprises nucleotide sequence that is complementary to nucleotide sequence in a target nucleic acid molecule or a portion thereof and the sense region having nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof. The siNA can be assembled from two separate oligonucleotides, where one strand is the sense strand and the other is the antisense strand, wherein the antisense and sense strands are self-complementary (i.e., each strand comprises nucleotide sequence that is complementary to nucleotide sequence in the other strand; such as where the antisense strand and sense strand form a duplex or double stranded structure, for example wherein the double stranded region is about 15 to about 30, e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 base pairs; the antisense strand comprises nucleotide sequence that is complementary to nucleotide sequence in a target nucleic acid molecule or a portion thereof and the sense strand comprises nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof (e.g., about 15 to about 25 or more nucleotides of the siNA molecule are complementary to the target nucleic acid or a portion thereof). Alternatively, the siNA is assembled from a single oligonucleotide, where the self-complementary sense and antisense regions of the siNA are linked by means of a nucleic acid based or non-nucleic acid-based linker(s). The siNA can be a polynucleotide with a duplex, asymmetric duplex, hairpin or asymmetric hairpin secondary structure, having self-complementary sense and antisense regions, wherein the antisense region comprises nucleotide sequence that is complementary to nucleotide sequence in a separate target nucleic acid molecule or a portion thereof and the sense region having nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof. The siNA can be a circular single-stranded polynucleotide having two or more loop structures and a stem comprising self-complementary sense and antisense regions, wherein the antisense region comprises nucleotide sequence that is complementary to nucleotide sequence in a target nucleic acid molecule or a portion thereof and the sense region having nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof, and wherein the circular polynucleotide can be processed either in vivo or in vitro to generate an active siNA molecule capable of mediating RNAi. The siNA can also comprise a single stranded polynucleotide having nucleotide sequence complementary to nucleotide sequence in a target nucleic acid molecule or a portion thereof (for example, where such siNA molecule does not require the presence within the siNA molecule of nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof), wherein the single stranded polynucleotide can further comprise a terminal phosphate group, such as a 5′-phosphate (see for example Martinez et al., 2002,  Cell.,  110, 563-574 and Schwarz et al., 2002,  Molecular Cell,  10, 537-568), or 5′,3′-diphosphate. In certain embodiments, the siNA molecule of the invention comprises separate sense and antisense sequences or regions, wherein the sense and antisense regions are covalently linked by nucleotide or non-nucleotide linkers molecules as is known in the art, or are alternately non-covalently linked by ionic interactions, hydrogen bonding, van der waals interactions, hydrophobic interactions, and/or stacking interactions. In certain embodiments, the siNA molecules of the invention comprise nucleotide sequence that is complementary to nucleotide sequence of a target gene. In another embodiment, the siNA molecule of the invention interacts with nucleotide sequence of a target gene in a manner that causes inhibition of expression of the target gene. As used herein, siNA molecules need not be limited to those molecules containing only RNA, but further encompasses chemically-modified nucleotides and non-nucleotides. In certain embodiments, the short interfering nucleic acid molecules of the invention lack 2′-hydroxy (2′-OH) containing nucleotides. Applicant describes in certain embodiments short interfering nucleic acids that do not require the presence of nucleotides having a 2′-hydroxy group for mediating RNAi and as such, short interfering nucleic acid molecules of the invention optionally do not include any ribonucleotides (e.g., nucleotides having a 2′-OH group). Such siNA molecules that do not require the presence of ribonucleotides within the siNA molecule to support RNAi can however have an attached linker or linkers or other attached or associated groups, moieties, or chains containing one or more nucleotides with 2′-OH groups. Optionally, siNA molecules can comprise ribonucleotides at about 5, 10, 20, 30, 40, or 50% of the nucleotide positions. The modified short interfering nucleic acid molecules of the invention can also be referred to as short interfering modified oligonucleotides “siMON.” As used herein, the term siNA is meant to be equivalent to other terms used to describe nucleic acid molecules that are capable of mediating sequence specific RNAi, for example short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), short hairpin RNA (shRNA), short interfering oligonucleotide, short interfering nucleic acid, short interfering modified oligonucleotide, chemically-modified siRNA, post-transcriptional gene silencing RNA (ptgsRNA), and others. In addition, as used herein, the term RNAi is meant to be equivalent to other terms used to describe sequence specific RNA interference, such as post transcriptional gene silencing, translational inhibition, or epigenetics. For example, siNA molecules of the invention can be used to epigenetically silence genes at both the post-transcriptional level or the pre-transcriptional level. In a non-limiting example, epigenetic modulation of gene expression by siNA molecules of the invention can result from siNA mediated modification of chromatin structure or methylation pattern to alter gene expression (see, for example, Verdel et al., 2004,  Science,  303, 672-676; Pal-Bhadra et al., 2004,  Science,  303, 669-672; Allshire, 2002,  Science,  297, 1818-1819; Volpe et al., 2002,  Science,  297, 1833-1837; Jenuwein, 2002,  Science,  297, 2215-2218; and Hall et al., 2002,  Science,  297, 2232-2237). In another non-limiting example, modulation of gene expression by siNA molecules of the invention can result from siNA mediated cleavage of RNA (either coding or non-coding RNA) via RISC, or alternately, translational inhibition as is known in the art. 
     In one embodiment, a siNA molecule of the invention is a duplex forming oligonucleotide “DFO”, (see for example  FIGS. 14-15  and Vaish et al., U.S. Ser. No. 10/727,780 filed Dec. 3, 2003 and International PCT Application No. US04/16390, filed May 24, 2004). 
     In one embodiment, a siNA molecule of the invention is a multifunctional siNA, (see for example  FIGS. 16-21  and Jadhav et al., U.S. Ser. No. 60/543,480 filed Feb. 10, 2004 and International PCT Application No. US04/16390, filed May 24, 2004). In one embodiment, the multifunctional siNA of the invention can comprise sequence targeting, for example, two or more regions of PTP-1B RNA (see for example target sequences in Tables II and III). 
     By “asymmetric hairpin” as used herein is meant a linear siNA molecule comprising an antisense region, a loop portion that can comprise nucleotides or non-nucleotides, and a sense region that comprises fewer nucleotides than the antisense region to the extent that the sense region has enough complementary nucleotides to base pair with the antisense region and form a duplex with loop. For example, an asymmetric hairpin siNA molecule of the invention can comprise an antisense region having length sufficient to mediate RNAi in a cell or in vitro system (e.g. about 15 to about 30, or about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides) and a loop region comprising about 4 to about 12 (e.g., about 4, 5, 6, 7, 8, 9, 10, 11, or 12) nucleotides, and a sense region having about 3 to about 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) nucleotides that are complementary to the antisense region. The asymmetric hairpin siNA molecule can also comprise a 5′-terminal phosphate group that can be chemically modified. The loop portion of the asymmetric hairpin siNA molecule can comprise nucleotides, non-nucleotides, linker molecules, or conjugate molecules as described herein. 
     By “asymmetric duplex” as used herein is meant a siNA molecule having two separate strands comprising a sense region and an antisense region, wherein the sense region comprises fewer nucleotides than the antisense region to the extent that the sense region has enough complementary nucleotides to base pair with the antisense region and form a duplex. For example, an asymmetric duplex siNA molecule of the invention can comprise an antisense region having length sufficient to mediate RNAi in a cell or in vitro system (e.g., about 15 to about 30, or about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides) and a sense region having about 3 to about 25 (e.g., about 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) nucleotides that are complementary to the antisense region. 
     By “modulate” is meant that the expression of the gene, or level of a RNA molecule or equivalent RNA molecules encoding one or more proteins or protein subunits, or activity of one or more proteins or protein subunits is up regulated or down regulated, such that expression, level, or activity is greater than or less than that observed in the absence of the modulator. For example, the term “modulate” can mean “inhibit,” but the use of the word “modulate” is not limited to this definition. 
     By “inhibit”, “down-regulate”, or “reduce”, it is meant that the expression of the gene, or level of RNA molecules or equivalent RNA molecules encoding one or more proteins or protein subunits, or activity of one or more proteins or protein subunits, is reduced below that observed in the absence of the nucleic acid molecules (e.g., siNA) of the invention. In one embodiment, inhibition, down-regulation or reduction with an siNA molecule is below that level observed in the presence of an inactive or attenuated molecule. In another embodiment, inhibition, down-regulation, or reduction with siNA molecules is below that level observed in the presence of, for example, an siNA molecule with scrambled sequence or with mismatches. In another embodiment, inhibition, down-regulation, or reduction of gene expression with a nucleic acid molecule of the instant invention is greater in the presence of the nucleic acid molecule than in its absence. In one embodiment, inhibition, down regulation, or reduction of gene expression is associated with post transcriptional silencing, such as RNAi mediated cleavage of a target nucleic acid molecule (e.g. RNA) or inhibition of translation. In one embodiment, inhibition, down regulation, or reduction of gene expression is associated with pretranscriptional silencing, such as by alterations in DNA methylation patterns and DNA chromatin structure. 
     By “gene”, or “target gene”, is meant a nucleic acid that encodes an RNA, for example, nucleic acid sequences including, but not limited to, structural genes encoding a polypeptide. A gene or target gene can also encode a functional RNA (fRNA) or non-coding RNA (ncRNA), such as small temporal RNA (stRNA), micro RNA (miRNA), small nuclear RNA (snRNA), short interfering RNA (siRNA), small nucleolar RNA (snRNA), ribosomal RNA (rRNA), transfer RNA (tRNA) and precursor RNAs thereof. Such non-coding RNAs can serve as target nucleic acid molecules for siNA mediated RNA interference in modulating the activity of fRNA or ncRNA involved in functional or regulatory cellular processes. Abberant fRNA or ncRNA activity leading to disease can therefore be modulated by siNA molecules of the invention. siNA molecules targeting fRNA and ncRNA can also be used to manipulate or alter the genotype or phenotype of a subject, organism or cell, by intervening in cellular processes such as genetic imprinting, transcription, translation, or nucleic acid processing (e.g., transamination, methylation etc.). The target gene can be a gene derived from a cell, an endogenous gene, a transgene, or exogenous genes such as genes of a pathogen, for example a virus, which is present in the cell after infection thereof. The cell containing the target gene can be derived from or contained in any organism, for example a plant, animal, protozoan, virus, bacterium, or fungus. Non-limiting examples of plants include monocots, dicots, or gymnosperms. Non-limiting examples of animals include vertebrates or invertebrates. Non-limiting examples of fungi include molds or yeasts. For a review, see for example Snyder and Gerstein, 2003,  Science,  300, 258-260. 
     By “non-canonical base pair” is meant any non-Watson Crick base pair, such as mismatches and/or wobble base pairs, including flipped mismatches, single hydrogen bond mismatches, trans-type mismatches, triple base interactions, and quadruple base interactions. Non-limiting examples of such non-canonical base pairs include, but are not limited to, AC reverse Hoogsteen, AC wobble, AU reverse Hoogsteen, GU wobble, AA N7 amino, CC 2-carbonyl-amino(H1)-N3-amino(H2), GA sheared, UC 4-carbonyl-amino, UU imino-carbonyl, AC reverse wobble, AU Hoogsteen, AU reverse Watson Crick, CG reverse Watson Crick, GC N3-amino-amino N3, AA N1-amino symmetric, AA N7-amino symmetric, GA N7-N1 amino-carbonyl, GA+ carbonyl-amino N7-N1, GG N1-carbonyl symmetric, GG N3-amino symmetric, CC carbonyl-amino symmetric, CC N3-amino symmetric, UU 2-carbonyl-imino symmetric, UU 4-carbonyl-imino symmetric, AA amino-N3, AA N1-amino, AC amino 2-carbonyl, AC N3-amino, AC N7-amino, AU amino-4-carbonyl, AU N1-imino, AU N3-imino, AU N7-imino, CC carbonyl-amino, GA amino-N1, GA amino-N7, GA carbonyl-amino, GA N3-amino, GC amino-N3, GC carbonyl-amino, GC N3-amino, GC N7-amino, GG amino-N7, GG carbonyl-imino, GG N7-amino, GU amino-2-carbonyl, GU carbonyl-imino, GU imino-2-carbonyl, GU N7-imino, psiU imino-2-carbonyl, UC 4-carbonyl-amino, UC imino-carbonyl, UU imino-4-carbonyl, AC C2-H-N3, GA carbonyl-C2-H, UU imino-4-carbonyl 2 carbonyl-C5-H, AC amino(A) N3(C)-carbonyl, GC imino amino-carbonyl, Gpsi imino-2-carbonyl amino-2-carbonyl, and GU imino amino-2-carbonyl base pairs. 
     By “protein tyrosine phosphatase-1B”, “PTPN1” or “PTP-1B” as used herein is meant, PTP-1B protein, peptide, or polypeptide having PTP-1B activity, such as encoded by PTP-1B Genbank Accession Nos. shown in Table I. The term PTP-1B also refers to nucleic acid sequences encoding any PTP-1B protein, peptide, or polypeptide having PTP-1B activity. The term “PTP-1B” is also meant to include other PTP-1B encoding sequence, such as other protein tyrosine phosphatase isoforms (e.g., protein tyrosine phosphatase alpha, etc.), mutant PTP-1B genes, splice variants of PTP-1B genes, and PTP-1B gene polymorphisms. 
     By “homologous sequence” is meant, a nucleotide sequence that is shared by one or more polynucleotide sequences, such as genes, gene transcripts and/or non-coding polynucleotides. For example, a homologous sequence can be a nucleotide sequence that is shared by two or more genes encoding related but different proteins, such as different members of a gene family, different protein epitopes, different protein isoforms or completely divergent genes, such as a cytokine and its corresponding receptors. A homologous sequence can be a nucleotide sequence that is shared by two or more non-coding polynucleotides, such as noncoding DNA or RNA, regulatory sequences, introns, and sites of transcriptional control or regulation. Homologous sequences can also include conserved sequence regions shared by more than one polynucleotide sequence. Homology does not need to be perfect homology (e.g., 100%), as partially homologous sequences are also contemplated by the instant invention (e.g., 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80% etc.). 
     By “conserved sequence region” is meant, a nucleotide sequence of one or more regions in a polynucleotide does not vary significantly between generations or from one biological system, subject, or organism to another biological system, subject, or organism. The polynucleotide can include both coding and non-coding DNA and RNA. 
     By “sense region” is meant a nucleotide sequence of a siNA molecule having complementarity to an antisense region of the siNA molecule. In addition, the sense region of a siNA molecule can comprise a nucleic acid sequence having homology with a target nucleic acid sequence. 
     By “antisense region” is meant a nucleotide sequence of a siNA molecule having complementarity to a target nucleic acid sequence. In addition, the antisense region of a siNA molecule can optionally comprise a nucleic acid sequence having complementarity to a sense region of the siNA molecule. 
     By “target nucleic acid” is meant any nucleic acid sequence whose expression or activity is to be modulated. The target nucleic acid can be DNA or RNA. In one embodiment, a target nucleic acid of the invention is PTP-1B RNA or DNA. 
     By “complementarity” is meant that a nucleic acid can form hydrogen bond(s) with another nucleic acid sequence by either traditional Watson-Crick or other non-traditional types. In reference to the nucleic molecules of the present invention, the binding free energy for a nucleic acid molecule with its complementary sequence is sufficient to allow the relevant function of the nucleic acid to proceed, e.g., RNAi activity. Determination of binding free energies for nucleic acid molecules is well known in the art (see, e.g., Turner et al., 1987,  CSH Symp. Quant. Biol. LII pp.  123-133; Frier et al., 1986,  Proc. Nat. Acad. Sci. USA  83:9373-9377; Turner et al., 1987,  J. Am. Chem. Soc.  109:3783-3785). A percent complementarity indicates the percentage of contiguous residues in a nucleic acid molecule that can form hydrogen bonds (e.g., Watson-Crick base pairing) with a second nucleic acid sequence (e.g., 5, 6, 7, 8, 9, or 10 nucleotides out of a total of 10 nucleotides in the first oligonucleotide being based paired to a second nucleic acid sequence having 10 nucleotides represents 50%, 60%, 70%, 80%, 90%, and 100% complementary respectively). “Perfectly complementary” means that all the contiguous residues of a nucleic acid sequence will hydrogen bond with the same number of contiguous residues in a second nucleic acid sequence. In one embodiment, a siNA molecule of the invention comprises about 15 to about 30 or more (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 or more) nucleotides that are complementary to one or more target nucleic acid molecules or a portion thereof. 
     In one embodiment, siNA molecules of the invention that down regulate or reduce PTP-1B gene expression are used for preventing or treating diabetes (e.g., type 1 and type 2), obesity, and/or insulin resistance in a subject or organism. 
     In one embodiment, the siNA molecules of the invention are used to treat diabetes (e.g., type 1 and type 2), obesity, and/or insulin resistance in a subject or organism. 
     In one embodiment of the present invention, each sequence of a siNA molecule of the invention is independently about 15 to about 30 nucleotides in length, in specific embodiments about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length. In another embodiment, the siNA duplexes of the invention independently comprise about 15 to about 30 base pairs (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30). In another embodiment, one or more strands of the siNA molecule of the invention independently comprises about 15 to about 30 nucleotides (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30) that are complementary to a target nucleic acid molecule. In yet another embodiment, siNA molecules of the invention comprising hairpin or circular structures are about 35 to about 55 (e.g., about 35, 40, 45, 50 or 55) nucleotides in length, or about 38 to about 44 (e.g., about 38, 39, 40, 41, 42, 43, or 44) nucleotides in length and comprising about 15 to about 25 (e.g., about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) base pairs. Exemplary siNA molecules of the invention are shown in Table II. Exemplary synthetic siNA molecules of the invention are shown in Table III and/or  FIGS. 4-5 . 
     As used herein “cell” is used in its usual biological sense, and does not refer to an entire multicellular organism, e.g., specifically does not refer to a human. The cell can be present in an organism, e.g., birds, plants and mammals such as humans, cows, sheep, apes, monkeys, swine, dogs, and cats. The cell can be prokaryotic (e.g., bacterial cell) or eukaryotic (e.g., mammalian or plant cell). The cell can be of somatic or germ line origin, totipotent or pluripotent, dividing or non-dividing. The cell can also be derived from or can comprise a gamete or embryo, a stem cell, or a fully differentiated cell. 
     The siNA molecules of the invention are added directly, or can be complexed with cationic lipids, packaged within liposomes, or otherwise delivered to target cells or tissues. The nucleic acid or nucleic acid complexes can be locally administered to relevant tissues ex vivo, or in vivo through local delivery to the lung, with or without their incorporation in biopolymers. In particular embodiments, the nucleic acid molecules of the invention comprise sequences shown in Tables II-III and/or  FIGS. 4-5 . Examples of such nucleic acid molecules consist essentially of sequences defined in these tables and figures. Furthermore, the chemically modified constructs described in Table IV can be applied to any siNA sequence of the invention. 
     In another aspect, the invention provides mammalian cells containing one or more siNA molecules of this invention. The one or more siNA molecules can independently be targeted to the same or different sites. 
     By “RNA” is meant a molecule comprising at least one ribonucleotide residue. By “ribonucleotide” is meant a nucleotide with a hydroxyl group at the 2′ position of a β-D-ribofuranose moiety. The terms include double-stranded RNA, single-stranded RNA, isolated RNA such as partially purified RNA, essentially pure RNA, synthetic RNA, recombinantly produced RNA, as well as altered RNA that differs from naturally occurring RNA by the addition, deletion, substitution and/or alteration of one or more nucleotides. Such alterations can include addition of non-nucleotide material, such as to the end(s) of the siNA or internally, for example at one or more nucleotides of the RNA. Nucleotides in the RNA molecules of the instant invention can also comprise non-standard nucleotides, such as non-naturally occurring nucleotides or chemically synthesized nucleotides or deoxynucleotides. These altered RNAs can be referred to as analogs or analogs of naturally-occurring RNA. 
     By “subject” is meant an organism, which is a donor or recipient of explanted cells or the cells themselves. “Subject” also refers to an organism to which the nucleic acid molecules of the invention can be administered. A subject can be a mammal or mammalian cells, including a human or human cells. 
     The term “phosphorothioate” as used herein refers to an internucleotide linkage having Formula I, wherein Z and/or W comprise a sulfur atom. Hence, the term phosphorothioatc refers to both phosphorothioatc and phosphorodithioatc internucleotide linkages. 
     The term “phosphonoacetate” as used herein refers to an internucleotide linkage having Formula I, wherein Z and/or W comprise an acetyl or protected acetyl group. 
     The term “thiophosphonoacetate” as used herein refers to an internucleotide linkage having Formula I, wherein Z comprises an acetyl or protected acetyl group and W comprises a sulfur atom or alternately W comprises an acetyl or protected acetyl group and Z comprises a sulfur atom. 
     The term “universal base” as used herein refers to nucleotide base analogs that form base pairs with each of the natural DNA/RNA bases with little discrimination between them. Non-limiting examples of universal bases include C-phenyl, C-naphthyl and other aromatic derivatives, inosine, azole carboxamides, and nitroazole derivatives such as 3-nitropyrrole, 4-nitroindole, 5-nitroindole, and 6-nitroindole as known in the art (see for example Loakes, 2001,  Nucleic Acids Research,  29, 2437-2447). 
     The term “acyclic nucleotide” as used herein refers to any nucleotide having an acyclic ribose sugar, for example where any of the ribose carbons (C1, C2, C3, C4, or C5), are independently or in combination absent from the nucleotide. 
     The nucleic acid molecules of the instant invention, individually, or in combination or in conjunction with other drugs, can be used to for preventing or treating diabetes (e.g., type 1 and type 2), obesity, and/or insulin resistance in a subject or organism. 
     In one embodiment, the siNA molecules of the invention can be administered to a subject or can be administered to other appropriate cells (e.g., liver, intestine, pancreas) evident to those skilled in the art, individually or in combination with one or more drugs under conditions suitable for the treatment. 
     In a further embodiment, the siNA molecules can be used in combination with other known treatments to prevent or treat diabetes (e.g., type 1 and type 2), obesity, and/or insulin resistance in a subject or organism. For example, the described molecules could be used in combination with one or more known compounds, treatments, or procedures to prevent or treat diabetes (e.g., type 1 and type 2), obesity, and/or insulin resistance in a subject or organism as are known in the art. 
     In one embodiment, the invention features an expression vector comprising a nucleic acid sequence encoding at least one siNA molecule of the invention, in a manner which allows expression of the siNA molecule. For example, the vector can contain sequence(s) encoding both strands of a siNA molecule comprising a duplex. The vector can also contain sequence(s) encoding a single nucleic acid molecule that is self-complementary and thus forms a siNA molecule. Non-limiting examples of such expression vectors are described in Paul et al., 2002,  Nature Biotechnology,  19, 505; Miyagishi and Taira, 2002,  Nature Biotechnology,  19, 497; Lee et al., 2002,  Nature Biotechnology,  19, 500; and Novina et al., 2002,  Nature Medicine, advance online publication doi: 10.1038/nm725. 
     In another embodiment, the invention features a mammalian cell, for example, a human cell, including an expression vector of the invention. 
     In yet another embodiment, the expression vector of the invention comprises a sequence for a siNA molecule having complementarity to a RNA molecule referred to by a Genbank Accession numbers, for example Genbank Accession Nos. shown in Table I. 
     In one embodiment, an expression vector of the invention comprises a nucleic acid sequence encoding two or more siNA molecules, which can be the same or different. 
     In another aspect of the invention, siNA molecules that interact with target RNA molecules and down-regulate gene encoding target RNA molecules (for example target RNA molecules referred to by Genbank Accession numbers herein) are expressed from transcription units inserted into DNA or RNA vectors. The recombinant vectors can be DNA plasmids or viral vectors. siNA expressing viral vectors can be constructed based on, but not limited to, adeno-associated virus, retrovirus, adenovirus, or alphavirus. The recombinant vectors capable of expressing the siNA molecules can be delivered as described herein, and persist in target cells. Alternatively, viral vectors can be used that provide for transient expression of siNA molecules. Such vectors can be repeatedly administered as necessary. Once expressed, the siNA molecules bind and down-regulate gene function or expression via RNA interference (RNAi). Delivery of siNA expressing vectors can be systemic, such as by intravenous or intramuscular administration, by administration to target cells ex-planted from a subject followed by reintroduction into the subject, or by any other means that would allow for introduction into the desired target cell. 
     By “vectors” is meant any nucleic acid- and/or viral-based technique used to deliver a desired nucleic acid. 
     Other features and advantages of the invention will be apparent from the following description of the preferred embodiments thereof, and from the claims. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  shows a non-limiting example of a scheme for the synthesis of siNA molecules. The complementary siNA sequence strands, strand 1 and strand 2, are synthesized in tandem and are connected by a cleavable linkage, such as a nucleotide succinate or abasic succinate, which can be the same or different from the cleavable linker used for solid phase synthesis on a solid support. The synthesis can be either solid phase or solution phase, in the example shown, the synthesis is a solid phase synthesis. The synthesis is performed such that a protecting group, such as a dimethoxytrityl group, remains intact on the terminal nucleotide of the tandem oligonucleotide. Upon cleavage and deprotection of the oligonucleotide, the two siNA strands spontaneously hybridize to form a siNA duplex, which allows the purification of the duplex by utilizing the properties of the terminal protecting group, for example by applying a trityl on purification method wherein only duplexes/oligonucleotides with the terminal protecting group are isolated. 
         FIG. 2  shows a MALDI-TOF mass spectrum of a purified siNA duplex synthesized by a method of the invention. The two peaks shown correspond to the predicted mass of the separate siNA sequence strands. This result demonstrates that the siNA duplex generated from tandem synthesis can be purified as a single entity using a simple trityl-on purification methodology. 
         FIG. 3  shows a non-limiting proposed mechanistic representation of target RNA degradation involved in RNAi. Double-stranded RNA (dsRNA), which is generated by RNA-dependent RNA polymerase (RdRP) from foreign single-stranded RNA, for example viral, transposon, or other exogenous RNA, activates the DICER enzyme that in turn generates siNA duplexes. Alternately, synthetic or expressed siNA can be introduced directly into a cell by appropriate means. An active siNA complex forms which recognizes a target RNA, resulting in degradation of the target RNA by the RISC endonuclease complex or in the synthesis of additional RNA by RNA-dependent RNA polymerase (RdRP), which can activate DICER and result in additional siNA molecules, thereby amplifying the RNAi response. 
         FIG. 4A-F  shows non-limiting examples of chemically-modified siNA constructs of the present invention. In the figure, N stands for any nucleotide (adenosine, guanosine, cytosine, uridine, or optionally thymidine, for example thymidine can be substituted in the overhanging regions designated by parenthesis (N N). Various modifications are shown for the sense and antisense strands of the siNA constructs. 
         FIG. 4A : The sense strand comprises 21 nucleotides wherein the two terminal 3′-nucleotides are optionally base paired and wherein all nucleotides present are ribonucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. The antisense strand comprises 21 nucleotides, optionally having a 3′-terminal glyceryl moiety wherein the two terminal 3′-nucleotides are optionally complementary to the target RNA sequence, and wherein all nucleotides present are ribonucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. A modified internucleotide linkage, such as a phosphorothioate, phosphorodithioate or other modified internucleotide linkage as described herein, shown as “s”, optionally connects the (N N) nucleotides in the antisense strand. 
         FIG. 4B : The sense strand comprises 21 nucleotides wherein the two terminal 3′-nucleotides are optionally base paired and wherein all pyrimidine nucleotides that may be present are 2′deoxy-2′-fluoro modified nucleotides and all purine nucleotides that may be present are 2′-O-methyl modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. The antisense strand comprises 21 nucleotides, optionally having a 3′-terminal glyceryl moiety and wherein the two terminal 3′-nucleotides are optionally complementary to the target RNA sequence, and wherein all pyrimidine nucleotides that may be present are 2′-deoxy-2′-fluoro modified nucleotides and all purine nucleotides that may be present are 2′-O-methyl modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. A modified internucleotide linkage, such as a phosphorothioate, phosphorodithioate or other modified internucleotide linkage as described herein, shown as “s”, optionally connects the (N N) nucleotides in the sense and antisense strand. 
         FIG. 4C : The sense strand comprises 21 nucleotides having 5′- and 3′-terminal cap moieties wherein the two terminal 3′-nucleotides are optionally base paired and wherein all pyrimidine nucleotides that may be present are 2′-O-methyl or 2′-deoxy-2′-fluoro modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. The antisense strand comprises 21 nucleotides, optionally having a 3′-terminal glyceryl moiety and wherein the two terminal 3′-nucleotides are optionally complementary to the target RNA sequence, and wherein all pyrimidine nucleotides that may be present are 2′-deoxy-2′-fluoro modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. A modified internucleotide linkage, such as a phosphorothioate, phosphorodithioate or other modified internucleotide linkage as described herein, shown as “s”, optionally connects the (N N) nucleotides in the antisense strand. 
         FIG. 4D : The sense strand comprises 21 nucleotides having 5′- and 3′-terminal cap moieties wherein the two terminal 3′-nucleotides are optionally base paired and wherein all pyrimidine nucleotides that may be present are 2′-deoxy-2′-fluoro modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein and wherein and all purine nucleotides that may be present are 2′-deoxy nucleotides. The antisense strand comprises 21 nucleotides, optionally having a 3′-terminal glyceryl moiety and wherein the two terminal 3′-nucleotides are optionally complementary to the target RNA sequence, wherein all pyrimidine nucleotides that may be present are 2′-deoxy-2′-fluoro modified nucleotides and all purine nucleotides that may be present are 2′-O-methyl modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. A modified internucleotide linkage, such as a phosphorothioate, phosphorodithioate or other modified internucleotide linkage as described herein, shown as “s”, optionally connects the (N N) nucleotides in the antisense strand. 
         FIG. 4E : The sense strand comprises 21 nucleotides having 5′- and 3′-terminal cap moieties wherein the two terminal 3′-nucleotides are optionally base paired and wherein all pyrimidine nucleotides that may be present are 2′-deoxy-2′-fluoro modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. The antisense strand comprises 21 nucleotides, optionally having a 3′-terminal glyceryl moiety and wherein the two terminal 3′-nucleotides are optionally complementary to the target RNA sequence, and wherein all pyrimidine nucleotides that may be present are 2′-deoxy-2′-fluoro modified nucleotides and all purine nucleotides that may be present are 2′-O-methyl modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. A modified internucleotide linkage, such as a phosphorothioate, phosphorodithioate or other modified internucleotide linkage as described herein, shown as “s”, optionally connects the (N N) nucleotides in the antisense strand. 
         FIG. 4F : The sense strand comprises 21 nucleotides having 5′- and 3′-terminal cap moieties wherein the two terminal 3′-nucleotides are optionally base paired and wherein all pyrimidine nucleotides that may be present are 2′-deoxy-2′-fluoro modified nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein and wherein and all purine nucleotides that may be present are 2′-deoxy nucleotides. The antisense strand comprises 21 nucleotides, optionally having a 3′-terminal glyceryl moiety and wherein the two terminal 3′-nucleotides are optionally complementary to the target RNA sequence, and having one 3′-terminal phosphorothioate internucleotide linkage and wherein all pyrimidine nucleotides that may be present are 2′-deoxy-2′-fluoro modified nucleotides and all purine nucleotides that may be present are 2′-deoxy nucleotides except for (N N) nucleotides, which can comprise ribonucleotides, deoxynucleotides, universal bases, or other chemical modifications described herein. A modified internucleotide linkage, such as a phosphorothioate, phosphorodithioate or other modified internucleotide linkage as described herein, shown as “s”, optionally connects the (N N) nucleotides in the antisense strand. The antisense strand of constructs A-F comprise sequence complementary to any target nucleic acid sequence of the invention. Furthermore, when a glyceryl moiety (L) is present at the 3′-end of the antisense strand for any construct shown in  FIG. 4  A-F, the modified internucleotide linkage is optional. 
         FIG. 5A-F  shows non-limiting examples of specific chemically-modified siNA sequences of the invention. A-F applies the chemical modifications described in  FIG. 4A-F  to a PTP-1B siNA sequence. Such chemical modifications can be applied to any PTP-1B sequence. 
         FIG. 6  shows non-limiting examples of different siNA constructs of the invention. The examples shown (constructs 1, 2, and 3) have 19 representative base pairs; however, different embodiments of the invention include any number of base pairs described herein. Bracketed regions represent nucleotide overhangs, for example, comprising about 1, 2, 3, or 4 nucleotides in length, preferably about 2 nucleotides. Constructs 1 and 2 can be used independently for RNAi activity. Construct 2 can comprise a polynucleotide or non-nucleotide linker, which can optionally be designed as a biodegradable linker. In one embodiment, the loop structure shown in construct 2 can comprise a biodegradable linker that results in the formation of construct 1 in vivo and/or in vitro. In another example, construct 3 can be used to generate construct 2 under the same principle wherein a linker is used to generate the active siNA construct 2 in vivo and/or in vitro, which can optionally utilize another biodegradable linker to generate the active siNA construct 1 in vivo and/or in vitro. As such, the stability and/or activity of the siNA constructs can be modulated based on the design of the siNA construct for use in vivo or in vitro and/or in vitro. 
         FIG. 7A-C  is a diagrammatic representation of a scheme utilized in generating an expression cassette to generate siNA hairpin constructs. 
         FIG. 7A : A DNA oligomer is synthesized with a 5′-restriction site (R1) sequence followed by a region having sequence identical (sense region of siNA) to a predetermined PTP-1B target sequence, wherein the sense region comprises, for example, about 19, 20, 21, or 22 nucleotides (N) in length, which is followed by a loop sequence of defined sequence (X), comprising, for example, about 3 to about 10 nucleotides. 
         FIG. 7B : The synthetic construct is then extended by DNA polymerase to generate a hairpin structure having self-complementary sequence that will result in a siNA transcript having specificity for a PTP-1B target sequence and having self-complementary sense and antisense regions. 
         FIG. 7C : The construct is heated (for example to about 95° C.) to linearize the sequence, thus allowing extension of a complementary second DNA strand using a primer to the 3′-restriction sequence of the first strand. The double-stranded DNA is then inserted into an appropriate vector for expression in cells. The construct can be designed such that a 3′-terminal nucleotide overhang results from the transcription, for example, by engineering restriction sites and/or utilizing a poly-U termination region as described in Paul et al., 2002,  Nature Biotechnology,  29, 505-508. 
         FIG. 8A-C  is a diagrammatic representation of a scheme utilized in generating an expression cassette to generate double-stranded siNA constructs. 
         FIG. 8A : A DNA oligomer is synthesized with a 5′-restriction (R1) site sequence followed by a region having sequence identical (sense region of siNA) to a predetermined PTP-1B target sequence, wherein the sense region comprises, for example, about 19, 20, 21, or 22 nucleotides (N) in length, and which is followed by a 3′-restriction site (R2) which is adjacent to a loop sequence of defined sequence (X). 
         FIG. 8B : The synthetic construct is then extended by DNA polymerase to generate a hairpin structure having self-complementary sequence. 
         FIG. 8C : The construct is processed by restriction enzymes specific to R1 and R2 to generate a double-stranded DNA which is then inserted into an appropriate vector for expression in cells. The transcription cassette is designed such that a U6 promoter region flanks each side of the dsDNA which generates the separate sense and antisense strands of the siNA. Poly T termination sequences can be added to the constructs to generate U overhangs in the resulting transcript. 
         FIG. 9A-E  is a diagrammatic representation of a method used to determine target sites for siNA mediated RNAi within a particular target nucleic acid sequence, such as messenger RNA. 
         FIG. 9A : A pool of siNA oligonucleotides are synthesized wherein the antisense region of the siNA constructs has complementarity to target sites across the target nucleic acid sequence, and wherein the sense region comprises sequence complementary to the antisense region of the siNA. 
         FIGS. 9B&amp;C : ( FIG. 9B ) The sequences are pooled and are inserted into vectors such that ( FIG. 9C ) transfection of a vector into cells results in the expression of the siNA. 
         FIG. 9D : Cells are sorted based on phenotypic change that is associated with modulation of the target nucleic acid sequence. 
         FIG. 9E : The siNA is isolated from the sorted cells and is sequenced to identify efficacious target sites within the target nucleic acid sequence. 
         FIG. 10  shows non-limiting examples of different stabilization chemistries (1-10) that can be used, for example, to stabilize the 3′-end of siNA sequences of the invention, including (1) [3-3′]-inverted deoxyribose; (2) deoxyribonucleotide; (3) [5′-3′]-3′-deoxyribonucleotide; (4) [5′-3′]-ribonucleotide; (5) [5′-3′]-3′-O-methyl ribonucleotide; (6) 3′-glyceryl; (7) [3′-5′]-3′-deoxyribonucleotide; (8) [3′-3′]-deoxyribonucleotide; (9) [5′-2′]-deoxyribonucleotide; and (10) [5-3′]-dideoxyribonucleotide. In addition to modified and unmodified backbone chemistries indicated in the figure, these chemistries can be combined with different backbone modifications as described herein, for example, backbone modifications having Formula I. In addition, the 2′-deoxy nucleotide shown 5′ to the terminal modifications shown can be another modified or unmodified nucleotide or non-nucleotide described herein, for example modifications having any of Formulae I-VII or any combination thereof. 
         FIG. 11  shows a non-limiting example of a strategy used to identify chemically modified siNA constructs of the invention that are nuclease resistance while preserving the ability to mediate RNAi activity. Chemical modifications are introduced into the siNA construct based on educated design parameters (e.g. introducing 2′-modifications, base modifications, backbone modifications, terminal cap modifications etc). The modified construct in tested in an appropriate system (e.g. human serum for nuclease resistance, shown, or an animal model for PK/delivery parameters). In parallel, the siNA construct is tested for RNAi activity, for example in a cell culture system such as a luciferase reporter assay). Lead siNA constructs are then identified which possess a particular characteristic while maintaining RNAi activity, and can be further modified and assayed once again. This same approach can be used to identify siNA-conjugate molecules with improved pharmacokinetic profiles, delivery, and RNAi activity. 
         FIG. 12  shows non-limiting examples of phosphorylated siNA molecules of the invention, including linear and duplex constructs and asymmetric derivatives thereof. 
         FIG. 13  shows non-limiting examples of chemically modified terminal phosphate groups of the invention. 
         FIG. 14A  shows a non-limiting example of methodology used to design self complementary DFO constructs utilizing palindrome and/or repeat nucleic acid sequences that are identified in a target nucleic acid sequence. (i) A palindrome or repeat sequence is identified in a nucleic acid target sequence. (ii) A sequence is designed that is complementary to the target nucleic acid sequence and the palindrome sequence. (iii) An inverse repeat sequence of the non-palindrome/repeat portion of the complementary sequence is appended to the 3′-end of the complementary sequence to generate a self complementary DFO molecule comprising sequence complementary to the nucleic acid target. (iv) The DFO molecule can self-assemble to form a double stranded oligonucleotide.  FIG. 14B  shows a non-limiting representative example of a duplex forming oligonucleotide sequence.  FIG. 14C  shows a non-limiting example of the self assembly schematic of a representative duplex forming oligonucleotide sequence.  FIG. 14D  shows a non-limiting example of the self assembly schematic of a representative duplex forming oligonucleotide sequence followed by interaction with a target nucleic acid sequence resulting in modulation of gene expression. 
         FIG. 15  shows a non-limiting example of the design of self complementary DFO constructs utilizing palindrome and/or repeat nucleic acid sequences that are incorporated into the DFO constructs that have sequence complementary to any target nucleic acid sequence of interest. Incorporation of these palindrome/repeat sequences allow the design of DFO constructs that form duplexes in which each strand is capable of mediating modulation of target gene expression, for example by RNAi. First, the target sequence is identified. A complementary sequence is then generated in which nucleotide or non-nucleotide modifications (shown as X or Y) are introduced into the complementary sequence that generate an artificial palindrome (shown as XYXYXY in the Figure). An inverse repeat of the non-palindrome/repeat complementary sequence is appended to the 3′-end of the complementary sequence to generate a self complementary DFO comprising sequence complementary to the nucleic acid target. The DFO can self-assemble to form a double stranded oligonucleotide. 
         FIG. 16  shows non-limiting examples of multifunctional siNA molecules of the invention comprising two separate polynucleotide sequences that are each capable of mediating RNAi directed cleavage of differing target nucleic acid sequences.  FIG. 16A  shows a non-limiting example of a multifunctional siNA molecule having a first region that is complementary to a first target nucleic acid sequence (complementary region 1) and a second region that is complementary to a second target nucleic acid sequence (complementary region 2), wherein the first and second complementary regions are situated at the 3′-ends of each polynucleotide sequence in the multifunctional siNA. The dashed portions of each polynucleotide sequence of the multifunctional siNA construct have complementarity with regard to corresponding portions of the siNA duplex, but do not have complementarity to the target nucleic acid sequences.  FIG. 16B  shows a non-limiting example of a multifunctional siNA molecule having a first region that is complementary to a first target nucleic acid sequence (complementary region 1) and a second region that is complementary to a second target nucleic acid sequence (complementary region 2), wherein the first and second complementary regions are situated at the 5′-ends of each polynucleotide sequence in the multifunctional siNA. The dashed portions of each polynucleotide sequence of the multifunctional siNA construct have complementarity with regard to corresponding portions of the siNA duplex, but do not have complementarity to the target nucleic acid sequences. 
         FIG. 17  shows non-limiting examples of multifunctional siNA molecules of the invention comprising a single polynucleotide sequence comprising distinct regions that are each capable of mediating RNAi directed cleavage of differing target nucleic acid sequences.  FIG. 17A  shows a non-limiting example of a multifunctional siNA molecule having a first region that is complementary to a first target nucleic acid sequence (complementary region 1) and a second region that is complementary to a second target nucleic acid sequence (complementary region 2), wherein the second complementary region is situated at the 3′-end of the polynucleotide sequence in the multifunctional siNA. The dashed portions of each polynucleotide sequence of the multifunctional siNA construct have complementarity with regard to corresponding portions of the siNA duplex, but do not have complementarity to the target nucleic acid sequences.  FIG. 17B  shows a non-limiting example of a multifunctional siNA molecule having a first region that is complementary to a first target nucleic acid sequence (complementary region 1) and a second region that is complementary to a second target nucleic acid sequence (complementary region 2), wherein the first complementary region is situated at the 5′-end of the polynucleotide sequence in the multifunctional siNA. The dashed portions of each polynucleotide sequence of the multifunctional siNA construct have complementarity with regard to corresponding portions of the siNA duplex, but do not have complementarity to the target nucleic acid sequences. In one embodiment, these multifunctional siNA constructs are processed in vivo or in vitro to generate multifunctional siNA constructs as shown in  FIG. 16 . 
         FIG. 18  shows non-limiting examples of multifunctional siNA molecules of the invention comprising two separate polynucleotide sequences that are each capable of mediating RNAi directed cleavage of differing target nucleic acid sequences and wherein the multifunctional siNA construct further comprises a self complementary, palindrome, or repeat region, thus enabling shorter bifunctional siNA constructs that can mediate RNA interference against differing target nucleic acid sequences.  FIG. 18A  shows a non-limiting example of a multifunctional siNA molecule having a first region that is complementary to a first target nucleic acid sequence (complementary region 1) and a second region that is complementary to a second target nucleic acid sequence (complementary region 2), wherein the first and second complementary regions are situated at the 3′-ends of each polynucleotide sequence in the multifunctional siNA, and wherein the first and second complementary regions further comprise a self complementary, palindrome, or repeat region. The dashed portions of each polynucleotide sequence of the multifunctional siNA construct have complementarity with regard to corresponding portions of the siNA duplex, but do not have complementarity to the target nucleic acid sequences.  FIG. 18B  shows a non-limiting example of a multifunctional siNA molecule having a first region that is complementary to a first target nucleic acid sequence (complementary region 1) and a second region that is complementary to a second target nucleic acid sequence (complementary region 2), wherein the first and second complementary regions are situated at the 5′-ends of each polynucleotide sequence in the multifunctional siNA, and wherein the first and second complementary regions further comprise a self complementary, palindrome, or repeat region. The dashed portions of each polynucleotide sequence of the multifunctional siNA construct have complementarity with regard to corresponding portions of the siNA duplex, but do not have complementarity to the target nucleic acid sequences. 
         FIG. 19  shows non-limiting examples of multifunctional siNA molecules of the invention comprising a single polynucleotide sequence comprising distinct regions that are each capable of mediating RNAi directed cleavage of differing target nucleic acid sequences and wherein the multifunctional siNA construct further comprises a self complementary, palindrome, or repeat region, thus enabling shorter bifunctional siNA constructs that can mediate RNA interference against differing target nucleic acid sequences.  FIG. 19A  shows a non-limiting example of a multifunctional siNA molecule having a first region that is complementary to a first target nucleic acid sequence (complementary region 1) and a second region that is complementary to a second target nucleic acid sequence (complementary region 2), wherein the second complementary region is situated at the 3′-end of the polynucleotide sequence in the multifunctional siNA, and wherein the first and second complementary regions further comprise a self complementary, palindrome, or repeat region. The dashed portions of each polynucleotide sequence of the multifunctional siNA construct have complementarity with regard to corresponding portions of the siNA duplex, but do not have complementarity to the target nucleic acid sequences.  FIG. 19B  shows a non-limiting example of a multifunctional siNA molecule having a first region that is complementary to a first target nucleic acid sequence (complementary region 1) and a second region that is complementary to a second target nucleic acid sequence (complementary region 2), wherein the first complementary region is situated at the 5′-end of the polynucleotide sequence in the multifunctional siNA, and wherein the first and second complementary regions further comprise a self complementary, palindrome, or repeat region. The dashed portions of each polynucleotide sequence of the multifunctional siNA construct have complementarity with regard to corresponding portions of the siNA duplex, but do not have complementarity to the target nucleic acid sequences. In one embodiment, these multifunctional siNA constructs are processed in vivo or in vitro to generate multifunctional siNA constructs as shown in  FIG. 18 . 
         FIG. 20  shows a non-limiting example of how multifunctional siNA molecules of the invention can target two separate target nucleic acid molecules, such as separate RNA molecules encoding differing proteins, for example, a cytokine and its corresponding receptor, differing viral strains, a virus and a cellular protein involved in viral infection or replication, or differing proteins involved in a common or divergent biologic pathway that is implicated in the maintenance of progression of disease. Each strand of the multifunctional siNA construct comprises a region having complementarity to separate target nucleic acid molecules. The multifunctional siNA molecule is designed such that each strand of the siNA can be utilized by the RISC complex to initiate RNA interference mediated cleavage of its corresponding target. These design parameters can include destabilization of each end of the siNA construct (see for example Schwarz et al., 2003,  Cell,  115, 199-208). Such destabilization can be accomplished for example by using guanosine-cytidine base pairs, alternate base pairs (e.g., wobbles), or destabilizing chemically modified nucleotides at terminal nucleotide positions as is known in the art. 
         FIG. 21  shows a non-limiting example of how multifunctional siNA molecules of the invention can target two separate target nucleic acid sequences within the same target nucleic acid molecule, such as alternate coding regions of a RNA, coding and non-coding regions of a RNA, or alternate splice variant regions of a RNA. Each strand of the multifunctional siNA construct comprises a region having complementarity to the separate regions of the target nucleic acid molecule. The multifunctional siNA molecule is designed such that each strand of the siNA can be utilized by the RISC complex to initiate RNA interference mediated cleavage of its corresponding target region. These design parameters can include destabilization of each end of the siNA construct (see for example Schwarz et al., 2003,  Cell,  115, 199-208). Such destabilization can be accomplished for example by using guanosine-cytidine base pairs, alternate base pairs (e.g., wobbles), or destabilizing chemically modified nucleotides at terminal nucleotide positions as is known in the art. 
         FIG. 22(A-H)  shows non-limiting examples of tethered multifunctional siNA constructs of the invention. In the examples shown, a linker (e.g., nucleotide or non-nucleotide linker) connects two siNA regions (e.g., two sense, two antisense, or alternately a sense and an antisense region together. Separate sense (or sense and antisense) sequences corresponding to a first target sequence and second target sequence are hybridized to their corresponding sense and/or antisense sequences in the multifunctional siNA. In addition, various conjugates, ligands, aptamers, polymers or reporter molecules can be attached to the linker region for selective or improved delivery and/or pharmacokinetic properties. 
         FIG. 23  shows a non-limiting example of various dendrimer based multifunctional siNA designs. 
         FIG. 24  shows a non-limiting example of various supramolecular multifunctional siNA designs. 
         FIG. 25  shows a non-limiting example of a dicer enabled multifunctional siNA design using a 30 nucleotide precursor siNA construct. A 30 base pair duplex is cleaved by Dicer into 22 and 8 base pair products from either end (8 b.p. fragments not shown). For ease of presentation the overhangs generated by dicer are not shown—but can be compensated for. Three targeting sequences are shown. The required sequence identity overlapped is indicated by grey boxes. The N&#39;s of the parent 30 b.p. siNA are suggested sites of 2′-OH positions to enable Dicer cleavage if this is tested in stabilized chemistries. Note that processing of a 30 mer duplex by Dicer RNase III does not give a precise 22+8 cleavage, but rather produces a series of closely related products (with 22+8 being the primary site). Therefore, processing by Dicer will yield a series of active siNAs. 
         FIG. 26  shows a non-limiting example of a dicer enabled multifunctional siNA design using a 40 nucleotide precursor siNA construct. A 40 base pair duplex is cleaved by Dicer into 20 base pair products from either end. For ease of presentation the overhangs generated by dicer are not shown—but can be compensated for. Four targeting sequences are shown. The target sequences having homology are enclosed by boxes. This design format can be extended to larger RNAs. If chemically stabilized siNAs are bound by Dicer, then strategically located ribonucleotide linkages can enable designer cleavage products that permit our more extensive repertoire of multifunctional designs. For example cleavage products not limited to the Dicer standard of approximately 22-nucleotides can allow multifunctional siNA constructs with a target sequence identity overlap ranging from, for example, about 3 to about 15 nucleotides. 
         FIG. 27  shows a non-limiting example of additional multifunctional siNA construct designs of the invention. In one example, a conjugate, ligand, aptamer, label, or other moiety is attached to a region of the multifunctional siNA to enable improved delivery or pharmacokinetic profiling. 
         FIG. 28  shows a non-limiting example of additional multifunctional siNA construct designs of the invention. In one example, a conjugate, ligand, aptamer, label, or other moiety is attached to a region of the multifunctional siNA to enable improved delivery or pharmacokinetic profiling. 
         FIG. 29  shows a non-limiting example of reduction of PTP-1B mRNA in A549 cells mediated by chemically-modified siNAs that target PTP-1B mRNA. A549 cells were transfected with 0.25 ug/well of lipid complexed with 25 nM siNA. A siNA construct comprising ribonucleotides and 3′-terminal dithymidine caps (compound numbers 31018/31094) was compared to a chemically modified siNA construct comprising 2′-deoxy-2′-fluoro pyrimidine nucleotides and purine ribonucleotides in which the sense strand of the siNA is further modified with 5′ and 3′-terminal inverted deoxyabasic caps and the antisense strand comprises a 3′-terminal phosphorothioate internucleotide linkage (compound number 31306/31307), which was also compared to a matched chemistry inverted control (compound number 31318/31319). In addition, the siNA constructs were also compared to untreated cells, cells transfected with lipid and scrambled siNA constructs (Scram1 and Scram2), and cells transfected with lipid alone (transfection control). As shown in the figure, both siNA constructs show significant reduction of PTP-1B RNA expression compared to appropriate controls. 
         FIG. 30  shows a non-limiting example of a dose response study of reduction of PTP-1B mRNA in Hepal-6 cells mediated by chemically-modified siNAs that target site 761 PTP-1B mRNA. Dose response curves were carried out using PTP-1B site 761 stab 4/8 and 7/8 constructs (Table III) at an initial concentration of 50 nM diluted two-fold down to 781 pM. After a 24 hour incubation, the cells were harvested, total RNA isolated, and the level of PTP-1B mRNA was determined by real time RT_PCR using PTP-1B specific primers and probe. 
         FIG. 31  shows a non-limiting example of a cholesterol linked phosphoramidite that can be used to synthesize cholesterol conjugated siNA molecules of the invention. An example is shown with the cholesterol moiety linked to the 5′-end of the sense strand of a siNA molecule. 
         FIG. 32  shows a non-limiting example of reduction of PTP-1B mRNA in CD-1 mice following systemic intravenous administration of cholesterol conjugated siNA molecules targeting PTP-1B RNA compared to a matched chemistry cholesterol conjugated irrelevant control and a PBS control (no siNA). Mice, (N=5 per group) were dosed iv 30 mg/kg/BID for three days. Sixteen hours post the last dose, animals were sacrificed. Total RNA was isolated from liver, and RT-PCR was performed with PTP-1B specific primers and probe. As shown in the figure, active cholesterol conjugated siNA molecules provided between 67% and 72% inhibition of PTP-1B RNA. 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     Mechanism of Action of Nucleic Acid Molecules of the Invention 
     The discussion that follows discusses the proposed mechanism of RNA interference mediated by short interfering RNA as is presently known, and is not meant to be limiting and is not an admission of prior art. Applicant demonstrates herein that chemically-modified short interfering nucleic acids possess similar or improved capacity to mediate RNAi as do siRNA molecules and are expected to possess improved stability and activity in vivo; therefore, this discussion is not meant to be limiting only to siRNA and can be applied to siNA as a whole. By “improved capacity to mediate RNAi” or “improved RNAi activity” is meant to include RNAi activity measured in vitro and/or in vivo where the RNAi activity is a reflection of both the ability of the siNA to mediate RNAi and the stability of the siNAs of the invention. In this invention, the product of these activities can be increased in vitro and/or in vivo compared to an all RNA siRNA or a siNA containing a plurality of ribonucleotides. In some cases, the activity or stability of the siNA molecule can be decreased (i.e., less than ten-fold), but the overall activity of the siNA molecule is enhanced in vitro and/or in vivo. 
     RNA interference refers to the process of sequence specific post-transcriptional gene silencing in animals mediated by short interfering RNAs (siRNAs) (Fire et al., 1998,  Nature,  391, 806). The corresponding process in plants is commonly referred to as post-transcriptional gene silencing or RNA silencing and is also referred to as quelling in fungi. The process of post-transcriptional gene silencing is thought to be an evolutionarily-conserved cellular defense mechanism used to prevent the expression of foreign genes which is commonly shared by diverse flora and phyla (Fire et al., 1999,  Trends Genet.,  15, 358). Such protection from foreign gene expression may have evolved in response to the production of double-stranded RNAs (dsRNAs) derived from viral infection or the random integration of transposon elements into a host genome via a cellular response that specifically destroys homologous single-stranded RNA or viral genomic RNA. The presence of dsRNA in cells triggers the RNAi response though a mechanism that has yet to be fully characterized. This mechanism appears to be different from the interferon response that results from dsRNA-mediated activation of protein kinase PKR and 2′,5′-oligoadenylate synthetase resulting in non-specific cleavage of mRNA by ribonuclease L. 
     The presence of long dsRNAs in cells stimulates the activity of a ribonuclease III enzyme referred to as Dicer. Dicer is involved in the processing of the dsRNA into short pieces of dsRNA known as short interfering RNAs (siRNAs) (Berstein et al., 2001,  Nature,  409, 363). Short interfering RNAs derived from Dicer activity are typically about 21 to about 23 nucleotides in length and comprise about 19 base pair duplexes. Dicer has also been implicated in the excision of 21- and 22-nucleotide small temporal RNAs (stRNAs) from precursor RNA of conserved structure that are implicated in translational control (Hutvagner et al., 2001,  Science,  293, 834). The RNAi response also features an endonuclease complex containing a siRNA, commonly referred to as an RNA-induced silencing complex (RISC), which mediates cleavage of single-stranded RNA having sequence homologous to the siRNA. Cleavage of the target RNA takes place in the middle of the region complementary to the guide sequence of the siRNA duplex (Elbashir et al., 2001,  Genes Dev.,  15, 188). In addition, RNA interference can also involve small RNA (e.g., micro-RNA or miRNA) mediated gene silencing, presumably though cellular mechanisms that regulate chromatin structure and thereby prevent transcription of target gene sequences (see for example Allshire, 2002,  Science,  297, 1818-1819; Volpe et al., 2002,  Science,  297, 1833-1837; Jenuwein, 2002,  Science,  297, 2215-2218; and Hall et al., 2002,  Science,  297, 2232-2237). As such, siNA molecules of the invention can be used to mediate gene silencing via interaction with RNA transcripts or alternately by interaction with particular gene sequences, wherein such interaction results in gene silencing either at the transcriptional level or post-transcriptional level. 
     RNAi has been studied in a variety of systems. Fire et al., 1998,  Nature,  391, 806, were the first to observe RNAi in  C. elegans . Wianny and Goetz, 1999,  Nature Cell Biol.,  2, 70, describe RNAi mediated by dsRNA in mouse embryos. Hammond et al., 2000,  Nature,  404, 293, describe RNAi in  Drosophila  cells transfected with dsRNA. Elbashir et al., 2001,  Nature,  411, 494, describe RNAi induced by introduction of duplexes of synthetic 21-nucleotide RNAs in cultured mammalian cells including human embryonic kidney and HeLa cells. Recent work in  Drosophila  embryonic lysates has revealed certain requirements for siRNA length, structure, chemical composition, and sequence that are essential to mediate efficient RNAi activity. These studies have shown that 21 nucleotide siRNA duplexes are most active when containing two 2-nucleotide 3′-terminal nucleotide overhangs. Furthermore, substitution of one or both siRNA strands with 2′-deoxy or 2′-O-methyl nucleotides abolishes RNAi activity, whereas substitution of 3′-terminal siRNA nucleotides with deoxy nucleotides was shown to be tolerated. Mismatch sequences in the center of the siRNA duplex were also shown to abolish RNAi activity. In addition, these studies also indicate that the position of the cleavage site in the target RNA is defined by the 5′-end of the siRNA guide sequence rather than the 3′-end (Elbashir et al., 2001,  EMBO J.,  20, 6877). Other studies have indicated that a 5′-phosphate on the target-complementary strand of a siRNA duplex is required for siRNA activity and that ATP is utilized to maintain the 5′-phosphate moiety on the siRNA (Nykanen et al., 2001,  Cell,  107, 309); however, siRNA molecules lacking a 5′-phosphate are active when introduced exogenously, suggesting that 5′-phosphorylation of siRNA constructs may occur in vivo. 
     Duplex Forming Oligonucleotides (DFO) of the Invention 
     In one embodiment, the invention features siNA molecules comprising duplex forming oligonucleotides (DFO) that can self-assemble into double stranded oligonucleotides. The duplex forming oligonucleotides of the invention can be chemically synthesized or expressed from transcription units and/or vectors. The DFO molecules of the instant invention provide useful reagents and methods for a variety of therapeutic, diagnostic, agricultural, veterinary, target validation, genomic discovery, genetic engineering and pharmacogenomic applications. 
     Applicant demonstrates herein that certain oligonucleotides, referred to herein for convenience but not limitation as duplex forming oligonucleotides or DFO molecules, are potent mediators of sequence specific regulation of gene expression. The oligonucleotides of the invention are distinct from other nucleic acid sequences known in the art (e.g., siRNA, miRNA, stRNA, shRNA, antisense oligonucleotides etc.) in that they represent a class of linear polynucleotide sequences that are designed to self-assemble into double stranded oligonucleotides, where each strand in the double stranded oligonucleotides comprises a nucleotide sequence that is complementary to a target nucleic acid molecule. Nucleic acid molecules of the invention can thus self assemble into functional duplexes in which each strand of the duplex comprises the same polynucleotide sequence and each strand comprises a nucleotide sequence that is complementary to a target nucleic acid molecule. 
     Generally, double stranded oligonucleotides are formed by the assembly of two distinct oligonucleotide sequences where the oligonucleotide sequence of one strand is complementary to the oligonucleotide sequence of the second strand; such double stranded oligonucleotides are assembled from two separate oligonucleotides, or from a single molecule that folds on itself to form a double stranded structure, often referred to in the field as hairpin stem-loop structure (e.g., shRNA or short hairpin RNA). These double stranded oligonucleotides known in the art all have a common feature in that each strand of the duplex has a distinct nucleotide sequence. 
     Distinct from the double stranded nucleic acid molecules known in the art, the applicants have developed a novel, potentially cost effective and simplified method of forming a double stranded nucleic acid molecule starting from a single stranded or linear oligonucleotide. The two strands of the double stranded oligonucleotide formed according to the instant invention have the same nucleotide sequence and are not covalently linked to each other. Such double-stranded oligonucleotides molecules can be readily linked post-synthetically by methods and reagents known in the art and are within the scope of the invention. In one embodiment, the single stranded oligonucleotide of the invention (the duplex forming oligonucleotide) that forms a double stranded oligonucleotide comprises a first region and a second region, where the second region includes a nucleotide sequence that is an inverted repeat of the nucleotide sequence in the first region, or a portion thereof, such that the single stranded oligonucleotide self assembles to form a duplex oligonucleotide in which the nucleotide sequence of one strand of the duplex is the same as the nucleotide sequence of the second strand. Non-limiting examples of such duplex forming oligonucleotides are illustrated in  FIGS. 14 and 15 . These duplex forming oligonucleotides (DFOs) can optionally include certain palindrome or repeat sequences where such palindrome or repeat sequences are present in between the first region and the second region of the DFO. 
     In one embodiment, the invention features a duplex forming oligonucleotide (DFO) molecule, wherein the DFO comprises a duplex forming self complementary nucleic acid sequence that has nucleotide sequence complementary to a PTP-1B target nucleic acid sequence. The DFO molecule can comprise a single self complementary sequence or a duplex resulting from assembly of such self complementary sequences. 
     In one embodiment, a duplex forming oligonucleotide (DFO) of the invention comprises a first region and a second region, wherein the second region comprises a nucleotide sequence comprising an inverted repeat of nucleotide sequence of the first region such that the DFO molecule can assemble into a double stranded oligonucleotide. Such double stranded oligonucleotides can act as a short interfering nucleic acid (siNA) to modulate gene expression. Each strand of the double stranded oligonucleotide duplex formed by DFO molecules of the invention can comprise a nucleotide sequence region that is complementary to the same nucleotide sequence in a target nucleic acid molecule (e.g., target PTP-1B RNA). 
     In one embodiment, the invention features a single stranded DFO that can assemble into a double stranded oligonucleotide. The applicant has surprisingly found that a single stranded oligonucleotide with nucleotide regions of self complementarity can readily assemble into duplex oligonucleotide constructs. Such DFOs can assemble into duplexes that can inhibit gene expression in a sequence specific manner. The DFO molecules of the invention comprise a first region with nucleotide sequence that is complementary to the nucleotide sequence of a second region and where the sequence of the first region is complementary to a target nucleic acid (e.g., RNA). The DFO can form a double stranded oligonucleotide wherein a portion of each strand of the double stranded oligonucleotide comprises a sequence complementary to a target nucleic acid sequence. 
     In one embodiment, the invention features a double stranded oligonucleotide, wherein the two strands of the double stranded oligonucleotide are not covalently linked to each other, and wherein each strand of the double stranded oligonucleotide comprises a nucleotide sequence that is complementary to the same nucleotide sequence in a target nucleic acid molecule or a portion thereof (e.g., PTP-1B RNA target). In another embodiment, the two strands of the double stranded oligonucleotide share an identical nucleotide sequence of at least about 15, preferably at least about 16, 17, 18, 19, 20, or 21 nucleotides. 
     In one embodiment, a DFO molecule of the invention comprises a structure having Formula DFO-I: 
                            5′-p-X Z X′-3′            
wherein Z comprises a palindromic or repeat nucleic acid sequence optionally with one or more modified nucleotides (e.g., nucleotide with a modified base, such as 2-amino purine, 2-amino-1,6-dihydro purine or a universal base), for example of length about 2 to about 24 nucleotides in even numbers (e.g., about 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, or 22 or 24 nucleotides), X represents a nucleic acid sequence, for example of length of about 1 to about 21 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 nucleotides), X′ comprises a nucleic acid sequence, for example of length about 1 and about 21 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 nucleotides) having nucleotide sequence complementarity to sequence X or a portion thereof, p comprises a terminal phosphate group that can be present or absent, and wherein sequence X and Z, either independently or together, comprise nucleotide sequence that is complementary to a target nucleic acid sequence or a portion thereof and is of length sufficient to interact (e.g., base pair) with the target nucleic acid sequence or a portion thereof (e.g., PTP-1B RNA target). For example, X independently can comprise a sequence from about 12 to about 21 or more (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more) nucleotides in length that is complementary to nucleotide sequence in a target PTP-1B RNA or a portion thereof. In another non-limiting example, the length of the nucleotide sequence of X and Z together, when X is present, that is complementary to the target RNA or a portion thereof (e.g., PTP-1B RNA target) is from about 12 to about 21 or more nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more). In yet another non-limiting example, when X is absent, the length of the nucleotide sequence of Z that is complementary to the target PTP-1B RNA or a portion thereof is from about 12 to about 24 or more nucleotides (e.g., about 12, 14, 16, 18, 20, 22, 24, or more). In one embodiment X, Z and X′ are independently oligonucleotides, where X and/or Z comprises a nucleotide sequence of length sufficient to interact (e.g., base pair) with a nucleotide sequence in the target RNA or a portion thereof (e.g., PTP-1B RNA target). In one embodiment, the lengths of oligonucleotides X and X′ are identical. In another embodiment, the lengths of oligonucleotides X and X′ are not identical. In another embodiment, the lengths of oligonucleotides X and Z, or Z and X′, or X, Z and X′ are either identical or different.
 
     When a sequence is described in this specification as being of “sufficient” length to interact (i.e., base pair) with another sequence, it is meant that the length is such that the number of bonds (e.g., hydrogen bonds) formed between the two sequences is enough to enable the two sequence to form a duplex under the conditions of interest. Such conditions can be in vitro (e.g., for diagnostic or assay purposes) or in vivo (e.g., for therapeutic purposes). It is a simple and routine matter to determine such lengths. 
     In one embodiment, the invention features a double stranded oligonucleotide construct having Formula DFO-I(a): 
                            5′-p-X Z X′-3′                       3′-X′ Z X-p-5′            
wherein Z comprises a palindromic or repeat nucleic acid sequence or palindromic or repeat-like nucleic acid sequence with one or more modified nucleotides (e.g., nucleotides with a modified base, such as 2-amino purine, 2-amino-1,6-dihydro purine or a universal base), for example of length about 2 to about 24 nucleotides in even numbers (e.g., about 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22 or 24 nucleotides), X represents a nucleic acid sequence, for example of length about 1 to about 21 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 nucleotides), X′ comprises a nucleic acid sequence, for example of length about 1 to about 21 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 nucleotides) having nucleotide sequence complementarity to sequence X or a portion thereof, p comprises a terminal phosphate group that can be present or absent, and wherein each X and Z independently comprises a nucleotide sequence that is complementary to a target nucleic acid sequence or a portion thereof (e.g., PTP-1B RNA target) and is of length sufficient to interact with the target nucleic acid sequence of a portion thereof (e.g., PTP-1B RNA target). For example, sequence X independently can comprise a sequence from about 12 to about 21 or more nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more) in length that is complementary to a nucleotide sequence in a target RNA or a portion thereof (e.g., PTP-1B RNA target). In another non-limiting example, the length of the nucleotide sequence of X and Z together (when X is present) that is complementary to the target PTP-1B RNA or a portion thereof is from about 12 to about 21 or more nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more). In yet another non-limiting example, when X is absent, the length of the nucleotide sequence of Z that is complementary to the target PTP-1B RNA or a portion thereof is from about 12 to about 24 or more nucleotides (e.g., about 12, 14, 16, 18, 20, 22, 24 or more). In one embodiment X, Z and X′ are independently oligonucleotides, where X and/or Z comprises a nucleotide sequence of length sufficient to interact (e.g., base pair) with nucleotide sequence in the target RNA or a portion thereof (e.g., PTP-1B RNA target). In one embodiment, the lengths of oligonucleotides X and X′ are identical. In another embodiment, the lengths of oligonucleotides X and X′ are not identical. In another embodiment, the lengths of oligonucleotides X and Z or Z and X′ or X, Z and X′ are either identical or different. In one embodiment, the double stranded oligonucleotide construct of Formula I(a) includes one or more, specifically 1, 2, 3 or 4, mismatches, to the extent such mismatches do not significantly diminish the ability of the double stranded oligonucleotide to inhibit target gene expression.
 
     In one embodiment, a DFO molecule of the invention comprises structure having Formula DFO-II: 
                            5′-p-X X′-3′            
wherein each X and X′ are independently oligonucleotides of length about 12 nucleotides to about 21 nucleotides, wherein X comprises, for example, a nucleic acid sequence of length about 12 to about 21 nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 nucleotides), X′ comprises a nucleic acid sequence, for example of length about 12 to about 21 nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 nucleotides) having nucleotide sequence complementarity to sequence X or a portion thereof, p comprises a terminal phosphate group that can be present or absent, and wherein X comprises a nucleotide sequence that is complementary to a target nucleic acid sequence (e.g., PTP-1B RNA) or a portion thereof and is of length sufficient to interact (e.g., base pair) with the target nucleic acid sequence of a portion thereof. In one embodiment, the length of oligonucleotides X and X′ are identical. In another embodiment the length of oligonucleotides X and X′ are not identical. In one embodiment, length of the oligonucleotides X and X′ are sufficient to form a relatively stable double stranded oligonucleotide.
 
     In one embodiment, the invention features a double stranded oligonucleotide construct having Formula DFO-II(a): 
                            5′-p-X X′-3′                       3′-X′ X-p-5′            
wherein each X and X′ are independently oligonucleotides of length about 12 nucleotides to about 21 nucleotides, wherein X comprises a nucleic acid sequence, for example of length about 12 to about 21 nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 nucleotides), X′ comprises a nucleic acid sequence, for example of length about 12 to about 21 nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 nucleotides) having nucleotide sequence complementarity to sequence X or a portion thereof, p comprises a terminal phosphate group that can be present or absent, and wherein X comprises nucleotide sequence that is complementary to a target nucleic acid sequence or a portion thereof (e.g., PTP-1B RNA target) and is of length sufficient to interact (e.g., base pair) with the target nucleic acid sequence (e.g., PTP-1B RNA) or a portion thereof. In one embodiment, the lengths of oligonucleotides X and X′ are identical. In another embodiment, the lengths of oligonucleotides X and X′ are not identical. In one embodiment, the lengths of the oligonucleotides X and X′ are sufficient to form a relatively stable double stranded oligonucleotide. In one embodiment, the double stranded oligonucleotide construct of Formula II(a) includes one or more, specifically 1, 2, 3 or 4, mismatches, to the extent such mismatches do not significantly diminish the ability of the double stranded oligonucleotide to inhibit target gene expression.
 
     In one embodiment, the invention features a DFO molecule having Formula DFO-1(b): 
                            5′-p-Z-3′            
where Z comprises a palindromic or repeat nucleic acid sequence optionally including one or more non-standard or modified nucleotides (e.g., nucleotide with a modified base, such as 2-amino purine or a universal base) that can facilitate base-pairing with other nucleotides. Z can be, for example, of length sufficient to interact (e.g., base pair) with nucleotide sequence of a target nucleic acid (e.g., PTP-1B RNA) molecule, preferably of length of at least 12 nucleotides, specifically about 12 to about 24 nucleotides (e.g., about 12, 14, 16, 18, 20, 22 or 24 nucleotides). p represents a terminal phosphate group that can be present or absent.
 
     In one embodiment, a DFO molecule having any of Formula DFO-I, DFO-I(a), DFO-I(b), DFO-II(a) or DFO-II can comprise chemical modifications as described herein without limitation, such as, for example, nucleotides having any of Formulae I-VII, stabilization chemistries as described in Table IV, or any other combination of modified nucleotides and non-nucleotides as described in the various embodiments herein. 
     In one embodiment, the palidrome or repeat sequence or modified nucleotide (e.g., nucleotide with a modified base, such as 2-amino purine or a universal base) in Z of DFO constructs having Formula DFO-I, DFO-I(a) and DFO-I(b), comprises chemically modified nucleotides that are able to interact with a portion of the target nucleic acid sequence (e.g., modified base analogs that can form Watson Crick base pairs or non-Watson Crick base pairs). 
     In one embodiment, a DFO molecule of the invention, for example a DFO having Formula DFO-1 or DFO-11, comprises about 15 to about 40 nucleotides (e.g., about 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, or 40 nucleotides). In one embodiment, a DFO molecule of the invention comprises one or more chemical modifications. In a non-limiting example, the introduction of chemically modified nucleotides and/or non-nucleotides into nucleic acid molecules of the invention provides a powerful tool in overcoming potential limitations of in vivo stability and bioavailability inherent to unmodified RNA molecules that are delivered exogenously. For example, the use of chemically modified nucleic acid molecules can enable a lower dose of a particular nucleic acid molecule for a given therapeutic effect since chemically modified nucleic acid molecules tend to have a longer half-life in serum or in cells or tissues. Furthermore, certain chemical modifications can improve the bioavailability and/or potency of nucleic acid molecules by not only enhancing half-life but also facilitating the targeting of nucleic acid molecules to particular organs, cells or tissues and/or improving cellular uptake of the nucleic acid molecules. Therefore, even if the activity of a chemically modified nucleic acid molecule is reduced in vitro as compared to a native/unmodified nucleic acid molecule, for example when compared to an unmodified RNA molecule, the overall activity of the modified nucleic acid molecule can be greater than the native or unmodified nucleic acid molecule due to improved stability, potency, duration of effect, bioavailability and/or delivery of the molecule. 
     Multifunctional or Multi-Targeted siNA Molecules of the Invention 
     In one embodiment, the invention features siNA molecules comprising multifunctional short interfering nucleic acid (multifunctional siNA) molecules that modulate the expression of one or more genes in a biologic system, such as a cell, tissue, or organism. The multifunctional short interfering nucleic acid (multifunctional siNA) molecules of the invention can target more than one region a PTP-1B target nucleic acid sequence or can target sequences of more than one distinct target nucleic acid molecules. The multifunctional siNA molecules of the invention can be chemically synthesized or expressed from transcription units and/or vectors. The multifunctional siNA molecules of the instant invention provide useful reagents and methods for a variety of human applications, therapeutic, cosmetic, diagnostic, agricultural, veterinary, target validation, genomic discovery, genetic engineering and pharmacogenomic applications. 
     Applicant demonstrates herein that certain oligonucleotides, referred to herein for convenience but not limitation as multifunctional short interfering nucleic acid or multifunctional siNA molecules, are potent mediators of sequence specific regulation of gene expression. The multifunctional siNA molecules of the invention are distinct from other nucleic acid sequences known in the art (e.g., siRNA, miRNA, stRNA, shRNA, antisense oligonucleotides, etc.) in that they represent a class of polynucleotide molecules that are designed such that each strand in the multifunctional siNA construct comprises a nucleotide sequence that is complementary to a distinct nucleic acid sequence in one or more target nucleic acid molecules. A single multifunctional siNA molecule (generally a double-stranded molecule) of the invention can thus target more than one (e.g., 2, 3, 4, 5, or more) differing target nucleic acid target molecules. Nucleic acid molecules of the invention can also target more than one (e.g., 2, 3, 4, 5, or more) region of the same target nucleic acid sequence. As such multifunctional siNA molecules of the invention are useful in down regulating or inhibiting the expression of one or more target nucleic acid molecules (e.g., PTP-1B and apolipoprotein gene targets such as apo AI, apo A-IV, apo B, apo C-III, and apo E). By reducing or inhibiting expression of more than one target nucleic acid molecule with one multifunctional siNA construct, multifunctional siNA molecules of the invention represent a class of potent therapeutic agents that can provide simultaneous inhibition of multiple targets within a disease or pathogen related pathway. Such simultaneous inhibition can provide synergistic therapeutic treatment strategies without the need for separate preclinical and clinical development efforts or complex regulatory approval process. 
     Use of multifunctional siNA molecules that target more then one region of a target nucleic acid molecule (e.g., messenger RNA) is expected to provide potent inhibition of gene expression. For example, a single multifunctional siNA construct of the invention can target both conserved and variable regions of a target nucleic acid molecule, such as PTP-1B, apo AI, apo A-IV, apo B, apo C-III and/or apo E target RNA or DNA, thereby allowing down regulation or inhibition of different splice variants encoded by a single gene, or allowing for targeting of both coding and non-coding regions of a target nucleic acid molecule. 
     Generally, double stranded oligonucleotides are formed by the assembly of two distinct oligonucleotides where the oligonucleotide sequence of one strand is complementary to the oligonucleotide sequence of the second strand; such double stranded oligonucleotides are generally assembled from two separate oligonucleotides (e.g., siRNA). Alternately, a duplex can be formed from a single molecule that folds on itself (e.g., shRNA or short hairpin RNA). These double stranded oligonucleotides are known in the art to mediate RNA interference and all have a common feature wherein only one nucleotide sequence region (guide sequence or the antisense sequence) has complementarity to a target nucleic acid sequence, such as PTP-1B, apo AI, apo A-IV, apo B, apo C-III, and/or apo E targets, and the other strand (sense sequence) comprises nucleotide sequence that is homologous to the target nucleic acid sequence. Generally, the antisense sequence is retained in the active RISC complex and guides the RISC to the target nucleotide sequence by means of complementary base-pairing of the antisense sequence with the target sequence for mediating sequence-specific RNA interference. It is known in the art that in some cell culture systems, certain types of unmodified siRNAs can exhibit “off target” effects. It is hypothesized that this off-target effect involves the participation of the sense sequence instead of the antisense sequence of the siRNA in the RISC complex (see for example Schwarz et al., 2003, Cell, 115, 199-208). In this instance the sense sequence is believed to direct the RISC complex to a sequence (off-target sequence) that is distinct from the intended target sequence, resulting in the inhibition of the off-target sequence. In these double stranded nucleic acid molecules, each strand is complementary to a distinct target nucleic acid sequence. However, the off-targets that are affected by these dsRNAs are not entirely predictable and are non-specific. 
     Distinct from the double stranded nucleic acid molecules known in the art, the applicants have developed a novel, potentially cost effective and simplified method of down regulating or inhibiting the expression of more than one target nucleic acid sequence using a single multifunctional siNA construct. The multifunctional siNA molecules of the invention are designed to be double-stranded or partially double stranded, such that a portion of each strand or region of the multifunctional siNA is complementary to a target nucleic acid sequence of choice. As such, the multifunctional siNA molecules of the invention are not limited to targeting sequences that are complementary to each other, but rather to any two differing target nucleic acid sequences. Multifunctional siNA molecules of the invention are designed such that each strand or region of the multifunctional siNA molecule, that is complementary to a given target nucleic acid sequence, is of suitable length (e.g., from about 16 to about 28 nucleotides in length, preferably from about 18 to about 28 nucleotides in length) for mediating RNA interference against the target nucleic acid sequence. The complementarity between the target nucleic acid sequence and a strand or region of the multifunctional siNA must be sufficient (at least about 8 base pairs) for cleavage of the target nucleic acid sequence by RNA interference. multifunctional siNA of the invention is expected to minimize off-target effects seen with certain siRNA sequences, such as those described in (Schwarz et al., supra). 
     It has been reported that dsRNAs of length between 29 base pairs and 36 base pairs (Tuschl et al., International PCT Publication No. WO 02/44321) do not mediate RNAi. One reason these dsRNAs are inactive may be the lack of turnover or dissociation of the strand that interacts with the target RNA sequence, such that the RISC complex is not able to efficiently interact with multiple copies of the target RNA resulting in a significant decrease in the potency and efficiency of the RNAi process. Applicant has surprisingly found that the multifunctional siNAs of the invention can overcome this hurdle and are capable of enhancing the efficiency and potency of RNAi process. As such, in certain embodiments of the invention, multifunctional siNAs of length of about 29 to about 36 base pairs can be designed such that, a portion of each strand of the multifunctional siNA molecule comprises a nucleotide sequence region that is complementary to a target nucleic acid of length sufficient to mediate RNAi efficiently (e.g., about 15 to about 23 base pairs) and a nucleotide sequence region that is not complementary to the target nucleic acid. By having both complementary and non-complementary portions in each strand of the multifunctional siNA, the multifunctional siNA can mediate RNA interference against a target nucleic acid sequence without being prohibitive to turnover or dissociation (e.g., where the length of each strand is too long to mediate RNAi against the respective target nucleic acid sequence). Furthermore, design of multifunctional siNA molecules of the invention with internal overlapping regions allows the multifunctional siNA molecules to be of favorable (decreased) size for mediating RNA interference and of size that is well suited for use as a therapeutic agent (e.g., wherein each strand is independently from about 18 to about 28 nucleotides in length). Non-limiting examples are illustrated in  FIGS. 16-28 . 
     In one embodiment, a multifunctional siNA molecule of the invention comprises a first region and a second region, where the first region of the multifunctional siNA comprises a nucleotide sequence complementary to a nucleic acid sequence of a first target nucleic acid molecule, and the second region of the multifunctional siNA comprises nucleic acid sequence complementary to a nucleic acid sequence of a second target nucleic acid molecule. In one embodiment, a multifunctional siNA molecule of the invention comprises a first region and a second region, where the first region of the multifunctional siNA comprises nucleotide sequence complementary to a nucleic acid sequence of the first region of a target nucleic acid molecule, and the second region of the multifunctional siNA comprises nucleotide sequence complementary to a nucleic acid sequence of a second region of a the target nucleic acid molecule. In another embodiment, the first region and second region of the multifunctional siNA can comprise separate nucleic acid sequences that share some degree of complementarity (e.g., from about 1 to about 10 complementary nucleotides). In certain embodiments, multifunctional siNA constructs comprising separate nucleic acid seqeunces can be readily linked post-synthetically by methods and reagents known in the art and such linked constructs are within the scope of the invention. Alternately, the first region and second region of the multifunctional siNA can comprise a single nucleic acid sequence having some degree of self complementarity, such as in a hairpin or stem-loop structure. Non-limiting examples of such double stranded and hairpin multifunctional short interfering nucleic acids are illustrated in  FIGS. 16 and 17  respectively. These multifunctional short interfering nucleic acids (multifunctional siNAs) can optionally include certain overlapping nucleotide sequence where such overlapping nucleotide sequence is present in between the first region and the second region of the multifunctional siNA (see for example  FIGS. 18 and 19 ). 
     In one embodiment, the invention features a multifunctional short interfering nucleic acid (multifunctional siNA) molecule, wherein each strand of the multifunctional siNA independently comprises a first region of nucleic acid sequence that is complementary to a distinct target nucleic acid sequence and the second region of nucleotide sequence that is not complementary to the target sequence. The target nucleic acid sequence of each strand is in the same target nucleic acid molecule or different target nucleic acid molecules. 
     In another embodiment, the multifunctional siNA comprises two strands, where: (a) the first strand comprises a region having sequence complementarity to a target nucleic acid sequence (complementary region 1) and a region having no sequence complementarity to the target nucleotide sequence (non-complementary region 1); (b) the second strand of the multifunction siNA comprises a region having sequence complementarity to a target nucleic acid sequence that is distinct from the target nucleotide sequence complementary to the first strand nucleotide sequence (complementary region 2), and a region having no sequence complementarity to the target nucleotide sequence of complementary region 2 (non-complementary region 2); (c) the complementary region 1 of the first strand comprises a nucleotide sequence that is complementary to a nucleotide sequence in the non-complementary region 2 of the second strand and the complementary region 2 of the second strand comprises a nucleotide sequence that is complementary to a nucleotide sequence in the non-complementary region 1 of the first strand. The target nucleic acid sequence of complementary region 1 and complementary region 2 is in the same target nucleic acid molecule or different target nucleic acid molecules. 
     In another embodiment, the multifunctional siNA comprises two strands, where: (a) the first strand comprises a region having sequence complementarity to a target nucleic acid sequence derived from a gene, such as PTP-1B, apo AI, apo A-IV, apo B, apo C-III, and/or apo E, (complementary region 1) and a region having no sequence complementarity to the target nucleotide sequence of complementary region 1 (non-complementary region 1); (b) the second strand of the multifunction siNA comprises a region having sequence complementarity to a target nucleic acid sequence derived from a gene that is distinct from the gene of complementary region 1 (complementary region 2), and a region having no sequence complementarity to the target nucleotide sequence of complementary region 2 (non-complementary region 2); (c) the complementary region 1 of the first strand comprises a nucleotide sequence that is complementary to a nucleotide sequence in the non-complementary region 2 of the second strand and the complementary region 2 of the second strand comprises a nucleotide sequence that is complementary to a nucleotide sequence in the non-complementary region 1 of the first strand. 
     In another embodiment, the multifunctional siNA comprises two strands, where: (a) the first strand comprises a region having sequence complementarity to a target nucleic acid sequence derived from a gene, such as PTP-1B, apo AI, apo A-IV, apo B, apo C-III, and/or apo E, (complementary region 1) and a region having no sequence complementarity to the target nucleotide sequence of complementary region 1 (non-complementary region 1); (b) the second strand of the multifunction siNA comprises a region having sequence complementarity to a target nucleic acid sequence distinct from the target nucleic acid sequence of complementary region 1 (complementary region 2), provided, however, that the target nucleic acid sequence for complementary region 1 and target nucleic acid sequence for complementary region 2 are both derived from the same gene, and a region having no sequence complementarity to the target nucleotide sequence of complementary region 2 (non-complementary region 2); (c) the complementary region 1 of the first strand comprises a nucleotide sequence that is complementary to a nucleotide sequence in the non-complementary region 2 of the second strand and the complementary region 2 of the second strand comprises a nucleotide sequence that is complementary to nucleotide sequence in the non-complementary region 1 of the first strand. 
     In one embodiment, the invention features a multifunctional short interfering nucleic acid (multifunctional siNA) molecule, wherein the multifunctional siNA comprises two complementary nucleic acid sequences in which the first sequence comprises a first region having nucleotide sequence complementary to nucleotide sequence within a target nucleic acid molecule, and in which the second seqeunce comprises a first region having nucleotide sequence complementary to a distinct nucleotide sequence within the same target nucleic acid molecule. Preferably, the first region of the first sequence is also complementary to the nucleotide sequence of the second region of the second sequence, and where the first region of the second sequence is complementary to the nucleotide sequence of the second region of the first sequence. 
     In one embodiment, the invention features a multifunctional short interfering nucleic acid (multifunctional siNA) molecule, wherein the multifunctional siNA comprises two complementary nucleic acid sequences in which the first sequence comprises a first region having a nucleotide sequence complementary to a nucleotide sequence within a first target nucleic acid molecule, and in which the second seqeunce comprises a first region having a nucleotide sequence complementary to a distinct nucleotide sequence within a second target nucleic acid molecule. Preferably, the first region of the first sequence is also complementary to the nucleotide sequence of the second region of the second sequence, and where the first region of the second sequence is complementary to the nucleotide sequence of the second region of the first sequence. 
     In one embodiment, the invention features a multifunctional siNA molecule comprising a first region and a second region, where the first region comprises a nucleic acid sequence having about 18 to about 28 nucleotides complementary to a nucleic acid sequence within a first target nucleic acid molecule, and the second region comprises nucleotide sequence having about 18 to about 28 nucleotides complementary to a distinct nucleic acid sequence within a second target nucleic acid molecule. 
     In one embodiment, the invention features a multifunctional siNA molecule comprising a first region and a second region, where the first region comprises nucleic acid sequence having about 18 to about 28 nucleotides complementary to a nucleic acid sequence within a target nucleic acid molecule, and the second region comprises nucleotide sequence having about 18 to about 28 nucleotides complementary to a distinct nucleic acid sequence within the same target nucleic acid molecule. 
     In one embodiment, the invention features a double stranded multifunctional short interfering nucleic acid (multifunctional siNA) molecule, wherein one strand of the multifunctional siNA comprises a first region having nucleotide sequence complementary to a first target nucleic acid sequence, and the second strand comprises a first region having a nucleotide sequence complementary to a second target nucleic acid sequence. The first and second target nucleic acid sequences can be present in separate target nucleic acid molecules or can be different regions within the same target nucleic acid molecule. As such, multifunctional siNA molecules of the invention can be used to target the expression of different genes, splice variants of the same gene, both mutant and conserved regions of one or more gene transcripts, or both coding and non-coding sequences of the same or differing genes or gene transcripts. 
     In one embodiment, a target nucleic acid molecule of the invention encodes a single protein. In another embodiment, a target nucleic acid molecule encodes more than one protein (e.g., 1, 2, 3, 4, 5 or more proteins). As such, a multifunctional siNA construct of the invention can be used to down regulate or inhibit the expression of several proteins. For example, a multifunctional siNA molecule comprising a region in one strand having nucleotide sequence complementarity to a first target nucleic acid sequence derived from a gene encoding one protein and the second strand comprising a region with nucleotide sequence complementarity to a second target nucleic acid sequence present in target nucleic acid molecules derived from genes encoding two or more proteins (e.g., two or more differing PTP-1B, apo AI, apo A-IV, apo B, apo C-III, and/or apo E target sequences) can be used to down regulate, inhibit, or shut down a particular biologic pathway by targeting, for example, two or more targets involved in a biologic pathway. 
     In one embodiment the invention takes advantage of conserved nucleotide sequences present in different isoforms of cytokines or ligands and receptors for the cytokines or ligands. By designing multifunctional siNAs in a manner where one strand includes a sequence that is complementary to a target nucleic acid sequence conserved among various isoforms of a cytokine and the other strand includes sequence that is complementary to a target nucleic acid sequence conserved among the receptors for the cytokine, it is possible to selectively and effectively modulate or inhibit a biological pathway or multiple genes in a biological pathway using a single multifunctional siNA. 
     In one embodiment, a double stranded multifunctional siNA molecule of the invention comprises a structure having Formula MF-I: 
                            5′-p-X Z X′-3′                       3′-Y′ Z Y-p-5′            
wherein each 5′-p-XZX′-3′ and 5′-p-YZY′-3′ are independently an oligonucleotide of length of about 20 nucleotides to about 300 nucleotides, preferably of about 20 to about 200 nucleotides, about 20 to about 100 nucleotides, about 20 to about 40 nucleotides, about 20 to about 40 nucleotides, about 24 to about 38 nucleotides, or about 26 to about 38 nucleotides; XZ comprises a nucleic acid sequence that is complementary to a first target nucleic acid sequence; YZ is an oligonucleotide comprising nucleic acid sequence that is complementary to a second target nucleic acid sequence; Z comprises nucleotide sequence of length about 1 to about 24 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 nucleotides) that is self complimentary; X comprises nucleotide sequence of length about 1 to about 100 nucleotides, preferably about 1 to about 21 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 nucleotides) that is complementary to nucleotide sequence present in region Y′; Y comprises nucleotide sequence of length about 1 to about 100 nucleotides, preferably about 1- about 21 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 nucleotides) that is complementary to nucleotide sequence present in region X′; each p comprises a terminal phosphate group that is independently present or absent; each XZ and YZ is independently of length sufficient to stably interact (i.e., base pair) with the first and second target nucleic acid sequence, respectively, or a portion thereof. For example, each sequence X and Y can independently comprise sequence from about 12 to about 21 or more nucleotides in length (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more) that is complementary to a target nucleotide sequence in different target nucleic acid molecules, such as target RNAs or a portion thereof In another non-limiting example, the length of the nucleotide sequence of X and Z together that is complementary to the first target nucleic acid sequence or a portion thereof is from about 12 to about 21 or more nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more). In another non-limiting example, the length of the nucleotide sequence of Y and Z together, that is complementary to the second target nucleic acid sequence or a portion thereof is from about 12 to about 21 or more nucleotides (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more). In one embodiment, the first target nucleic acid sequence and the second target nucleic acid sequence are present in the same target nucleic acid molecule (e.g., PTP-1B RNA). In another embodiment, the first target nucleic acid sequence and the second target nucleic acid sequence are present in different target nucleic acid molecules (e.g., PTP-1B, apo AI, apo A-IV, apo B, apo C-III, and/or apo E targets). In one embodiment, Z comprises a palindrome or a repeat sequence. In one embodiment, the lengths of oligonucleotides X and X′ are identical. In another embodiment, the lengths of oligonucleotides X and X′ are not identical. In one embodiment, the lengths of oligonucleotides Y and Y′ are identical. In another embodiment, the lengths of oligonucleotides Y and Y′ are not identical. In one embodiment, the double stranded oligonucleotide construct of Formula I(a) includes one or more, specifically 1, 2, 3 or 4, mismatches, to the extent such mismatches do not significantly diminish the ability of the double stranded oligonucleotide to inhibit target gene expression.
 
     In one embodiment, a multifunctional siNA molecule of the invention comprises a structure having Formula MF-II: 
                            5′-p-X X′-3′                       3′-Y′ Y-p-5′            
wherein each 5′-p-XX′-3′ and 5′-p-YY′-3′ are independently an oligonucleotide of length of about 20 nucleotides to about 300 nucleotides, preferably about 20 to about 200 nucleotides, about 20 to about 100 nucleotides, about 20 to about 40 nucleotides, about 20 to about 40 nucleotides, about 24 to about 38 nucleotides, or about 26 to about 38 nucleotides; X comprises a nucleic acid sequence that is complementary to a first target nucleic acid sequence; Y is an oligonucleotide comprising nucleic acid sequence that is complementary to a second target nucleic acid sequence; X comprises a nucleotide sequence of length about 1 to about 100 nucleotides, preferably about 1 to about 21 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21 nucleotides) that is complementary to nucleotide sequence present in region Y′; Y comprises nucleotide sequence of length about 1 to about 100 nucleotides, preferably about 1 to about 21 nucleotides (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 nucleotides) that is complementary to nucleotide sequence present in region X′; each p comprises a terminal phosphate group that is independently present or absent; each X and Y independently is of length sufficient to stably interact (i.e., base pair) with the first and second target nucleic acid sequence, respectively, or a portion thereof. For example, each sequence X and Y can independently comprise sequence from about 12 to about 21 or more nucleotides in length (e.g., about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more) that is complementary to a target nucleotide sequence in different target nucleic acid molecules, such as PTP-1B, apo AI, apo A-IV, apo B, apo C-III, and/or apo E target sequences or a portion thereof. In one embodiment, the first target nucleic acid sequence and the second target nucleic acid sequence are present in the same target nucleic acid molecule (e.g., PTP-1B RNA or DNA). In another embodiment, the first target nucleic acid sequence and the second target nucleic acid sequence are present in different target nucleic acid molecules, such as PTP-1B, apo AI, apo A-IV, apo B, apo C-III, and/or apo E target sequences or a portion thereof In one embodiment, Z comprises a palindrome or a repeat sequence. In one embodiment, the lengths of oligonucleotides X and X′ are identical. In another embodiment, the lengths of oligonucleotides X and X′ are not identical. In one embodiment, the lengths of oligonucleotides Y and Y′ are identical. In another embodiment, the lengths of oligonucleotides Y and Y′ are not identical. In one embodiment, the double stranded oligonucleotide construct of Formula I(a) includes one or more, specifically 1, 2, 3 or 4, mismatches, to the extent such mismatches do not significantly diminish the ability of the double stranded oligonucleotide to inhibit target gene expression.
 
     In one embodiment, a multifunctional siNA molecule of the invention comprises a structure having Formula MF-III: 
                            X    X′                       Y′-W-Y            
wherein each X, X′, Y, and Y′ is independently an oligonucleotide of length of about 15 nucleotides to about 50 nucleotides, preferably about 18 to about 40 nucleotides, or about 19 to about 23 nucleotides; X comprises nucleotide sequence that is complementary to nucleotide sequence present in region Y′; X′ comprises nucleotide sequence that is complementary to nucleotide sequence present in region Y; each X and X′ is independently of length sufficient to stably interact (i.e., base pair) with a first and a second target nucleic acid sequence, respectively, or a portion thereof; W represents a nucleotide or non-nucleotide linker that connects sequences Y′ and Y; and the multifunctional siNA directs cleavage of the first and second target sequence via RNA interference. In one embodiment, the first target nucleic acid sequence and the second target nucleic acid sequence are present in the same target nucleic acid molecule (e.g., PTP-1B RNA). In another embodiment, the first target nucleic acid sequence and the second target nucleic acid sequence are present in different target nucleic acid molecules such as PTP-1B, apo AI, apo A-IV, apo B, apo C-III, and/or apo E target sequences or a portion thereof. In one embodiment, region W connects the 3′-end of sequence Y′ with the 3′-end of sequence Y. In one embodiment, region W connects the 3′-end of sequence Y′ with the 5′-end of sequence Y. In one embodiment, region W connects the 5′-end of sequence Y′ with the 5′-end of sequence Y. In one embodiment, region W connects the 5′-end of sequence Y′ with the 3′-end of sequence Y. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence X. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence X′. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence Y. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence Y′. In one embodiment, W connects sequences Y and Y′ via a biodegradable linker. In one embodiment, W further comprises a conjugate, label, aptamer, ligand, lipid, or polymer.
 
     In one embodiment, a multifunctional siNA molecule of the invention comprises a structure having Formula MF-IV: 
                            X    X′                       Y′-W-Y            
wherein each X, X′, Y, and Y′ is independently an oligonucleotide of length of about 15 nucleotides to about 50 nucleotides, preferably about 18 to about 40 nucleotides, or about 19 to about 23 nucleotides; X comprises nucleotide sequence that is complementary to nucleotide sequence present in region Y′; X′ comprises nucleotide sequence that is complementary to nucleotide sequence present in region Y; each Y and Y′ is independently of length sufficient to stably interact (i.e., base pair) with a first and a second target nucleic acid sequence, respectively, or a portion thereof; W represents a nucleotide or non-nucleotide linker that connects sequences Y′ and Y; and the multifunctional siNA directs cleavage of the first and second target sequence via RNA interference. In one embodiment, the first target nucleic acid sequence and the second target nucleic acid sequence are present in the same target nucleic acid molecule (e.g., PTP-1B RNA). In another embodiment, the first target nucleic acid sequence and the second target nucleic acid sequence are present in different target nucleic acid molecules, such as PTP-1B, apo AI, apo A-IV, apo B, apo C-III, and/or apo E target sequences or a portion thereof. In one embodiment, region W connects the 3′-end of sequence Y′ with the 3′-end of sequence Y. In one embodiment, region W connects the 3′-end of sequence Y′ with the 5′-end of sequence Y. In one embodiment, region W connects the 5′-end of sequence Y′ with the 5′-end of sequence Y. In one embodiment, region W connects the 5′-end of sequence Y′ with the 3′-end of sequence Y. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence X. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence X′. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence Y. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence Y′. In one embodiment, W connects sequences Y and Y′ via a biodegradable linker. In one embodiment, W further comprises a conjugate, label, aptamer, ligand, lipid, or polymer.
 
     In one embodiment, a multifunctional siNA molecule of the invention comprises a structure having Formula MF-V: 
                            X    X′                       Y′-W-Y            
wherein each X, X′, Y, and Y′ is independently an oligonucleotide of length of about 15 nucleotides to about 50 nucleotides, preferably about 18 to about 40 nucleotides, or about 19 to about 23 nucleotides; X comprises nucleotide sequence that is complementary to nucleotide sequence present in region Y′; X′ comprises nucleotide sequence that is complementary to nucleotide sequence present in region Y; each X, X′, Y, or Y′ is independently of length sufficient to stably interact (i.e., base pair) with a first, second, third, or fourth target nucleic acid sequence, respectively, or a portion thereof; W represents a nucleotide or non-nucleotide linker that connects sequences Y′ and Y; and the multifunctional siNA directs cleavage of the first, second, third, and/or fourth target sequence via RNA interference. In one embodiment, the first, second, third and fourth target nucleic acid sequence are all present in the same target nucleic acid molecule (e.g., PTP-1B RNA). In another embodiment, the first, second, third and fourth target nucleic acid sequence are independently present in different target nucleic acid molecules, such as PTP-1B, apo AI, apo A-IV, apo B, apo C-III, and/or apo E target sequences or a portion thereof In one embodiment, region W connects the 3′-end of sequence Y′ with the 3′-end of sequence Y. In one embodiment, region W connects the 3′-end of sequence Y′ with the 5′-end of sequence Y. In one embodiment, region W connects the 5′-end of sequence Y′ with the 5′-end of sequence Y. In one embodiment, region W connects the 5′-end of sequence Y′ with the 3′-end of sequence Y. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence X. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence X′. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence Y. In one embodiment, a terminal phosphate group is present at the 5′-end of sequence Y′. In one embodiment, W connects sequences Y and Y′ via a biodegradable linker. In one embodiment, W further comprises a conjugate, label, aptamer, ligand, lipid, or polymer.
 
     In one embodiment, regions X and Y of multifunctional siNA molecule of the invention (e.g., having any of Formula MF-I-MF-V), are complementary to different target nucleic acid sequences that are portions of the same target nucleic acid molecule. In one embodiment, such target nucleic acid sequences are at different locations within the coding region of a RNA transcript. In one embodiment, such target nucleic acid sequences comprise coding and non-coding regions of the same RNA transcript. In one embodiment, such target nucleic acid sequences comprise regions of alternately spliced transcripts or precursors of such alternately spliced transcripts. 
     In one embodiment, a multifunctional siNA molecule having any of Formula MF-I-MF-V can comprise chemical modifications as described herein without limitation, such as, for example, nucleotides having any of Formulae I-VII described herein, stabilization chemistries as described in Table IV, or any other combination of modified nucleotides and non-nucleotides as described in the various embodiments herein. 
     In one embodiment, the palidrome or repeat sequence or modified nucleotide (e.g., nucleotide with a modified base, such as 2-amino purine or a universal base) in Z of multifunctional siNA constructs having Formula MF-I or MF-II comprises chemically modified nucleotides that are able to interact with a portion of the target nucleic acid sequence (e.g., modified base analogs that can form Watson Crick base pairs or non-Watson Crick base pairs). 
     In one embodiment, a multifunctional siNA molecule of the invention, for example each strand of a multifunctional siNA having MF-I-MF-V, independently comprises about 15 to about 40 nucleotides (e.g., about 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, or 40 nucleotides). In one embodiment, a multifunctional siNA molecule of the invention comprises one or more chemical modifications. In a non-limiting example, the introduction of chemically modified nucleotides and/or non-nucleotides into nucleic acid molecules of the invention provides a powerful tool in overcoming potential limitations of in vivo stability and bioavailability inherent to unmodified RNA molecules that are delivered exogenously. For example, the use of chemically modified nucleic acid molecules can enable a lower dose of a particular nucleic acid molecule for a given therapeutic effect since chemically modified nucleic acid molecules tend to have a longer half-life in serum or in cells or tissues. Furthermore, certain chemical modifications can improve the bioavailability and/or potency of nucleic acid molecules by not only enhancing half-life but also facilitating the targeting of nucleic acid molecules to particular organs, cells or tissues and/or improving cellular uptake of the nucleic acid molecules. Therefore, even if the activity of a chemically modified nucleic acid molecule is reduced in vitro as compared to a native/unmodified nucleic acid molecule, for example when compared to an unmodified RNA molecule, the overall activity of the modified nucleic acid molecule can be greater than the native or unmodified nucleic acid molecule due to improved stability, potency, duration of effect, bioavailability and/or delivery of the molecule. 
     In another embodiment, the invention features multifunctional siNAs, wherein the multifunctional siNAs are assembled from two separate double-stranded siNAs, with one of the ends of each sense strand is tethered to the end of the sense strand of the other siNA molecule, such that the two antisense siNA strands are annealed to their corresponding sense strand that are tethered to each other at one end (see  FIG. 22 ). The tethers or linkers can be nucleotide-based linkers or non-nucleotide based linkers as generally known in the art and as described herein. 
     In one embodiment, the invention features a multifunctional siNA, wherein the multifunctional siNA is assembled from two separate double-stranded siNAs, with the 5′-end of one sense strand of the siNA is tethered to the 5′-end of the sense strand of the other siNA molecule, such that the 5′-ends of the two antisense siNA strands, annealed to their corresponding sense strand that are tethered to each other at one end, point away (in the opposite direction) from each other (see  FIG. 22(A) ). The tethers or linkers can be nucleotide-based linkers or non-nucleotide based linkers as generally known in the art and as described herein. 
     In one embodiment, the invention features a multifunctional siNA, wherein the multifunctional siNA is assembled from two separate double-stranded siNAs, with the 3′-end of one sense strand of the siNA is tethered to the 3′-end of the sense strand of the other siNA molecule, such that the 5′-ends of the two antisense siNA strands, annealed to their corresponding sense strand that are tethered to each other at one end, face each other (see  FIG. 22(B) ). The tethers or linkers can be nucleotide-based linkers or non-nucleotide based linkers as generally known in the art and as described herein. 
     In one embodiment, the invention features a multifunctional siNA, wherein the multifunctional siNA is assembled from two separate double-stranded siNAs, with the 5′-end of one sense strand of the siNA is tethered to the 3′-end of the sense strand of the other siNA molecule, such that the 5′-end of the one of the antisense siNA strands annealed to their corresponding sense strand that are tethered to each other at one end, faces the 3′-end of the other antisense strand (see  FIG. 22  (C-D)). The tethers or linkers can be nucleotide-based linkers or non-nucleotide based linkers as generally known in the art and as described herein. 
     In one embodiment, the invention features a multifunctional siNA, wherein the multifunctional siNA is assembled from two separate double-stranded siNAs, with the 5′-end of one antisense strand of the siNA is tethered to the 3′-end of the antisense strand of the other siNA molecule, such that the 5′-end of the one of the sense siNA strands annealed to their corresponding antisense sense strand that are tethered to each other at one end, faces the 3′-end of the other sense strand (see  FIG. 22(G-H) ). In one embodiment, the linkage between the 5′-end of the first antisense strand and the 3′-end of the second antisense strand is designed in such a way as to be readily cleavable (e.g., biodegradable linker) such that the 5′end of each antisense strand of the multifunctional siNA has a free 5′-end suitable to mediate RNA interference-based cleavage of the target RNA. The tethers or linkers can be nucleotide-based linkers or non-nucleotide based linkers as generally known in the art and as described herein. 
     In one embodiment, the invention features a multifunctional siNA, wherein the multifunctional siNA is assembled from two separate double-stranded siNAs, with the 5′-end of one antisense strand of the siNA is tethered to the 5′-end of the antisense strand of the other siNA molecule, such that the 3′-end of the one of the sense siNA strands annealed to their corresponding antisense sense strand that are tethered to each other at one end, faces the 3′-end of the other sense strand (see  FIG. 22(E) ). In one embodiment, the linkage between the 5′-end of the first antisense strand and the 5′-end of the second antisense strand is designed in such a way as to be readily cleavable (e.g., biodegradable linker) such that the 5′ end of each antisense strand of the multifunctional siNA has a free 5′-end suitable to mediate RNA interference-based cleavage of the target RNA. The tethers or linkers can be nucleotide-based linkers or non-nucleotide based linkers as generally known in the art and as described herein. 
     In one embodiment, the invention features a multifunctional siNA, wherein the multifunctional siNA is assembled from two separate double-stranded siNAs, with the 3′-end of one antisense strand of the siNA is tethered to the 3′-end of the antisense strand of the other siNA molecule, such that the 5′-end of the one of the sense siNA strands annealed to their corresponding antisense sense strand that are tethered to each other at one end, faces the 3′-end of the other sense strand (see  FIG. 22(F) ). In one embodiment, the linkage between the 5′-end of the first antisense strand and the 5′-end of the second antisense strand is designed in such a way as to be readily cleavable (e.g., biodegradable linker) such that the 5′ end of each antisense strand of the multifunctional siNA has a free 5′-end suitable to mediate RNA interference-based cleavage of the target RNA. The tethers or linkers can be nucleotide-based linkers or non-nucleotide based linkers as generally known in the art and as described herein. 
     In any of the above embodiments, a first target nucleic acid sequence or second target nucleic acid sequence can independently comprise PTP-1B, apo AI, apo A-IV, apo B, apo C-III, and/or apo E RNA, DNA or a portion thereof. In one embodiment, the first target nucleic acid sequence is a PTP-1B RNA, DNA or a portion thereof and the second target nucleic acid sequence is a PTP-1B RNA, DNA of a portion thereof. In one embodiment, the first target nucleic acid sequence is a PTP-1B RNA, DNA or a portion thereof and the second target nucleic acid sequence is a apolipoprotein (e.g., apo AI, apo A-TV, apo B, apo C-III, and/or apo E) RNA, DNA of a portion thereof. 
     Synthesis of Nucleic Acid Molecules 
     Synthesis of nucleic acids greater than 100 nucleotides in length is difficult using automated methods, and the therapeutic cost of such molecules is prohibitive. In this invention, small nucleic acid motifs (“small” refers to nucleic acid motifs no more than 100 nucleotides in length, preferably no more than 80 nucleotides in length, and most preferably no more than 50 nucleotides in length; e.g., individual siNA oligonucleotide sequences or siNA sequences synthesized in tandem) are preferably used for exogenous delivery. The simple structure of these molecules increases the ability of the nucleic acid to invade targeted regions of protein and/or RNA structure. Exemplary molecules of the instant invention are chemically synthesized, and others can similarly be synthesized. 
     Oligonucleotides (e.g., certain modified oligonucleotides or portions of oligonucleotides lacking ribonucleotides) are synthesized using protocols known in the art, for example as described in Caruthers et al., 1992,  Methods in Enzymology  211, 3-19, Thompson et al., International PCT Publication No. WO 99/54459, Wincott et al., 1995,  Nucleic Acids Res.  23, 2677-2684, Wincott et al., 1997,  Methods Mol. Bio.,  74, 59, Brennan et al., 1998,  Biotechnol Bioeng.,  61, 33-45, and Brennan, U.S. Pat. No. 6,001,311. All of these references are incorporated herein by reference. The synthesis of oligonucleotides makes use of common nucleic acid protecting and coupling groups, such as dimethoxytrityl at the 5′-end, and phosphoramidites at the 3′-end. In a non-limiting example, small scale syntheses are conducted on a 394 Applied Biosystems, Inc. synthesizer using a 0.2 μmol scale protocol with a 2.5 min coupling step for 2′-O-methylated nucleotides and a 45 second coupling step for 2′-deoxy nucleotides or 2′-deoxy-2′-fluoro nucleotides. Table V outlines the amounts and the contact times of the reagents used in the synthesis cycle. Alternatively, syntheses at the 0.2 μmol scale can be performed on a 96-well plate synthesizer, such as the instrument produced by Protogene (Palo Alto, Calif.) with minimal modification to the cycle. A 33-fold excess (60 μL of 0.11 M=6.6 μmol) of 2′-O-methyl phosphoramidite and a 105-fold excess of S-ethyl tetrazole (60 μL of 0.25 M=15 μmol) can be used in each coupling cycle of 2′-O-methyl residues relative to polymer-bound 5′-hydroxyl. A 22-fold excess (40 μL of 0.11 M=4.4 μmol) of deoxy phosphoramidite and a 70-fold excess of S-ethyl tetrazole (40 μL of 0.25 M=10 μmol) can be used in each coupling cycle of deoxy residues relative to polymer-bound 5′-hydroxyl. Average coupling yields on the 394 Applied Biosystems, Inc. synthesizer, determined by colorimetric quantitation of the trityl fractions, are typically 97.5-99%. Other oligonucleotide synthesis reagents for the 394 Applied Biosystems, Inc. synthesizer include the following: detritylation solution is 3% TCA in methylene chloride (ABI); capping is performed with 16% N-methyl imidazole in THF (ABI) and 10% acetic anhydride/10% 2,6-lutidine in THF (ABI); and oxidation solution is 16.9 mM I 2 , 49 mM pyridine, 9% water in THF (PerSeptive Biosystems, Inc.). Burdick &amp; Jackson Synthesis Grade acetonitrile is used directly from the reagent bottle. S-Ethyltetrazole solution (0.25 M in acetonitrile) is made up from the solid obtained from American International Chemical, Inc. Alternately, for the introduction of phosphorothioatc linkages, Beaucage reagent (3H-1,2-Benzodithiol-3-one 1,1-dioxide, 0.05 M in acetonitrile) is used. 
     Deprotection of the DNA-based oligonucleotides is performed as follows: the polymer-bound trityl-on oligoribonucleotide is transferred to a 4 mL glass screw top vial and suspended in a solution of 40% aqueous methylamine (1 mL) at 65° C. for 10 minutes. After cooling to −20° C., the supernatant is removed from the polymer support. The support is washed three times with 1.0 mL of EtOH:MeCN:H2O/3:1:1, vortexed and the supernatant is then added to the first supernatant. The combined supernatants, containing the oligoribonucleotide, are dried to a white powder. 
     The method of synthesis used for RNA including certain siNA molecules of the invention follows the procedure as described in Usman et al., 1987,  J. Am. Chem. Soc.,  109, 7845; Scaringe et al., 1990,  Nucleic Acids Res.,  18, 5433; and Wincott et al., 1995,  Nucleic Acids Res.  23, 2677-2684 Wincott et al., 1997,  Methods Mol. Bio.,  74, 59, and makes use of common nucleic acid protecting and coupling groups, such as dimethoxytrityl at the 5′-end, and phosphoramidites at the 3′-end. In a non-limiting example, small scale syntheses are conducted on a 394 Applied Biosystems, Inc. synthesizer using a 0.2 μmol scale protocol with a 7.5 min coupling step for alkylsilyl protected nucleotides and a 2.5 min coupling step for 2′-O-methylated nucleotides. Table V outlines the amounts and the contact times of the reagents used in the synthesis cycle. Alternatively, syntheses at the 0.2 μmol scale can be done on a 96-well plate synthesizer, such as the instrument produced by Protogene (Palo Alto, Calif.) with minimal modification to the cycle. A 33-fold excess (60 ηL of 0.11 M=6.6 μmol) of 2′-O-methyl phosphoramidite and a 75-fold excess of S-ethyl tetrazole (60 μL of 0.25 M=15 μmol) can be used in each coupling cycle of 2′-O-methyl residues relative to polymer-bound 5′-hydroxyl. A 66-fold excess (120 μL of 0.11 M=13.2 μmol) of alkylsilyl (ribo) protected phosphoramidite and a 150-fold excess of S-ethyl tetrazole (120 μL of 0.25 M=30 μmol) can be used in each coupling cycle of ribo residues relative to polymer-bound 5′-hydroxyl. Average coupling yields on the 394 Applied Biosystems, Inc. synthesizer, determined by colorimetric quantitation of the trityl fractions, are typically 97.5-99%. Other oligonucleotide synthesis reagents for the 394 Applied Biosystems, Inc. synthesizer include the following: detritylation solution is 3% TCA in methylene chloride (ABI); capping is performed with 16% N-methyl imidazole in THF (ABI) and 10% acetic anhydride/10% 2,6-lutidine in THF (ABI); oxidation solution is 16.9 mM I 2 , 49 mM pyridine, 9% water in THF (PerSeptive Biosystems, Inc.). Burdick &amp; Jackson Synthesis Grade acetonitrile is used directly from the reagent bottle. S-Ethyltetrazole solution (0.25 M in acetonitrile) is made up from the solid obtained from American International Chemical, Inc. Alternately, for the introduction of phosphorothioate linkages, Beaucage reagent (3H-1,2-Benzodithiol-3-one 1,1-dioxide0.05 M in acetonitrile) is used. 
     Deprotection of the RNA is performed using either a two-pot or one-pot protocol. For the two-pot protocol, the polymer-bound trityl-on oligoribonucleotide is transferred to a 4 mL glass screw top vial and suspended in a solution of 40% aq. methylamine (1 mL) at 65° C. for 10 min. After cooling to −20° C., the supernatant is removed from the polymer support. The support is washed three times with 1.0 mL of EtOH:MeCN:H2O/3:1:1, vortexed and the supernatant is then added to the first supernatant. The combined supernatants, containing the oligoribonucleotide, are dried to a white powder. The base deprotected oligoribonucleotide is resuspended in anhydrous TEA/HF/NMP solution (300 μL of a solution of 1.5 mL N-methylpyrrolidinone, 750 μL TEA and 1 mL TEA·3HF to provide a 1.4 M HF concentration) and heated to 65° C. After 1.5 h, the oligomer is quenched with 1.5 M NH 4 CO 3 . 
     Alternatively, for the one-pot protocol, the polymer-bound trityl-on oligoribonucleotide is transferred to a 4 mL glass screw top vial and suspended in a solution of 33% ethanolic methylamine/DMSO: 1/1 (0.8 mL) at 65° C. for 15 minutes. The vial is brought to room temperature TEA·3HF (0.1 mL) is added and the vial is heated at 65° C. for 15 minutes. The sample is cooled at −20° C. and then quenched with 1.5 M NH 4 HCO 3 . 
     For purification of the trityl-on oligomers, the quenched NH 4 HCO 3  solution is loaded onto a C-18 containing cartridge that had been prewashed with acetonitrile followed by 50 mM TEAA. After washing the loaded cartridge with water, the RNA is detritylated with 0.5% TFA for 13 minutes. The cartridge is then washed again with water, salt exchanged with 1 M NaCl and washed with water again. The oligonucleotide is then eluted with 30% acetonitrile. 
     The average stepwise coupling yields are typically &gt;98% (Wincott et al., 1995  Nucleic Acids Res.  23, 2677-2684). Those of ordinary skill in the art will recognize that the scale of synthesis can be adapted to be larger or smaller than the example described above including but not limited to 96-well format. 
     Alternatively, the nucleic acid molecules of the present invention can be synthesized separately and joined together post-synthetically, for example, by ligation (Moore et al., 1992,  Science  256, 9923; Draper et al., International PCT publication No. WO 93/23569; Shabarova et al., 1991,  Nucleic Acids Research  19, 4247; Bellon et al., 1997,  Nucleosides  &amp;  Nucleotides,  16, 951; Bellon et al., 1997,  Bioconjugate Chem.  8, 204), or by hybridization following synthesis and/or deprotection. 
     The siNA molecules of the invention can also be synthesized via a tandem synthesis methodology as described in Example 1 herein, wherein both siNA strands are synthesized as a single contiguous oligonucleotide fragment or strand separated by a cleavable linker which is subsequently cleaved to provide separate siNA fragments or strands that hybridize and permit purification of the siNA duplex. The linker can be a polynucleotide linker or a non-nucleotide linker. The tandem synthesis of siNA as described herein can be readily adapted to both multiwell/multiplate synthesis platforms such as 96 well or similarly larger multi-well platforms. The tandem synthesis of siNA as described herein can also be readily adapted to large scale synthesis platforms employing batch reactors, synthesis columns and the like. 
     A siNA molecule can also be assembled from two distinct nucleic acid strands or fragments wherein one fragment includes the sense region and the second fragment includes the antisense region of the RNA molecule. 
     The nucleic acid molecules of the present invention can be modified extensively to enhance stability by modification with nuclease resistant groups, for example, 2′-amino, 2′-C-allyl, 2′-fluoro, 2′-O-methyl, 2′-H (for a review see Usman and Cedergren, 1992,  TIBS  17, 34; Usman et al., 1994,  Nucleic Acids Symp. Ser.  31, 163). siNA constructs can be purified by gel electrophoresis using general methods or can be purified by high pressure liquid chromatography (HPLC; see Wincott et al., supra, the totality of which is hereby incorporated herein by reference) and re-suspended in water. 
     In another aspect of the invention, siNA molecules of the invention are expressed from transcription units inserted into DNA or RNA vectors. The recombinant vectors can be DNA plasmids or viral vectors. siNA expressing viral vectors can be constructed based on, but not limited to, adeno-associated virus, retrovirus, adenovirus, or alphavirus. The recombinant vectors capable of expressing the siNA molecules can be delivered as described herein, and persist in target cells. Alternatively, viral vectors can be used that provide for transient expression of siNA molecules. 
     Optimizing Activity of the Nucleic Acid Molecule of the Invention. 
     Chemically synthesizing nucleic acid molecules with modifications (base, sugar and/or phosphate) can prevent their degradation by serum ribonucleases, which can increase their potency (see e.g., Eckstein et al., International Publication No. WO 92/07065; Perrault et al., 1990  Nature  344, 565; Pieken et al., 1991,  Science  253, 314; Usman and Cedergren, 1992,  Trends in Biochem. Sci.  17, 334; Usman et al., International Publication No. WO 93/15187; and Rossi et al., International Publication No. WO 91/03162; Sproat, U.S. Pat. No. 5,334,711; Gold et al., U.S. Pat. No. 6,300,074; and Burgin et al., supra; all of which are incorporated by reference herein). All of the above references describe various chemical modifications that can be made to the base, phosphate and/or sugar moieties of the nucleic acid molecules described herein. Modifications that enhance their efficacy in cells, and removal of bases from nucleic acid molecules to shorten oligonucleotide synthesis times and reduce chemical requirements are desired. 
     There are several examples in the art describing sugar, base and phosphate modifications that can be introduced into nucleic acid molecules with significant enhancement in their nuclease stability and efficacy. For example, oligonucleotides are modified to enhance stability and/or enhance biological activity by modification with nuclease resistant groups, for example, 2′-amino, 2′-C-allyl, 2′-fluoro, 2′-O-methyl, 2′-O-allyl, 2′-H, nucleotide base modifications (for a review see Usman and Cedergren, 1992,  TIBS.  17, 34; Usman et al., 1994,  Nucleic Acids Symp. Ser.  31, 163; Burgin et al., 1996,  Biochemistry,  35, 14090). Sugar modification of nucleic acid molecules have been extensively described in the art (see Eckstein et al., International Publication PCT No. WO 92/07065; Perrault et al.  Nature,  1990, 344, 565-568; Pieken et al.  Science,  1991, 253, 314-317; Usman and Cedergren,  Trends in Biochem. Sci.,  1992, 17, 334-339; Usman et al. International Publication PCT No. WO 93/15187; Sproat, U.S. Pat. No. 5,334,711 and Beigelman et al., 1995,  J. Biol. Chem.,  270, 25702; Beigelman et al., International PCT publication No. WO 97/26270; Beigelman et al., U.S. Pat. No. 5,716,824; Usman et al., U.S. Pat. No. 5,627,053; Woolf et al., International PCT Publication No. WO 98/13526; Thompson et al., U.S. Ser. No. 60/082,404 which was filed on Apr. 20, 1998; Karpeisky et al., 1998,  Tetrahedron Lett.,  39, 1131; Earnshaw and Gait, 1998,  Biopolymers  ( Nucleic Acid Sciences ), 48, 39-55; Verma and Eckstein, 1998,  Annu. Rev. Biochem.,  67, 99-134; and Burlina et al., 1997,  Bioorg. Med. Chem.,  5, 1999-2010; all of the references are hereby incorporated in their totality by reference herein). Such publications describe general methods and strategies to determine the location of incorporation of sugar, base and/or phosphate modifications and the like into nucleic acid molecules without modulating catalysis, and are incorporated by reference herein. In view of such teachings, similar modifications can be used as described herein to modify the siNA nucleic acid molecules of the instant invention so long as the ability of siNA to promote RNAi is cells is not significantly inhibited. 
     While chemical modification of oligonucleotide internucleotide linkages with phosphorothioate, phosphorodithioate, and/or 5′-methylphosphonate linkages improves stability, excessive modifications can cause some toxicity or decreased activity. Therefore, when designing nucleic acid molecules, the amount of these internucleotide linkages should be minimized. The reduction in the concentration of these linkages should lower toxicity, resulting in increased efficacy and higher specificity of these molecules. 
     Short interfering nucleic acid (siNA) molecules having chemical modifications that maintain or enhance activity are provided. Such a nucleic acid is also generally more resistant to nucleases than an unmodified nucleic acid. Accordingly, the in vitro and/or in vivo activity should not be significantly lowered. In cases in which modulation is the goal, therapeutic nucleic acid molecules delivered exogenously should optimally be stable within cells until translation of the target RNA has been modulated long enough to reduce the levels of the undesirable protein. This period of time varies between hours to days depending upon the disease state. Improvements in the chemical synthesis of RNA and DNA (Wincott et al., 1995,  Nucleic Acids Res.  23, 2677; Caruthers et al., 1992,  Methods in Enzymology  211, 3-19 (incorporated by reference herein)) have expanded the ability to modify nucleic acid molecules by introducing nucleotide modifications to enhance their nuclease stability, as described above. 
     In one embodiment, nucleic acid molecules of the invention include one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) G-clamp nucleotides. A G-clamp nucleotide is a modified cytosine analog wherein the modifications confer the ability to hydrogen bond both Watson-Crick and Hoogsteen faces of a complementary guanine within a duplex, see for example Lin and Matteucci, 1998,  J. Am. Chem. Soc.,  120, 8531-8532. A single G-clamp analog substitution within an oligonucleotide can result in substantially enhanced helical thermal stability and mismatch discrimination when hybridized to complementary oligonucleotides. The inclusion of such nucleotides in nucleic acid molecules of the invention results in both enhanced affinity and specificity to nucleic acid targets, complementary sequences, or template strands. In another embodiment, nucleic acid molecules of the invention include one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) LNA “locked nucleic acid” nucleotides such as a 2′,4′-C methylene bicyclo nucleotide (see for example Wengel et al., International PCT Publication No. WO 00/66604 and WO 99/14226). 
     In another embodiment, the invention features conjugates and/or complexes of siNA molecules of the invention. Such conjugates and/or complexes can be used to facilitate delivery of siNA molecules into a biological system, such as a cell. The conjugates and complexes provided by the instant invention can impart therapeutic activity by transferring therapeutic compounds across cellular membranes, altering the pharmacokinetics, and/or modulating the localization of nucleic acid molecules of the invention. The present invention encompasses the design and synthesis of novel conjugates and complexes for the delivery of molecules, including, but not limited to, small molecules, lipids, cholesterol, phospholipids, nucleosides, nucleotides, nucleic acids, antibodies, toxins, negatively charged polymers and other polymers, for example proteins, peptides, hormones, carbohydrates, polyethylene glycols, or polyamines, across cellular membranes. In general, the transporters described are designed to be used either individually or as part of a multi-component system, with or without degradable linkers. These compounds are expected to improve delivery and/or localization of nucleic acid molecules of the invention into a number of cell types originating from different tissues, in the presence or absence of serum (see Sullenger and Cech, U.S. Pat. No. 5,854,038). Conjugates of the molecules described herein can be attached to biologically active molecules via linkers that are biodegradable, such as biodegradable nucleic acid linker molecules. 
     The term “biodegradable linker” as used herein, refers to a nucleic acid or non-nucleic acid linker molecule that is designed as a biodegradable linker to connect one molecule to another molecule, for example, a biologically active molecule to a siNA molecule of the invention or the sense and antisense strands of a siNA molecule of the invention. The biodegradable linker is designed such that its stability can be modulated for a particular purpose, such as delivery to a particular tissue or cell type. The stability of a nucleic acid-based biodegradable linker molecule can be modulated by using various chemistries, for example combinations of ribonucleotides, deoxyribonucleotides, and chemically-modified nucleotides, such as 2′-O-methyl, 2′-fluoro, 2′-amino, 2′-O-amino, 2′-C-allyl, 2′-O-allyl, and other 2′-modified or base modified nucleotides. The biodegradable nucleic acid linker molecule can be a dimer, trimer, tetramer or longer nucleic acid molecule, for example, an oligonucleotide of about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleotides in length, or can comprise a single nucleotide with a phosphorus-based linkage, for example, a phosphoramidate or phosphodiester linkage. The biodegradable nucleic acid linker molecule can also comprise nucleic acid backbone, nucleic acid sugar, or nucleic acid base modifications. 
     The term “biodegradable” as used herein, refers to degradation in a biological system, for example, enzymatic degradation or chemical degradation. 
     The term “biologically active molecule” as used herein refers to compounds or molecules that are capable of eliciting or modifying a biological response in a system. Non-limiting examples of biologically active siNA molecules either alone or in combination with other molecules contemplated by the instant invention include therapeutically active molecules such as antibodies, cholesterol, hormones, antivirals, peptides, proteins, chemotherapeutics, small molecules, vitamins, co-factors, nucleosides, nucleotides, oligonucleotides, enzymatic nucleic acids, antisense nucleic acids, triplex forming oligonucleotides, 2,5-A chimeras, siNA, dsRNA, allozymes, aptamers, decoys and analogs thereof. Biologically active molecules of the invention also include molecules capable of modulating the pharmacokinetics and/or pharmacodynamics of other biologically active molecules, for example, lipids and polymers such as polyamines, polyamides, polyethylene glycol and other polyethers. 
     The term “phospholipid” as used herein, refers to a hydrophobic molecule comprising at least one phosphorus group. For example, a phospholipid can comprise a phosphorus-containing group and saturated or unsaturated alkyl group, optionally substituted with OH, COOH, oxo, amine, or substituted or unsubstituted aryl groups. 
     Therapeutic nucleic acid molecules (e.g., siNA molecules) delivered exogenously optimally are stable within cells until reverse transcription of the RNA has been modulated long enough to reduce the levels of the RNA transcript. The nucleic acid molecules are resistant to nucleases in order to function as effective intracellular therapeutic agents. Improvements in the chemical synthesis of nucleic acid molecules described in the instant invention and in the art have expanded the ability to modify nucleic acid molecules by introducing nucleotide modifications to enhance their nuclease stability as described above. 
     In yet another embodiment, siNA molecules having chemical modifications that maintain or enhance enzymatic activity of proteins involved in RNAi are provided. Such nucleic acids are also generally more resistant to nucleases than unmodified nucleic acids. Thus, in vitro and/or in vivo the activity should not be significantly lowered. 
     Use of the nucleic acid-based molecules of the invention will lead to better treatments by affording the possibility of combination therapies (e.g., multiple siNA molecules targeted to different genes; nucleic acid molecules coupled with known small molecule modulators; or intermittent treatment with combinations of molecules, including different motifs and/or other chemical or biological molecules). The treatment of subjects with siNA molecules can also include combinations of different types of nucleic acid molecules, such as enzymatic nucleic acid molecules (ribozymes), allozymes, antisense, 2,5-A oligoadenylate, decoys, and aptamers. 
     In another aspect a siNA molecule of the invention comprises one or more 5′ and/or a 3′-cap structure, for example, on only the sense siNA strand, the antisense siNA strand, or both siNA strands. 
     By “cap structure” is meant chemical modifications, which have been incorporated at either terminus of the oligonucleotide (see, for example, Adamic et al., U.S. Pat. No. 5,998,203, incorporated by reference herein). These terminal modifications protect the nucleic acid molecule from exonuclease degradation, and may help in delivery and/or localization within a cell. The cap may be present at the 5′-terminus (5′-cap) or at the 3′-terminal (3′-cap) or may be present on both termini. In non-limiting examples, the 5′-cap includes, but is not limited to, glyceryl, inverted deoxy abasic residue (moiety); 4′,5′-methylene nucleotide; 1-(beta-D-erythrofuranosyl) nucleotide, 4′-thio nucleotide; carbocyclic nucleotide; 1,5-anhydrohexitol nucleotide; L-nucleotides; alpha-nucleotides; modified base nucleotide; phosphorodithioate linkage; threo-pentofuranosyl nucleotide; acyclic 3′,4′-seco nucleotide; acyclic 3,4-dihydroxybutyl nucleotide; acyclic 3,5-dihydroxypentyl nucleotide, 3′-3′-inverted nucleotide moiety; 3′-3′-inverted abasic moiety; 3′-2′-inverted nucleotide moiety; 3′-2′-inverted abasic moiety; 1,4-butanediol phosphate; 3′-phosphoramidate; hexylphosphate; aminohexyl phosphate; 3′-phosphate; 3′-phosphorothioate; phosphorodithioate; or bridging or non-bridging methylphosphonate moiety. Non-limiting examples of cap moieties are shown in  FIG. 10 . 
     Non-limiting examples of the 3′-cap include, but are not limited to, glyceryl, inverted deoxy abasic residue (moiety), 4′,5′-methylene nucleotide; 1-(beta-D-erythrofuranosyl) nucleotide; 4′-thio nucleotide, carbocyclic nucleotide; 5′-amino-alkyl phosphate; 1,3-diamino-2-propyl phosphate; 3-aminopropyl phosphate; 6-aminohexyl phosphate; 1,2-aminododecyl phosphate; hydroxypropyl phosphate; 1,5-anhydrohexitol nucleotide; L-nucleotide; alpha-nucleotide; modified base nucleotide; phosphorodithioate; threo-pentofuranosyl nucleotide; acyclic 3′,4′-seco nucleotide; 3,4-dihydroxybutyl nucleotide; 3,5-dihydroxypentyl nucleotide, 5′-5′-inverted nucleotide moiety; 5′-5′-inverted abasic moiety; 5′-phosphoramidate; 5′-phosphorothioate; 1,4-butanediol phosphate; 5′-amino; bridging and/or non-bridging 5′-phosphoramidate, phosphorothioate and/or phosphorodithioate, bridging or non bridging methylphosphonate and 5′-mercapto moieties (for more details see Beaucage and Iyer, 1993,  Tetrahedron  49, 1925; incorporated by reference herein). 
     By the term “non-nucleotide” is meant any group or compound which can be incorporated into a nucleic acid chain in the place of one or more nucleotide units, including either sugar and/or phosphate substitutions, and allows the remaining bases to exhibit their enzymatic activity. The group or compound is abasic in that it does not contain a commonly recognized nucleotide base, such as adenosine, guanine, cytosine, uracil or thymine and therefore lacks a base at the 1′-position. 
     An “alkyl” group refers to a saturated aliphatic hydrocarbon, including straight-chain, branched-chain, and cyclic alkyl groups. Preferably, the alkyl group has 1 to 12 carbons. More preferably, it is a lower alkyl of from 1 to 7 carbons, more preferably 1 to 4 carbons. The alkyl group can be substituted or unsubstituted. When substituted the substituted group(s) is preferably, hydroxyl, cyano, alkoxy, ═O, ═S, NO 2  or N(CH 3 ) 2 , amino, or SH. The term also includes alkenyl groups that are unsaturated hydrocarbon groups containing at least one carbon-carbon double bond, including straight-chain, branched-chain, and cyclic groups. Preferably, the alkenyl group has 1 to 12 carbons. More preferably, it is a lower alkenyl of from 1 to 7 carbons, more preferably 1 to 4 carbons. The alkenyl group may be substituted or unsubstituted. When substituted the substituted group(s) is preferably, hydroxyl, cyano, alkoxy, ═O, ═S, NO 2 , halogen, N(CH 3 ) 2 , amino, or SH. The term “alkyl” also includes alkynyl groups that have an unsaturated hydrocarbon group containing at least one carbon-carbon triple bond, including straight-chain, branched-chain, and cyclic groups. Preferably, the alkynyl group has 1 to 12 carbons. More preferably, it is a lower alkynyl of from 1 to 7 carbons, more preferably 1 to 4 carbons. The alkynyl group may be substituted or unsubstituted. When substituted the substituted group(s) is preferably, hydroxyl, cyano, alkoxy, ═O, ═S, NO 2  or N(CH 3 ) 2 , amino or SH. 
     Such alkyl groups can also include aryl, alkylaryl, carbocyclic aryl, heterocyclic aryl, amide and ester groups. An “aryl” group refers to an aromatic group that has at least one ring having a conjugated pi electron system and includes carbocyclic aryl, heterocyclic aryl and biaryl groups, all of which may be optionally substituted. The preferred substituent(s) of aryl groups are halogen, trihalomethyl, hydroxyl, SH, OH, cyano, alkoxy, alkyl, alkenyl, alkynyl, and amino groups. An “alkylaryl” group refers to an alkyl group (as described above) covalently joined to an aryl group (as described above). Carbocyclic aryl groups are groups wherein the ring atoms on the aromatic ring are all carbon atoms. The carbon atoms are optionally substituted. Heterocyclic aryl groups are groups having from 1 to 3 heteroatoms as ring atoms in the aromatic ring and the remainder of the ring atoms are carbon atoms. Suitable heteroatoms include oxygen, sulfur, and nitrogen, and include furanyl, thienyl, pyridyl, pyrrolyl, N-lower alkyl pyrrolo, pyrimidyl, pyrazinyl, imidazolyl and the like, all optionally substituted. An “amide” refers to an —C(O)—NH—R, where R is either alkyl, aryl, alkylaryl or hydrogen. An “ester” refers to an —C(O)—OR′, where R is either alkyl, aryl, alkylaryl or hydrogen. 
     By “nucleotide” as used herein is as recognized in the art to include natural bases (standard), and modified bases well known in the art. Such bases are generally located at the l′ position of a nucleotide sugar moiety. Nucleotides generally comprise a base, sugar and a phosphate group. The nucleotides can be unmodified or modified at the sugar, phosphate and/or base moiety, (also referred to interchangeably as nucleotide analogs, modified nucleotides, non-natural nucleotides, non-standard nucleotides and other; see, for example, Usman and McSwiggen, supra; Eckstein et al., International PCT Publication No. WO 92/07065; Usman et al., International PCT Publication No. WO 93/15187; Uhlman &amp; Peyman, supra, all are hereby incorporated by reference herein). There are several examples of modified nucleic acid bases known in the art as summarized by Limbach et al., 1994,  Nucleic Acids Res.  22, 2183. Some of the non-limiting examples of base modifications that can be introduced into nucleic acid molecules include, inosine, purine, pyridin-4-one, pyridin-2-one, phenyl, pseudouracil, 2,4,6-trimethoxy benzene, 3-methyl uracil, dihydrouridine, naphthyl, aminophenyl, 5-alkylcytidines (e.g., 5-methylcytidine), 5-alkyluridines (e.g., ribothymidine), 5-halouridine (e.g., 5-bromouridine) or 6-azapyrimidines or 6-alkylpyrimidines (e.g. 6-methyluridine), propyne, and others (Burgin et al., 1996,  Biochemistry,  35, 14090; Uhlman &amp; Peyman, supra). By “modified bases” in this aspect is meant nucleotide bases other than adenine, guanine, cytosine and uracil at 1′ position or their equivalents. 
     In one embodiment, the invention features modified siNA molecules, with phosphate backbone modifications comprising one or more phosphorothioate, phosphorodithioate, methylphosphonate, phosphotriester, morpholino, amidate carbamate, carboxymethyl, acetamidate, polyamide, sulfonate, sulfonamide, sulfamate, formacetal, thioformacetal, and/or alkylsilyl, substitutions. For a review of oligonucleotide backbone modifications, see Hunziker and Leumann, 1995,  Nucleic Acid Analogues: Synthesis and Properties , in  Modern Synthetic Methods , VCH, 331-417, and Mesmaeker et al., 1994,  Novel Backbone Replacements for Oligonucleotides , in  Carbohydrate Modifications in Antisense Research , ACS, 24-39. 
     By “abasic” is meant sugar moieties lacking a base or having other chemical groups in place of a base at the l′ position, see for example Adamic et al., U.S. Pat. No. 5,998,203. 
     By “unmodified nucleoside” is meant one of the bases adenine, cytosine, guanine, thymine, or uracil joined to the 1′ carbon of β-D-ribo-furanose. 
     By “modified nucleoside” is meant any nucleotide base which contains a modification in the chemical structure of an unmodified nucleotide base, sugar and/or phosphate. Non-limiting examples of modified nucleotides are shown by Formulae I-VII and/or other modifications described herein. 
     In connection with 2′-modified nucleotides as described for the present invention, by “amino” is meant 2′-NH 2  or 2′-O—NH 2 , which can be modified or unmodified. Such modified groups are described, for example, in Eckstein et al., U.S. Pat. No. 5,672,695 and Matulic-Adamic et al., U.S. Pat. No. 6,248,878, which are both incorporated by reference in their entireties. 
     Various modifications to nucleic acid siNA structure can be made to enhance the utility of these molecules. Such modifications will enhance shelf-life, half-life in vitro, stability, and ease of introduction of such oligonucleotides to the target site, e.g., to enhance penetration of cellular membranes, and confer the ability to recognize and bind to targeted cells. 
     Administration of Nucleic Acid Molecules 
     A siNA molecule of the invention can be adapted for use to prevent or treat diabetes (e.g., type 1 and/or type 2), obesity, and/or insulin resistance or any other trait, disease or condition that is related to or will respond to the levels of PTP-1B in a cell or tissue, alone or in combination with other therapies. 
     For example, a siNA molecule can comprise a delivery vehicle, including liposomes, for administration to a subject, carriers and diluents and their salts, and/or can be present in pharmaceutically acceptable formulations. Methods for the delivery of nucleic acid molecules are described in Akhtar et al., 1992,  Trends Cell Bio.,  2, 139;  Delivery Strategies for Antisense Oligonucleotide Therapeutics , ed. Akhtar, 1995, Maurer et al., 1999,  Mol. Membr. Biol.,  16, 129-140; Hofland and Huang, 1999,  Handb. Exp. Pharmacol.,  137, 165-192; and Lee et al., 2000,  ACS Symp. Ser.,  752, 184-192, all of which are incorporated herein by reference. Beigelman et al., U.S. Pat. No. 6,395,713 and Sullivan et al., PCT WO 94/02595 further describe the general methods for delivery of nucleic acid molecules. These protocols can be utilized for the delivery of virtually any nucleic acid molecule. Nucleic acid molecules can be administered to cells by a variety of methods known to those of skill in the art, including, but not restricted to, encapsulation in liposomes, by iontophoresis, or by incorporation into other vehicles, such as biodegradable polymers, hydrogels, cyclodextrins (see for example Gonzalez et al., 1999,  Bioconjugate Chem.,  10, 1068-1074; Wang et al., International PCT publication Nos. WO 03/47518 and WO 03/46185), poly(lactic-co-glycolic)acid (PLGA) and PLCA microspheres (see for example U.S. Pat. No. 6,447,796 and US Patent Application Publication No. US 2002130430), biodegradable nanocapsules, and bioadhesive microspheres, or by proteinaceous vectors (O&#39;Hare and Normand, International PCT Publication No. WO 00/53722). In another embodiment, the nucleic acid molecules of the invention can also be formulated or complexed with polyethyleneimine and derivatives thereof, such as polyethyleneimine-polyethyleneglycol-N-acetylgalactosamine (PEI-PEG-GAL) or polyethyleneimine-polyethyleneglycol-tri-N-acetylgalactosamine (PEI-PEG-triGAL) derivatives. In one embodiment, the nucleic acid molecules of the invention are formulated as described in United States Patent Application Publication No. 20030077829, incorporated by reference herein in its entirety. 
     In one embodiment, a siNA molecule of the invention is complexed with membrane disruptive agents such as those described in U.S. Patent Application Publication No. 20010007666, incorporated by reference herein in its entirety including the drawings. In another embodiment, the membrane disruptive agent or agents and the siNA molecule are also complexed with a cationic lipid or helper lipid molecule, such as those lipids described in U.S. Pat. No. 6,235,310, incorporated by reference herein in its entirety including the drawings. 
     In one embodiment, a siNA molecule of the invention is complexed with delivery systems as described in U.S. Patent Application Publication No. 2003077829 and International PCT Publication Nos. WO 00/03683 and WO 02/087541, all incorporated by reference herein in their entirety including the drawings. 
     In one embodiment, the nucleic acid molecules of the invention are administered via pulmonary delivery, such as by inhalation of an aerosol or spray dried formulation administered by an inhalation device or nebulizer, providing rapid local uptake of the nucleic acid molecules into relevant pulmonary tissues. Solid particulate compositions containing respirable dry particles of micronized nucleic acid compositions can be prepared by grinding dried or lyophilized nucleic acid compositions, and then passing the micronized composition through, for example, a 400 mesh screen to break up or separate out large agglomerates. A solid particulate composition comprising the nucleic acid compositions of the invention can optionally contain a dispersant which serves to facilitate the formation of an aerosol as well as other therapeutic compounds. A suitable dispersant is lactose, which can be blended with the nucleic acid compound in any suitable ratio, such as a 1 to 1 ratio by weight. 
     Aerosols of liquid particles comprising a nucleic acid composition of the invention can be produced by any suitable means, such as with a nebulizer (see for example U.S. Pat. No. 4,501,729). Nebulizers are commercially available devices which transform solutions or suspensions of an active ingredient into a therapeutic aerosol mist either by means of acceleration of a compressed gas, typically air or oxygen, through a narrow venturi orifice or by means of ultrasonic agitation. Suitable formulations for use in nebulizers comprise the active ingredient in a liquid carrier in an amount of up to 40% w/w preferably less than 20% w/w of the formulation. The carrier is typically water or a dilute aqueous alcoholic solution, preferably made isotonic with body fluids by the addition of, for example, sodium chloride or other suitable salts. Optional additives include preservatives if the formulation is not prepared sterile, for example, methyl hydroxybenzoate, anti-oxidants, flavorings, volatile oils, buffering agents and emulsifiers and other formulation surfactants. The aerosols of solid particles comprising the active composition and surfactant can likewise be produced with any solid particulate aerosol generator. Aerosol generators for administering solid particulate therapeutics to a subject produce particles which are respirable, as explained above, and generate a volume of aerosol containing a predetermined metered dose of a therapeutic composition at a rate suitable for human administration. 
     In one embodiment, a solid particulate aerosol generator of the invention is an insufflator. Suitable formulations for administration by insufflation include finely comminuted powders which can be delivered by means of an insufflator. In the insufflator, the powder, e.g., a metered dose thereof effective to carry out the treatments described herein, is contained in capsules or cartridges, typically made of gelatin or plastic, which are either pierced or opened in situ and the powder delivered by air drawn through the device upon inhalation or by means of a manually-operated pump. The powder employed in the insufflator consists either solely of the active ingredient or of a powder blend comprising the active ingredient, a suitable powder diluent, such as lactose, and an optional surfactant. The active ingredient typically comprises from 0.1 to 100 w/w of the formulation. A second type of illustrative aerosol generator comprises a metered dose inhaler. Metered dose inhalers are pressurized aerosol dispensers, typically containing a suspension or solution formulation of the active ingredient in a liquified propellant. During use these devices discharge the formulation through a valve adapted to deliver a metered volume to produce a fine particle spray containing the active ingredient. Suitable propellants include certain chlorofluorocarbon compounds, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane and mixtures thereof. The formulation can additionally contain one or more co-solvents, for example, ethanol, emulsifiers and other formulation surfactants, such as oleic acid or sorbitan trioleate, anti-oxidants and suitable flavoring agents. Other methods for pulmonary delivery are described in, for example US Patent Application No. 20040037780, and U.S. Pat. Nos. 6,592,904; 6,582,728; 6,565,885, all incorporated by reference herein. 
     In one embodiment, the invention features the use of methods to deliver the nucleic acid molecules of the instant invention to the central nervous system and/or peripheral nervous system. Experiments have demonstrated the efficient in vivo uptake of nucleic acids by neurons. As an example of local administration of nucleic acids to nerve cells, Sommer et al., 1998,  Antisense Nuc. Acid Drug Dev.,  8, 75, describe a study in which a 15 mer phosphorothioate antisense nucleic acid molecule to c-fos is administered to rats via microinjection into the brain. Antisense molecules labeled with tetramethylrhodamine-isothiocyanate (TRITC) or fluorescein isothiocyanate (FITC) were taken up by exclusively by neurons thirty minutes post-injection. A diffuse cytoplasmic staining and nuclear staining was observed in these cells. As an example of systemic administration of nucleic acid to nerve cells, Epa et al., 2000,  Antisense Nuc. Acid Drug Dev.,  10, 469, describe an in vivo mouse study in which beta-cyclodextrin-adamantane-oligonucleotide conjugates were used to target the p75 neurotrophin receptor in neuronally differentiated PC12 cells. Following a two week course of IP administration, pronounced uptake of p75 neurotrophin receptor antisense was observed in dorsal root ganglion (DRG) cells. In addition, a marked and consistent down-regulation of p′75 was observed in DRG neurons. Additional approaches to the targeting of nucleic acid to neurons are described in Broaddus et al., 1998,  J. Neurosurg.,  88(4), 734; Karle et al., 1997,  Eur. J. Pharmocol.,  340(2/3), 153; Bannai et al., 1998,  Brain Research,  784(1,2), 304; Rajakumar et al., 1997,  Synapse,  26(3), 199; Wu-pong et al., 1999,  BioPharm,  12(1), 32; Bannai et al., 1998,  Brain Res. Protoc.,  3(1), 83; Simantov et al., 1996,  Neuroscience,  74(1), 39. Nucleic acid molecules of the invention are therefore amenable to delivery to and uptake by cells that express repeat expansion allelic variants for modulation of RE gene expression. The delivery of nucleic acid molecules of the invention, targeting RE is provided by a variety of different strategies. Traditional approaches to CNS delivery that can be used include, but are not limited to, intrathecal and intracerebroventricular administration, implantation of catheters and pumps, direct injection or perfusion at the site of injury or lesion, injection into the brain arterial system, or by chemical or osmotic opening of the blood-brain barrier. Other approaches can include the use of various transport and carrier systems, for example though the use of conjugates and biodegradable polymers. Furthermore, gene therapy approaches, for example as described in Kaplitt et al., U.S. Pat. No. 6,180,613 and Davidson, WO 04/013280, can be used to express nucleic acid molecules in the CNS. 
     In one embodiment, nucleic acid molecules of the invention are administered to the central nervous system (CNS) or peripheral nervous system (PNS). Experiments have demonstrated the efficient in vivo uptake of nucleic acids by neurons. As an example of local administration of nucleic acids to nerve cells, Sommer et al., 1998,  Antisense Nuc. Acid Drug Dev.,  8, 75, describe a study in which a 15 mer phosphorothioate antisense nucleic acid molecule to c-fos is administered to rats via microinjection into the brain. Antisense molecules labeled with tetramethylrhodamine-isothiocyanate (TRITC) or fluorescein isothiocyanate (FITC) were taken up by exclusively by neurons thirty minutes post-injection. A diffuse cytoplasmic staining and nuclear staining was observed in these cells. As an example of systemic administration of nucleic acid to nerve cells, Epa et al., 2000,  Antisense Nuc. Acid Drug Dev.,  10, 469, describe an in vivo mouse study in which beta-cyclodextrin-adamantane-oligonucleotide conjugates were used to target the p75 neurotrophin receptor in neuronally differentiated PC12 cells. Following a two week course of IP administration, pronounced uptake of p75 neurotrophin receptor antisense was observed in dorsal root ganglion (DRG) cells. In addition, a marked and consistent down-regulation of p75 was observed in DRG neurons. Additional approaches to the targeting of nucleic acid to neurons are described in Broaddus et al., 1998,  J. Neurosurg.,  88(4), 734; Karle et al., 1997,  Eur. J. Pharmocol.,  340(2/3), 153; Bannai et al., 1998,  Brain Research,  784(1,2), 304; Rajakumar et al., 1997,  Synapse,  26(3), 199; Wu-pong et al., 1999,  BioPharm,  12(1), 32; Bannai et al., 1998,  Brain Res. Protoc.,  3(1), 83; Simantov et al., 1996,  Neuroscience,  74(1), 39. Nucleic acid molecules of the invention are therefore amenable to delivery to and uptake by cells in the CNS and/or PNS. 
     The delivery of nucleic acid molecules of the invention to the CNS is provided by a variety of different strategies. Traditional approaches to CNS delivery that can be used include, but are not limited to, intrathecal and intracerebroventricular administration, implantation of catheters and pumps, direct injection or perfusion at the site of injury or lesion, injection into the brain arterial system, or by chemical or osmotic opening of the blood-brain barrier. Other approaches can include the use of various transport and carrier systems, for example though the use of conjugates and biodegradable polymers. Furthermore, gene therapy approaches, for example as described in Kaplitt et al., U.S. Pat. No. 6,180,613 and Davidson, WO 04/013280, can be used to express nucleic acid molecules in the CNS. 
     In one embodiment, delivery systems of the invention include, for example, aqueous and nonaqueous gels, creams, multiple emulsions, microcmulsions, liposomes, ointments, aqueous and nonaqueous solutions, lotions, aerosols, hydrocarbon bases and powders, and can contain excipients such as solubilizers, permeation enhancers (e.g., fatty acids, fatty acid esters, fatty alcohols and amino acids), and hydrophilic polymers (e.g., polycarbophil and polyvinylpyrolidone). In one embodiment, the pharmaceutically acceptable carrier is a liposome or a transdermal enhancer. Examples of liposomes which can be used in this invention include the following: (1) CellFectin, 1:1.5 (M/M) liposome formulation of the cationic lipid N,NI,NII,NIII-tetramethyl-N,NI,NII,NIII-tetrapalmit-y-spermine and dioleoyl phosphatidylethanolamine (DOPE) (GIBCO BRL); (2) Cytofectin GS V, 2:1 (M/M) liposome formulation of a cationic lipid and DOPE (Glen Research); (3) DOTAP (N-[1-(2,3-dioleoyloxy)-N,N,N-tri-methyl-ammoniummethylsulfate) (Boehringer Manheim); and (4) Lipofectamine, 3:1 (M/M) liposome formulation of the polycationic lipid DOSPA and the neutral lipid DOPE (GIBCO BRL). 
     In one embodiment, delivery systems of the invention include patches, tablets, suppositories, pessaries, gels and creams, and can contain excipients such as solubilizers and enhancers (e.g., propylene glycol, bile salts and amino acids), and other vehicles (e.g., polyethylene glycol, fatty acid esters and derivatives, and hydrophilic polymers such as hydroxypropylmethylcellulose and hyaluronic acid). 
     In one embodiment, a siNA molecule of the invention is administered iontophoretically, for example to the dermis. Non-limiting examples of iontophoretic delivery are described in, for example, WO 03/043689 and WO 03/030989, which are incorporated by reference in their entireties herein. 
     In one embodiment, siNA molecules of the invention are formulated or complexed with polyethylenimine (e.g., linear or branched PEI) and/or polyethylenimine derivatives, including for example grafted PETS such as galactose PET, cholesterol PET, antibody derivatized PEI, and polyethylene glycol PEI (PEG-PEI) derivatives thereof (see for example Ogris et al., 2001,  AAPA PharmSci,  3, 1-11; Furgeson et al., 2003, Bioconjugate Chem., 14, 840-847; Kunath et al., 2002, Phramaceutical Research, 19, 810-817; Choi et al., 2001, Bull. Korean Chem. Soc., 22, 46-52; Bettinger et al., 1999, Bioconjugate Chem., 10, 558-561; Peterson et al., 2002, Bioconjugate Chem., 13, 845-854; Erbacher et al., 1999, Journal of Gene Medicine Preprint, 1, 1-18; Godbey et al., 1999., PNAS USA, 96, 5177-5181; Godbey et al., 1999, Journal of Controlled Release, 60, 149-160; Diebold et al., 1999, Journal of Biological Chemistry, 274, 19087-19094; Thomas and Klibanov, 2002, PNAS USA, 99, 14640-14645; and Sagara, U.S. Pat. No. 6,586,524, incorporated by reference herein. 
     In one embodiment, a siNA molecule of the invention comprises a bioconjugate, for example a nucleic acid conjugate as described in Vargeese et al., U.S. Ser. No. 10/427,160, filed Apr. 30, 2003; U.S. Pat. No. 6,528,631; U.S. Pat. No. 6,335,434; U.S. Pat. No. 6,235,886; U.S. Pat. No. 6,153,737; U.S. Pat. No. 5,214,136; U.S. Pat. No. 5,138,045, all incorporated by reference herein. 
     Thus, the invention features a pharmaceutical composition comprising one or more nucleic acid(s) of the invention in an acceptable carrier, such as a stabilizer, buffer, and the like. The polynucleotides of the invention can be administered (e.g., RNA, DNA or protein) and introduced to a subject by any standard means, with or without stabilizers, buffers, and the like, to form a pharmaceutical composition. When it is desired to use a liposome delivery mechanism, standard protocols for formation of liposomes can be followed. The compositions of the present invention can also be formulated and used as creams, gels, sprays, oils and other suitable compositions for topical, dermal, or transdermal administration as is known in the art. 
     The present invention also includes pharmaceutically acceptable formulations of the compounds described. These formulations include salts of the above compounds, e.g., acid addition salts, for example, salts of hydrochloric, hydrobromic, acetic acid, and benzene sulfonic acid. 
     A pharmacological composition or formulation refers to a composition or formulation in a form suitable for administration, e.g., systemic or local administration, into a cell or subject, including for example a human. Suitable forms, in part, depend upon the use or the route of entry, for example oral, transdermal, or by injection. Such forms should not prevent the composition or formulation from reaching a target cell (i.e., a cell to which the negatively charged nucleic acid is desirable for delivery). For example, pharmacological compositions injected into the blood stream should be soluble. Other factors are known in the art, and include considerations such as toxicity and forms that prevent the composition or formulation from exerting its effect. 
     In one embodiment, siNA molecules of the invention are administered to a subject by systemic administration in a pharmaceutically acceptable composition or formulation. By “systemic administration” is meant in vivo systemic absorption or accumulation of drugs in the blood stream followed by distribution throughout the entire body. Administration routes that lead to systemic absorption include, without limitation: intravenous, subcutaneous, portal vein, intraperitoneal, inhalation, oral, intrapulmonary and intramuscular. Each of these administration routes exposes the siNA molecules of the invention to an accessible diseased tissue. The rate of entry of a drug into the circulation has been shown to be a function of molecular weight or size. The use of a liposome or other drug carrier comprising the compounds of the instant invention can potentially localize the drug, for example, in certain tissue types, such as the tissues of the reticular endothelial system (RES). A liposome formulation that can facilitate the association of drug with the surface of cells, such as, lymphocytes and macrophages is also useful. This approach can provide enhanced delivery of the drug to target cells by taking advantage of the specificity of macrophage and lymphocyte immune recognition of abnormal cells. 
     By “pharmaceutically acceptable formulation” or “pharmaceutically acceptable composition” is meant, a composition or formulation that allows for the effective distribution of the nucleic acid molecules of the instant invention in the physical location most suitable for their desired activity. Non-limiting examples of agents suitable for formulation with the nucleic acid molecules of the instant invention include: P-glycoprotein inhibitors (such as Pluronic P85); biodegradable polymers, such as poly (DL-lactide-coglycolide) microspheres for sustained release delivery (Emerich, D F et al, 1999,  Cell Transplant,  8, 47-58); and loaded nanoparticles, such as those made of polybutylcyanoacrylate. Other non-limiting examples of delivery strategies for the nucleic acid molecules of the instant invention include material described in Boado et al., 1998,  J. Pharm. Sci.,  87, 1308-1315; Tyler et al., 1999,  FEBS Lett.,  421, 280-284; Pardridge et al., 1995,  PNAS USA.,  92, 5592-5596; Boado, 1995,  Adv. Drug Delivery Rev.,  15, 73-107; Aldrian-Herrada et al., 1998,  Nucleic Acids Res.,  26, 4910-4916; and Tyler et al., 1999,  PNAS USA.,  96, 7053-7058. 
     The invention also features the use of a composition comprising surface-modified liposomes containing poly (ethylene glycol) lipids (PEG-modified, or long-circulating liposomes or stealth liposomes) and nucleic acid molecules of the invention. These formulations offer a method for increasing the accumulation of drugs (e.g., siNA) in target tissues. This class of drug carriers resists opsonization and elimination by the mononuclear phagocytic system (MPS or RES), thereby enabling longer blood circulation times and enhanced tissue exposure for the encapsulated drug (Lasic et al.  Chem. Rev.  1995, 95, 2601-2627; Ishiwata et al.,  Chem. Pharm. Bull.  1995, 43, 1005-1011). Such liposomes have been shown to accumulate selectively in tumors, presumably by extravasation and capture in the neovascularized target tissues (Lasic et al.,  Science  1995, 267, 1275-1276; Oku et al., 1995,  Biochim. Biophys. Acta,  1238, 86-90). The long-circulating liposomes enhance the pharmacokinetics and pharmacodynamics of DNA and RNA, particularly compared to conventional cationic liposomes which are known to accumulate in tissues of the MPS (Liu et al.,  J. Biol. Chem.  1995, 42, 24864-24870; Choi et al., International PCT Publication No. WO 96/10391; Ansell et al., International PCT Publication No. WO 96/10390; Holland et al., International PCT Publication No. WO 96/10392). Long-circulating liposomes are also likely to protect drugs from nuclease degradation to a greater extent compared to cationic liposomes, based on their ability to avoid accumulation in metabolically aggressive MPS tissues such as the liver and spleen. 
     The present invention also includes compositions prepared for storage or administration that include a pharmaceutically effective amount of the desired compounds in a pharmaceutically acceptable carrier or diluent. Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical art, and are described, for example, in  Remington&#39;s Pharmaceutical Sciences , Mack Publishing Co. (A. R. Gennaro edit. 1985), hereby incorporated by reference herein. For example, preservatives, stabilizers, dyes and flavoring agents can be provided. These include sodium benzoate, sorbic acid and esters of p-hydroxybenzoic acid. In addition, antioxidants and suspending agents can be used. 
     A pharmaceutically effective dose is that dose required to prevent, inhibit the occurrence, or treat (alleviate a symptom to some extent, preferably all of the symptoms) of a disease state. The pharmaceutically effective dose depends on the type of disease, the composition used, the route of administration, the type of mammal being treated, the physical characteristics of the specific mammal under consideration, concurrent medication, and other factors that those skilled in the medical arts will recognize. Generally, an amount between 0.1 mg/kg and 100 mg/kg body weight/day of active ingredients is administered dependent upon potency of the negatively charged polymer. 
     The nucleic acid molecules of the invention and formulations thereof can be administered orally, topically, parenterally, by inhalation or spray, or rectally in dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants and/or vehicles. The term parenteral as used herein includes percutaneous, subcutaneous, intravascular (e.g., intravenous), intramuscular, or intrathecal injection or infusion techniques and the like. In addition, there is provided a pharmaceutical formulation comprising a nucleic acid molecule of the invention and a pharmaceutically acceptable carrier. One or more nucleic acid molecules of the invention can be present in association with one or more non-toxic pharmaceutically acceptable carriers and/or diluents and/or adjuvants, and if desired other active ingredients. The pharmaceutical compositions containing nucleic acid molecules of the invention can be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion, hard or soft capsules, or syrups or elixirs. 
     Compositions intended for oral use can be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions can contain one or more such sweetening agents, flavoring agents, coloring agents or preservative agents in order to provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients that are suitable for the manufacture of tablets. These excipients can be, for example, inert diluents; such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, corn starch, or alginic acid; binding agents, for example starch, gelatin or acacia; and lubricating agents, for example magnesium stearate, stearic acid or talc. The tablets can be uncoated or they can be coated by known techniques. In some cases such coatings can be prepared by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monosterate or glyceryl distearate can be employed. 
     Formulations for oral use can also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example peanut oil, liquid paraffin or olive oil. 
     Aqueous suspensions contain the active materials in a mixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydropropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents can be a naturally-occurring phosphatide, for example, lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions can also contain one or more preservatives, for example ethyl, or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin. 
     Oily suspensions can be formulated by suspending the active ingredients in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions can contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents and flavoring agents can be added to provide palatable oral preparations. These compositions can be preserved by the addition of an anti-oxidant such as ascorbic acid 
     Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents or suspending agents are exemplified by those already mentioned above. Additional excipients, for example sweetening, flavoring and coloring agents, can also be present. 
     Pharmaceutical compositions of the invention can also be in the form of oil-in-water emulsions. The oily phase can be a vegetable oil or a mineral oil or mixtures of these. Suitable emulsifying agents can be naturally-occurring gums, for example gum acacia or gum tragacanth, naturally-occurring phosphatides, for example soy bean, lecithin, and esters or partial esters derived from fatty acids and hexitol, anhydrides, for example sorbitan monooleate, and condensation products of the said partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate. The emulsions can also contain sweetening and flavoring agents. 
     Syrups and elixirs can be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol, glucose or sucrose. Such formulations can also contain a demulcent, a preservative and flavoring and coloring agents. The pharmaceutical compositions can be in the form of a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents that have been mentioned above. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parentally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer&#39;s solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables. 
     The nucleic acid molecules of the invention can also be administered in the form of suppositories, e.g., for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials include cocoa butter and polyethylene glycols. 
     Nucleic acid molecules of the invention can be administered parenterally in a sterile medium. The drug, depending on the vehicle and concentration used, can either be suspended or dissolved in the vehicle. Advantageously, adjuvants such as local anesthetics, preservatives and buffering agents can be dissolved in the vehicle. 
     Dosage levels of the order of from about 0.1 mg to about 140 mg per kilogram of body weight per day are useful in the treatment of the above-indicated conditions (about 0.5 mg to about 7 g per subject per day). The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form varies depending upon the host treated and the particular mode of administration. Dosage unit forms generally contain between from about 1 mg to about 500 mg of an active ingredient. 
     It is understood that the specific dose level for any particular subject depends upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, and rate of excretion, drug combination and the severity of the particular disease undergoing therapy. 
     For administration to non-human animals, the composition can also be added to the animal feed or drinking water. It can be convenient to formulate the animal feed and drinking water compositions so that the animal takes in a therapeutically appropriate quantity of the composition along with its diet. It can also be convenient to present the composition as a premix for addition to the feed or drinking water. 
     The nucleic acid molecules of the present invention can also be administered to a subject in combination with other therapeutic compounds to increase the overall therapeutic effect. The use of multiple compounds to treat an indication can increase the beneficial effects while reducing the presence of side effects. 
     In one embodiment, the invention comprises compositions suitable for administering nucleic acid molecules of the invention to specific cell types. For example, the asialoglycoprotein receptor (ASGPr) (Wu and Wu, 1987,  J. Biol. Chem.  262, 4429-4432) is unique to hepatocytes and binds branched galactose-terminal glycoproteins, such as asialoorosomucoid (ASOR). In another example, the folate receptor is overexpressed in many cancer cells. Binding of such glycoproteins, synthetic glycoconjugates, or folates to the receptor takes place with an affinity that strongly depends on the degree of branching of the oligosaccharide chain, for example, triatennary structures are bound with greater affinity than biatenarry or monoatennary chains (Baenziger and Fiete, 1980,  Cell,  22, 611-620; Connolly et al., 1982,  J. Biol. Chem.,  257, 939-945). Lee and Lee, 1987,  Glycoconjugate J.,  4, 317-328, obtained this high specificity through the use of N-acetyl-D-galactosamine as the carbohydrate moiety, which has higher affinity for the receptor, compared to galactose. This “clustering effect” has also been described for the binding and uptake of mannosyl-terminating glycoproteins or glycoconjugates (Ponpipom et al., 1981,  J. Med. Chem.,  24, 1388-1395). The use of galactose, galactosamine, or folate based conjugates to transport exogenous compounds across cell membranes can provide a targeted delivery approach to, for example, the treatment of liver disease, cancers of the liver, or other cancers. The use of bioconjugates can also provide a reduction in the required dose of therapeutic compounds required for treatment. Furthermore, therapeutic bioavailability, pharmacodynamics, and pharmacokinetic parameters can be modulated through the use of nucleic acid bioconjugates of the invention. Non-limiting examples of such bioconjugates are described in Vargeese et al., U.S. Ser. No. 10/201,394, filed Aug. 13, 2001; and Matulic-Adamic et al., U.S. Ser. No. 60/362,016, filed Mar. 6, 2002. 
     Alternatively, certain siNA molecules of the instant invention can be expressed within cells from eukaryotic promoters (e.g., Izant and Weintraub, 1985,  Science,  229, 345; McGarry and Lindquist, 1986,  Proc. Natl. Acad. Sci., USA  83, 399; Scanlon et al., 1991,  Proc. Math Acad. Sci. USA,  88, 10591-5; Kashani-Sabet et al., 1992,  Antisense Res. Dev.,  2, 3-15; Dropulic et al., 1992,  J. Vivol.,  66, 1432-41; Weerasinghe et al., 1991,  J. Vivol.,  65, 5531-4; Ojwang et al., 1992,  Proc. Natl. Acad. Sci. USA,  89, 10802-6; Chen et al., 1992,  Nucleic Acids Res.,  20, 4581-9; Sarver et al., 1990  Science,  247, 1222-1225; Thompson et al., 1995,  Nucleic Acids Res.,  23, 2259; Good et al., 1997,  Gene Therapy,  4, 45. Those skilled in the art realize that any nucleic acid can be expressed in eukaryotic cells from the appropriate DNA/RNA vector. The activity of such nucleic acids can be augmented by their release from the primary transcript by a enzymatic nucleic acid (Draper et al., PCT WO 93/23569, and Sullivan et al., PCT WO 94/02595; Ohkawa et al., 1992,  Nucleic Acids Symp. Ser.,  27, 15-6; Taira et al., 1991,  Nucleic Acids Res.,  19, 5125-30; Ventura et al., 1993,  Nucleic Acids Res.,  21, 3249-55; Chowrira et al., 1994,  J. Biol. Chem.,  269, 25856. 
     In another aspect of the invention, RNA molecules of the present invention can be expressed from transcription units (see for example Couture et al., 1996,  TIG.,  12, 510) inserted into DNA or RNA vectors. The recombinant vectors can be DNA plasmids or viral vectors. siNA expressing viral vectors can be constructed based on, but not limited to, adeno-associated virus, retrovirus, adenovirus, or alphavirus. In another embodiment, pol III based constructs are used to express nucleic acid molecules of the invention (see for example Thompson, U.S. Pat. Nos. 5,902,880 and 6,146,886). The recombinant vectors capable of expressing the siNA molecules can be delivered as described above, and persist in target cells. Alternatively, viral vectors can be used that provide for transient expression of nucleic acid molecules. Such vectors can be repeatedly administered as necessary. Once expressed, the siNA molecule interacts with the target mRNA and generates an RNAi response. Delivery of siNA molecule expressing vectors can be systemic, such as by intravenous or intra-muscular administration, by administration to target cells ex-planted from a subject followed by reintroduction into the subject, or by any other means that would allow for introduction into the desired target cell (for a review see Couture et al., 1996,  TIG.,  12, 510). 
     In one aspect the invention features an expression vector comprising a nucleic acid sequence encoding at least one siNA molecule of the instant invention. The expression vector can encode one or both strands of a siNA duplex, or a single self-complementary strand that self hybridizes into a siNA duplex. The nucleic acid sequences encoding the siNA molecules of the instant invention can be operably linked in a manner that allows expression of the siNA molecule (see for example Paul et al., 2002,  Nature Biotechnology,  19, 505; Miyagishi and Taira, 2002,  Nature Biotechnology,  19, 497; Lee et al., 2002,  Nature Biotechnology,  19, 500; and Novina et al., 2002,  Nature Medicine, advance online publication doi: 10.1038/nm725). 
     In another aspect, the invention features an expression vector comprising: a) a transcription initiation region (e.g., eukaryotic pol I, II or III initiation region); b) a transcription termination region (e.g., eukaryotic pol I, II or III termination region); and c) a nucleic acid sequence encoding at least one of the siNA molecules of the instant invention, wherein said sequence is operably linked to said initiation region and said termination region in a manner that allows expression and/or delivery of the siNA molecule. The vector can optionally include an open reading frame (ORF) for a protein operably linked on the 5′ side or the 3′-side of the sequence encoding the siNA of the invention; and/or an intron (intervening sequences). 
     Transcription of the siNA molecule sequences can be driven from a promoter for eukaryotic RNA polymerase I (pol I), RNA polymerase II (pol II), or RNA polymerase III (pol III). Transcripts from pol II or pol III promoters are expressed at high levels in all cells; the levels of a given pol II promoter in a given cell type depends on the nature of the gene regulatory sequences (enhancers, silencers, etc.) present nearby. Prokaryotic RNA polymerase promoters are also used, providing that the prokaryotic RNA polymerase enzyme is expressed in the appropriate cells (Elroy-Stein and Moss, 1990,  Proc. Natl. Acad. Sci. USA,  87, 6743-7; Gao and Huang 1993,  Nucleic Acids Res.,  21, 2867-72; Lieber et al., 1993,  Methods Enzymol.,  217, 47-66; Zhou et al., 1990,  Mol. Cell. Biol.,  10, 4529-37). Several investigators have demonstrated that nucleic acid molecules expressed from such promoters can function in mammalian cells (e.g. Kashani-Sabet et al., 1992,  Antisense Res. Dev.,  2, 3-15; Ojwang et al., 1992,  Proc. Natl. Acad. Sci. USA,  89, 10802-6; Chen et al., 1992,  Nucleic Acids Res.,  20, 4581-9; Yu et al., 1993,  Proc. Natl. Acad. Sci. USA,  90, 6340-4; L&#39;Huillier et al., 1992,  EMBO J.,  11, 4411-8; Lisziewicz et al., 1993,  Proc. Natl. Acad. Sci. U.S.A.,  90, 8000-4; Thompson et al., 1995,  Nucleic Acids Res.,  23, 2259; Sullenger &amp; Cech, 1993,  Science,  262, 1566). More specifically, transcription units such as the ones derived from genes encoding U6 small nuclear (snRNA), transfer RNA (tRNA) and adenovirus VA RNA are useful in generating high concentrations of desired RNA molecules such as siNA in cells (Thompson et al., supra; Couture and Stinchcomb, 1996, supra; Noonberg et al., 1994,  Nucleic Acid Res.,  22, 2830; Noonberg et al., U.S. Pat. No. 5,624,803; Good et al., 1997,  Gene Ther.,  4, 45; Beigelman et al., International PCT Publication No. WO 96/18736. The above siNA transcription units can be incorporated into a variety of vectors for introduction into mammalian cells, including but not restricted to, plasmid DNA vectors, viral DNA vectors (such as adenovirus or adeno-associated virus vectors), or viral RNA vectors (such as retroviral or alphavirus vectors) (for a review see Couture and Stinchcomb, 1996, supra). 
     In another aspect the invention features an expression vector comprising a nucleic acid sequence encoding at least one of the siNA molecules of the invention in a manner that allows expression of that siNA molecule. The expression vector comprises in one embodiment; a) a transcription initiation region; b) a transcription termination region; and c) a nucleic acid sequence encoding at least one strand of the siNA molecule, wherein the sequence is operably linked to the initiation region and the termination region in a manner that allows expression and/or delivery of the siNA molecule. 
     In another embodiment the expression vector comprises: a) a transcription initiation region; b) a transcription termination region; c) an open reading frame; and d) a nucleic acid sequence encoding at least one strand of a siNA molecule, wherein the sequence is operably linked to the 3′-end of the open reading frame and wherein the sequence is operably linked to the initiation region, the open reading frame and the termination region in a manner that allows expression and/or delivery of the siNA molecule. In yet another embodiment, the expression vector comprises: a) a transcription initiation region; b) a transcription termination region; c) an intron; and d) a nucleic acid sequence encoding at least one siNA molecule, wherein the sequence is operably linked to the initiation region, the intron and the termination region in a manner which allows expression and/or delivery of the nucleic acid molecule. 
     In another embodiment, the expression vector comprises: a) a transcription initiation region; b) a transcription termination region; c) an intron; d) an open reading frame; and e) a nucleic acid sequence encoding at least one strand of a siNA molecule, wherein the sequence is operably linked to the 3′-end of the open reading frame and wherein the sequence is operably linked to the initiation region, the intron, the open reading frame and the termination region in a manner which allows expression and/or delivery of the siNA molecule. 
     Protein Tyrosine Phosphatase-1B Biology and Biochemistry 
     Protein tyrosine phosphorylation and dephosphorylation are important mechanisms in the regulation of signal transduction pathways that control the processes of cell growth, proliferation, and differentiation (Fantl, W. J., 1993, Annu. Rev. Biochem., 62, 453-481). Cooperative enzyme classes regulate protein tyrosine phosphorylation and dephosphorylation events. These broad classes of enzymes consist of the protein tyrosine kinases (PTKs) and protein tyrosine phosphatases (PTPs). PTKs and PTPs can exist as both receptor-type transmembrane proteins and as cytoplasmic protein enzymes. Receptor tyrosine kinases propagate signal transduction events via extracellular receptor-ligand interactions that result in the activation of the tyrosine kinase portion of the PTK in the cytoplasmic domain. Receptor-like transmembrane PTPs function through extracellular ligand binding that modulates dephosphorylation of intracellular phosphotyrosine proteins via cytoplasmic phosphatase domains. Cytoplasmic PTKs and PTPs exert enzymatic activity without receptor-mediated ligand interactions, however, phosphorylation can regulate the activity of these enzymes. 
     Protein tyrosine phosphatase 1B, a cytoplasmic PTP, was the first PTP to be isolated in homogeneous form (Tonics, N. K., 1988, J. Biol. Chem., 263, 6722-6730), characterized (Tonks, N. K., 1988, J. Biol. Chem., 263, 6731-6737), and sequenced (Charbonneau, H., 1989, Biochemistry, 86, 5252-5256). Cytoplasmic and receptor-like PTPs both share a catalytic domain characterized by eleven conserved amino acids containing cysteine and arginine residues that are critical for phosphatase activity (Streuli, M., 1990, EMBO, 9, 2399-2407). A cysteine residue at position 215 is responsible for the covalent attachment of phosphate to the enzyme (Guan, K., 1991, J. Biol. Chem., 266, 17026-17030). The crystal structure of human PTP-1B defined the phosphate binding site of the enzyme as a glycine rich cleft at the surface of the molecule with cysteine 215 positioned at the base of this cleft. The location of cysteine 215 and the shape of the cleft provide specificity of PTPase activity for tyrosine residues but not for serine or threonine residues (Barford, D., 1994, Science, 263, 1397-1404). 
     Receptor tyrosine kinase and protein tyrosine phosphatase localization plays a key role in the regulation of phosphotyrosine mediated signal transduction. PTP-1B activity and specificity against a panel of receptor tyrosine kinases demonstrated clear differences between substrates, suggesting that cellular compartmentalization is a determinant in defining the activity and function of the enzyme (Lammers, R., 1993, J. Biol. Chem., 268, 22456-22462). Experiments have indicated that PTP-1B is localized predominantly in the endoplasmic reticulum via its 35 amino acid carboxyterminal sequence. PTP-1B is also tightly associated with microsomal membranes with its catalytic phosphatase domain oriented towards the cytoplasm (Frangioni, J. V., 1992, Cell, 68, 545-560). 
     PTP-1B has been identified as a negative regulator of the insulin response. PTP-1B is widely expressed in insulin sensitive tissues (Goldstein, B. J., 1993, Receptor, 3, 1-15). Isolated PTP-1B dephosphorylates the insulin receptor in vitro (Tonics, N. K., 1988, J. Biol. Chem., 263, 6731-6737). PTP-1B dephosphorylation of multiple phosphotyrosine residues of the insulin receptor proceeds sequentially and with specificity for the three tyrosine residues that are critical for receptor autoactivation (Ramachandran, C., 1992, Biochemistry, 31, 4232-4238). In addition to insulin receptor dephosphorylation, PTP-1B also dephosphorylates the insulin related subtrate 1 (IRS-1), a principal substrate of the insulin receptor (Lammers, R., 1993, J. Biol. Chem., 268, 22456-22462). 
     Microinjection of PTP-1B into  Xenopus oocytes  results in the inhibition of insulin stimulated tyrosine phosphorylation of endogenous proteins, including the beta-subunit of the insulin and insulin-like growth factor receptor proteins. The resulting 3 to 5 fold increase over endogenous PTPase activity also blocks the activation of an S6 peptide kinase (Cicirelli, M. F., 1990, Proc, Natl. Acad. Sci., 87, 5514-5518). Inactivation of recombinant rat PTP-1B with antibody immunoprecipitation results in the dramatic increase in insulin stimulated DNA synthesis and phosphatidylinositol 3′-kinase activity. Insulin stimulated receptor autophosphorylation and insulin receptor substrate 1 tyrosine phosphorylation are increased dramatically as well through PTP-1B inhibition (Ahmad, F., 1995, J. Biol. Chem., 270, 20503-20508). 
     Increased PTP-1B expression correlates with insulin resistance in hyperglycemic cultured fibroblasts. In this study, desensitized insulin receptor function was observed via impaired insulin-induced autophosphorylation of the receptor. Treatment with insulin sensitivity normalizing thiazolidine derivatives resulted in the amelioration of the hyperglycemic insulin resistance via a normalization in PTP-1B expression (Maegawa, H., 1995, J. Biol. Chem., 270, 7724-7730). A murine model of insulin resistance with a knockout of the hetrerotrimeric GTP-binding protein subunit Gi-alpha-2 provides a type 2 diabetes phenotype that correlates with the increased expression of PTP-1B (Moxam, C. M., 1996, Nature, 379, 840-844). 
     PTP-1B interacts directly with the activated insulin receptor beta-subunit. An inactive homolog of PTP-1B was used to precipitate the activated insulin receptor in both purified receptor preparations and whole-cell lysates. Phosphorylation of the insulin receptor&#39;s triple tyrosine residues in the kinase domain is necessary for PTP-1B interaction. Furthermore, insulin stimulates tyrosine phosphorylation of PTP-1B (Seely, B. L., 1996, Diabetes, 45, 1379-1385). A similar study confirmed the direct interaction of PTP-1B with the insulin receptor beta-subunit as well as the required multiple phosphorylation sites within the receptor and PTP-1B (Bandyopadhyay, D., J. Biol. Chem., 272, 1639-1645). 
     Knockout mice lacking the PTP-1B gene (both homozygous PTP-1B −/−  and heterozygous PTP-1B +/− ) have been used to study the specific role of PTP-1B relating to insulin action in vivo. The resulting PTP-1B deficient mice were healthy and, in the fed state, had lower blood glucose and circulating insulin levels that were half that of their PTP-1B +/+  expressing littermates. These PTP-1B deficient mice demonstrated enhanced insulin sensitivity in glucose and insulin tolerance tests. At the physiological level, the PTP-1B deficient mice showed increased phosphorylation of the insulin receptor after insulin administration. When fed a high fat diet, the PTP-1B deficient mice were resistant to weight gain and remained insulin sensitive as opposed to normal PTP-1B expressing mice, who rapidly gained weight and become insulin resistant (Elchebly, M., 1999, Science, 283, 1544-1548). As such, modulation of PTP-1B expression could be used to regulate autophosphorylation of the insulin receptor and increase insulin sensitivity in vivo. This modulation could prove beneficial in the treatment of insulin related disease states. 
     In light of the above findings, particular disease states that involve PTP-1B expression include but are not limited to:
     1. Diabetes: Both type 1 and type 2 diabetes can be treated by modulation of PTP-1B expression. Type 2 diabetes correlates to desensitized insulin receptor function (White et al., 1994). Disruption of the PTP-1B dephosphorylation of the insulin receptor in vivo manifests in insulin sensitivity and increased insulin receptor autophosphorylation (Elchebly et al., 1999). Insulin dependant diabetes, type 1, can respond to PTP-1B modulation through increased insulin sensitivity.   2. Obesity: Elchebly et al., 1999, demonstrated that PTP-1B deficient mice were resistant to weight gain when fed a high fat diet compared to normal PTP-1B expressing mice. This finding suggests that PTP-1B modulation can be beneficial in the treatment of obesity. Ahmad et al., 1997, Metab. Clin. Exp., 46, 1140-1145, describe reduced PTPs in adipose tissue and improved insulin sensitivity in obese subjects following weight loss.   

     The human genome is thought to contain up to 100 PTPases, each varying slightly in chemistry but vastly in function. Compounds designed to inhibit PTP-1B activity specifically by covalent binding to or modification of PTP-1B have the potential for multiple side effects. Conventional drug substances that will potently suppress PTP-1B activity with few or no side effects from interaction with other PTPs are difficult to envision. A more attractive approach to PTP-1B modulation would involve the specific regulation of PTP-1B expression with nucleic acid technologies such as siRNA mediated RNAi. 
     Based upon the current understanding of PTP-1B, the modulation of PTP-1B and related genes is instrumental in the development of new therapeutics for obesity and diabetes. As such, modulation of PTP-1B using small interfering nucleic acid (siNA) mediated RNAi represents a novel approach to the treatment and study of diseases and conditions related PTP-1B activity and/or gene expression. 
     EXAMPLES 
     The following are non-limiting examples showing the selection, isolation, synthesis and activity of nucleic acids of the instant invention. 
     Example 1 
     Tandem Synthesis of siNA Constructs 
     Exemplary siNA molecules of the invention are synthesized in tandem using a cleavable linker, for example, a succinyl-based linker. Tandem synthesis as described herein is followed by a one-step purification process that provides RNAi molecules in high yield. This approach is highly amenable to siNA synthesis in support of high throughput RNAi screening, and can be readily adapted to multi-column or multi-well synthesis platforms. 
     After completing a tandem synthesis of a siNA oligo and its complement in which the 5′-terminal dimethoxytrityl (5′-O-DMT) group remains intact (trityl on synthesis), the oligonucleotides are deprotected as described above. Following deprotection, the siNA sequence strands are allowed to spontaneously hybridize. This hybridization yields a duplex in which one strand has retained the 5′-O-DMT group while the complementary strand comprises a terminal 5′-hydroxyl. The newly formed duplex behaves as a single molecule during routine solid-phase extraction purification (Trityl-On purification) even though only one molecule has a dimethoxytrityl group. Because the strands form a stable duplex, this dimethoxytrityl group (or an equivalent group, such as other trityl groups or other hydrophobic moieties) is all that is required to purify the pair of oligos, for example, by using a C18 cartridge. 
     Standard phosphoramidite synthesis chemistry is used up to the point of introducing a tandem linker, such as an inverted deoxy abasic succinate or glyceryl succinate linker (see  FIG. 1 ) or an equivalent cleavable linker. A non-limiting example of linker coupling conditions that can be used includes a hindered base such as diisopropylethylamine (DIPA) and/or DMAP in the presence of an activator reagent such as Bromotripyrrolidinophosphoniumhexaflurorophosphate (PyBrOP). After the linker is coupled, standard synthesis chemistry is utilized to complete synthesis of the second sequence leaving the terminal the 5′-O-DMT intact. Following synthesis, the resulting oligonucleotide is deprotected according to the procedures described herein and quenched with a suitable buffer, for example with 50 mM NaOAc or 1.5M NH 4 H 2 CO 3 . 
     Purification of the siNA duplex can be readily accomplished using solid phase extraction, for example, using a Waters C18 SepPak 1 g cartridge conditioned with 1 column volume (CV) of acetonitrile, 2 CV H2O, and 2 CV 50 mM NaOAc. The sample is loaded and then washed with 1 CV H2O or 50 mM NaOAc. Failure sequences are eluted with 1 CV 14% ACN (Aqueous with 50 mM NaOAc and 50 mM NaCl). The column is then washed, for example with 1 CV H2O followed by on-column detritylation, for example by passing 1 CV of 1% aqueous trifluoroacetic acid (TFA) over the column, then adding a second CV of 1% aqueous TFA to the column and allowing to stand for approximately 10 minutes. The remaining TFA solution is removed and the column washed with H20 followed by 1 CV 1M NaCl and additional H2O. The siNA duplex product is then eluted, for example, using 1 CV 20% aqueous CAN. 
       FIG. 2  provides an example of MALDI-TOF mass spectrometry analysis of a purified siNA construct in which each peak corresponds to the calculated mass of an individual siNA strand of the siNA duplex. The same purified siNA provides three peaks when analyzed by capillary gel electrophoresis (CGE), one peak presumably corresponding to the duplex siNA, and two peaks presumably corresponding to the separate siNA sequence strands. Ion exchange HPLC analysis of the same siNA contract only shows a single peak. Testing of the purified siNA construct using a luciferase reporter assay described below demonstrated the same RNAi activity compared to siNA constructs generated from separately synthesized oligonucleotide sequence strands. 
     Example 2 
     Identification of Potential siNA Target Sites in any RNA Sequence 
     The sequence of an RNA target of interest, such as a viral or human mRNA transcript, is screened for target sites, for example by using a computer folding algorithm. In a non-limiting example, the sequence of a gene or RNA gene transcript derived from a database, such as Genbank, is used to generate siNA targets having complementarity to the target. Such sequences can be obtained from a database, or can be determined experimentally as known in the art. Target sites that are known, for example, those target sites determined to be effective target sites based on studies with other nucleic acid molecules, for example ribozymes or antisense, or those targets known to be associated with a disease, trait, or condition such as those sites containing mutations or deletions, can be used to design siNA molecules targeting those sites. Various parameters can be used to determine which sites are the most suitable target sites within the target RNA sequence. These parameters include but are not limited to secondary or tertiary RNA structure, the nucleotide base composition of the target sequence, the degree of homology between various regions of the target sequence, or the relative position of the target sequence within the RNA transcript. Based on these determinations, any number of target sites within the RNA transcript can be chosen to screen siNA molecules for efficacy, for example by using in vitro RNA cleavage assays, cell culture, or animal models. In a non-limiting example, anywhere from 1 to 1000 target sites are chosen within the transcript based on the size of the siNA construct to be used. High throughput screening assays can be developed for screening siNA molecules using methods known in the art, such as with multi-well or multi-plate assays to determine efficient reduction in target gene expression. 
     Example 3 
     Selection of siNA Molecule Target Sites in a RNA 
     The following non-limiting steps can be used to carry out the selection of siNAs targeting a given gene sequence or transcript.
     1. The target sequence is parsed in silico into a list of all fragments or subsequences of a particular length, for example 23 nucleotide fragments, contained within the target sequence. This step is typically carried out using a custom Perl script, but commercial sequence analysis programs such as Oligo, MacVector, or the GCG Wisconsin Package can be employed as well.   2. In some instances the siNAs correspond to more than one target sequence; such would be the case for example in targeting different transcripts of the same gene, targeting different transcripts of more than one gene, or for targeting both the human gene and an animal homolog. In this case, a subsequence list of a particular length is generated for each of the targets, and then the lists are compared to find matching sequences in each list. The subsequences are then ranked according to the number of target sequences that contain the given subsequence; the goal is to find subsequences that are present in most or all of the target sequences. Alternately, the ranking can identify subsequences that are unique to a target sequence, such as a mutant target sequence. Such an approach would enable the use of siNA to target specifically the mutant sequence and not effect the expression of the normal sequence.   3. In some instances the siNA subsequences are absent in one or more sequences while present in the desired target sequence; such would be the case if the siNA targets a gene with a paralogous family member that is to remain untargeted. As in case 2 above, a subsequence list of a particular length is generated for each of the targets, and then the lists are compared to find sequences that are present in the target gene but are absent in the untargeted paralog.   4. The ranked siNA subsequences can be further analyzed and ranked according to GC content. A preference can be given to sites containing 30-70% GC, with a further preference to sites containing 40-60% GC.   5. The ranked siNA subsequences can be further analyzed and ranked according to self-folding and internal hairpins. Weaker internal folds are preferred; strong hairpin structures are to be avoided.   6. The ranked siNA subsequences can be further analyzed and ranked according to whether they have runs of GGG or CCC in the sequence. GGG (or even more Gs) in either strand can make oligonucleotide synthesis problematic and can potentially interfere with RNAi activity, so it is avoided whenever better sequences are available. CCC is searched in the target strand because that will place GGG in the antisense strand.   7. The ranked siNA subsequences can be further analyzed and ranked according to whether they have the dinucleotide UU (uridine dinucleotide) on the 3′-end of the sequence, and/or AA on the 5′-end of the sequence (to yield 3′ UU on the antisense sequence). These sequences allow one to design siNA molecules with terminal TT thymidine dinucleotides.   8. Four or five target sites are chosen from the ranked list of subsequences as described above. For example, in subsequences having 23 nucleotides, the right 21 nucleotides of each chosen 23-mer subsequence are then designed and synthesized for the upper (sense) strand of the siNA duplex, while the reverse complement of the left 21 nucleotides of each chosen 23-mer subsequence are then designed and synthesized for the lower (antisense) strand of the siNA duplex (see Tables II and III). If terminal TT residues are desired for the sequence (as described in paragraph 7), then the two 3′ terminal nucleotides of both the sense and antisense strands are replaced by TT prior to synthesizing the oligos.   9. The siNA molecules are screened in an in vitro, cell culture or animal model system to identify the most active siNA molecule or the most preferred target site within the target RNA sequence.   10. Other design considerations can be used when selecting target nucleic acid sequences, see, for example, Reynolds et al., 2004,  Nature Biotechnology Advanced Online Publication,  1 Feb. 2004, doi:10.1038/nbt936 and Ui-Tei et al., 2004, Nucleic Acids Research, 32, doi:10.1093/nar/gkh247.   

     In an alternate approach, a pool of siNA constructs specific to a PTP-1B target sequence is used to screen for target sites in cells expressing PTP-1B RNA, such as cultured Jurkat, HeLa, A549 or 293T cells. The general strategy used in this approach is shown in  FIG. 9 . A non-limiting example of such is a pool comprising sequences having any of SEQ ID NOS 1-755. Cells expressing PTP-1B are transfected with the pool of siNA constructs and cells that demonstrate a phenotype associated with PTP-1B inhibition are sorted. The pool of siNA constructs can be expressed from transcription cassettes inserted into appropriate vectors (see for example  FIG. 7  and  FIG. 8 ). The siNA from cells demonstrating a positive phenotypic change (e.g., decreased proliferation, decreased PTP-1B mRNA levels or decreased PTP-1B protein expression), are sequenced to determine the most suitable target site(s) within the target PTP-1B RNA sequence. 
     Example 4 
     PTP-1B Targeted siNA Design 
     siNA target sites were chosen by analyzing sequences of the PTP-1B RNA target and optionally prioritizing the target sites on the basis of folding (structure of any given sequence analyzed to determine siNA accessibility to the target), by using a library of siNA molecules as described in Example 3, or alternately by using an in vitro siNA system as described in Example 6 herein. siNA molecules were designed that could bind each target and are optionally individually analyzed by computer folding to assess whether the siNA molecule can interact with the target sequence. Varying the length of the siNA molecules can be chosen to optimize activity. Generally, a sufficient number of complementary nucleotide bases are chosen to bind to, or otherwise interact with, the target RNA, but the degree of complementarity can be modulated to accommodate siNA duplexes or varying length or base composition. By using such methodologies, siNA molecules can be designed to target sites within any known RNA sequence, for example those RNA sequences corresponding to the any gene transcript. 
     Chemically modified siNA constructs are designed to provide nuclease stability for systemic administration in vivo and/or improved pharmacokinetic, localization, and delivery properties while preserving the ability to mediate RNAi activity. Chemical modifications as described herein are introduced synthetically using synthetic methods described herein and those generally known in the art. The synthetic siNA constructs are then assayed for nuclease stability in serum and/or cellular/tissue extracts (e.g. liver extracts). The synthetic siNA constructs are also tested in parallel for RNAi activity using an appropriate assay, such as a luciferase reporter assay as described herein or another suitable assay that can quantity RNAi activity. Synthetic siNA constructs that possess both nuclease stability and RNAi activity can be further modified and re-evaluated in stability and activity assays. The chemical modifications of the stabilized active siNA constructs can then be applied to any siNA sequence targeting any chosen RNA and used, for example, in target screening assays to pick lead siNA compounds for therapeutic development (see for example  FIG. 11 ). 
     Example 5 
     Chemical Synthesis and Purification of siNA 
     siNA molecules can be designed to interact with various sites in the RNA message, for example, target sequences within the RNA sequences described herein. The sequence of one strand of the siNA molecule(s) is complementary to the target site sequences described above. The siNA molecules can be chemically synthesized using methods described herein. Inactive siNA molecules that are used as control sequences can be synthesized by scrambling the sequence of the siNA molecules such that it is not complementary to the target sequence. Generally, siNA constructs can by synthesized using solid phase oligonucleotide synthesis methods as described herein (see for example Usman et al., U.S. Pat. Nos. 5,804,683; 5,831,071; 5,998,203; 6,117,657; 6,353,098; 6,362,323; 6,437,117; 6,469,158; Scaringe et al., U.S. Pat. Nos. 6,111,086; 6,008,400; 6,111,086 all incorporated by reference herein in their entirety). 
     In a non-limiting example, RNA oligonucleotides are synthesized in a stepwise fashion using the phosphoramidite chemistry as is known in the art. Standard phosphoramidite chemistry involves the use of nucleosides comprising any of 5′-O-dimethoxytrityl, 2′-O-tert-butyldimethylsilyl, 3′-O-2-Cyanoethyl N,N-diisopropylphosphoroamidite groups, and exocyclic amine protecting groups (e.g. N6-benzoyl adenosine, N4 acetyl cytidine, and N2-isobutyryl guanosine). Alternately, 2′-O-Silyl Ethers can be used in conjunction with acid-labile 2′-O-orthoester protecting groups in the synthesis of RNA as described by Scaringe supra. Differing 2′ chemistries can require different protecting groups, for example 2′-deoxy-2′-amino nucleosides can utilize N-phthaloyl protection as described by Usman et al., U.S. Pat. No. 5,631,360, incorporated by reference herein in its entirety). 
     During solid phase synthesis, each nucleotide is added sequentially (3′- to 5′-direction) to the solid support-bound oligonucleotide. The first nucleoside at the 3′-end of the chain is covalently attached to a solid support (e.g., controlled pore glass or polystyrene) using various linkers. The nucleotide precursor, a ribonucleoside phosphoramidite, and activator are combined resulting in the coupling of the second nucleoside phosphoramidite onto the 5′-end of the first nucleoside. The support is then washed and any unreacted 5′-hydroxyl groups are capped with a capping reagent such as acetic anhydride to yield inactive 5′-acetyl moieties. The trivalent phosphorus linkage is then oxidized to a more stable phosphate linkage. At the end of the nucleotide addition cycle, the 5′-O-protecting group is cleaved under suitable conditions (e.g., acidic conditions for trityl-based groups and Fluoride for silyl-based groups). The cycle is repeated for each subsequent nucleotide. 
     Modification of synthesis conditions can be used to optimize coupling efficiency, for example by using differing coupling times, differing reagent/phosphoramidite concentrations, differing contact times, differing solid supports and solid support linker chemistries depending on the particular chemical composition of the siNA to be synthesized. Deprotection and purification of the siNA can be performed as is generally described in Usman et al., U.S. Pat. No. 5,831,071, U.S. Pat. No. 6,353,098, U.S. Pat. No. 6,437,117, and Bellon et al., U.S. Pat. No. 6,054,576, U.S. Pat. No. 6,162,909, U.S. Pat. No. 6,303,773, or Scaringe supra, incorporated by reference herein in their entireties. Additionally, deprotection conditions can be modified to provide the best possible yield and purity of siNA constructs. For example, applicant has observed that oligonucleotides comprising 2′-deoxy-2′-fluoro nucleotides can degrade under inappropriate deprotection conditions. Such oligonucleotides are deprotected using aqueous methylamine at about 35° C. for 30 minutes. If the 2′-deoxy-2′-fluoro containing oligonucleotide also comprises ribonucleotides, after deprotection with aqueous methylamine at about 35° C. for 30 minutes, TEA-HF is added and the reaction maintained at about 65° C. for an additional 15 minutes. 
     Example 6 
     RNAi In Vitro Assay to Assess siNA Activity 
     An in vitro assay that recapitulates RNAi in a cell-free system is used to evaluate siNA constructs targeting PTP-1B RNA targets. The assay comprises the system described by Tuschl et al., 1999,  Genes and Development,  13, 3191-3197 and Zamore et al., 2000,  Cell,  101, 25-33 adapted for use with PTP-1B target RNA. A  Drosophila  extract derived from syncytial blastoderm is used to reconstitute RNAi activity in vitro. Target RNA is generated via in vitro transcription from an appropriate PTP-1B expressing plasmid using T7 RNA polymerase or via chemical synthesis as described herein. Sense and antisense siNA strands (for example 20 uM each) are annealed by incubation in buffer (such as 100 mM potassium acetate, 30 mM HEPES-KOH, pH 7.4, 2 mM magnesium acetate) for 1 minute at 90° C. followed by 1 hour at 37° C., then diluted in lysis buffer (for example 100 mM potassium acetate, 30 mM HEPES-KOH at pH 7.4, 2 mM magnesium acetate). Annealing can be monitored by gel electrophoresis on an agarose gel in TBE buffer and stained with ethidium bromide. The  Drosophila  lysate is prepared using zero to two-hour-old embryos from Oregon R flies collected on yeasted molasses agar that are dechorionated and lysed. The lysate is centrifuged and the supernatant isolated. The assay comprises a reaction mixture containing 50% lysate [vol/vol], RNA (10-50 pM final concentration), and 10% [vol/vol] lysis buffer containing siNA (10 nM final concentration). The reaction mixture also contains 10 mM creatine phosphate, 10 ug/ml creatine phosphokinase, 100 um GTP, 100 uM UTP, 100 uM CTP, 500 uM ATP, 5 mM DTT, 0.1 U/uL RNasin (Promega), and 100 uM of each amino acid. The final concentration of potassium acetate is adjusted to 100 mM. The reactions are pre-assembled on ice and preincubated at 25° C. for 10 minutes before adding RNA, then incubated at 25° C. for an additional 60 minutes. Reactions are quenched with 4 volumes of 1.25× Passive Lysis Buffer (Promega). Target RNA cleavage is assayed by RT-PCR analysis or other methods known in the art and are compared to control reactions in which siNA is omitted from the reaction. 
     Alternately, internally-labeled target RNA for the assay is prepared by in vitro transcription in the presence of [alpha- 32 P] CTP, passed over a G50 Sephadex column by spin chromatography and used as target RNA without further purification. Optionally, target RNA is 5′- 32 P-end labeled using T4 polynucleotide kinase enzyme. Assays are performed as described above and target RNA and the specific RNA cleavage products generated by RNAi are visualized on an autoradiograph of a gel. The percentage of cleavage is determined by PHOSPHOR IMAGER® (autoradiography) quantitation of bands representing intact control RNA or RNA from control reactions without siNA and the cleavage products generated by the assay. 
     In one embodiment, this assay is used to determine target sites in the PTP-1B RNA target for siNA mediated RNAi cleavage, wherein a plurality of siNA constructs are screened for RNAi mediated cleavage of the PTP-1B RNA target, for example, by analyzing the assay reaction by electrophoresis of labeled target RNA, or by northern blotting, as well as by other methodology well known in the art. 
     Example 7 
     Nucleic Acid Inhibition of PTP-1B Target RNA In Vivo 
     siNA molecules targeted to the huma PTP-1B RNA are designed and synthesized as described above. These nucleic acid molecules can be tested for cleavage activity in vivo, for example, using the following procedure. The target sequences and the nucleotide location within the PTP-1B RNA are given in Table II and III. 
     Two formats are used to test the efficacy of siNAs targeting PTP-1B. First, the reagents are tested in cell culture using, for example, Jurkat, HeLa, A549 or 293T cells, to determine the extent of RNA and protein inhibition. siNA reagents (e.g.; see Tables II and III) are selected against the PTP-1B target as described herein. RNA inhibition is measured after delivery of these reagents by a suitable transfection agent to, for example, Jurkat, HeLa, A549 or 293T cells. Relative amounts of target RNA are measured versus actin using real-time PCR monitoring of amplification (eg., ABI 7700 TAQMAN®). A comparison is made to a mixture of oligonucleotide sequences made to unrelated targets or to a randomized siNA control with the same overall length and chemistry, but randomly substituted at each position. Primary and secondary lead reagents are chosen for the target and optimization performed. After an optimal transfection agent concentration is chosen, a RNA time-course of inhibition is performed with the lead siNA molecule. In addition, a cell-plating format can be used to determine RNA inhibition. 
     Delivery of siNA to Cells 
     Cells (e.g., Jurkat, HeLa, A549 or 293T cells) are seeded, for example, at 1×10 5  cells per well of a six-well dish in EGM-2 (BioWhittaker) the day before transfection. siNA (final concentration, for example 20 nM) and cationic lipid (e.g., final concentration 2 μg/ml) are complexed in EGM basal media (Biowhittaker) at 37° C. for 30 minutes in polystyrene tubes. Following vortexing, the complexed siNA is added to each well and incubated for the times indicated. For initial optimization experiments, cells are seeded, for example, at 1×10 3  in 96 well plates and siNA complex added as described. Efficiency of delivery of siNA to cells is determined using a fluorescent siNA complexed with lipid. Cells in 6-well dishes are incubated with siNA for 24 hours, rinsed with PBS and fixed in 2% paraformaldehyde for 15 minutes at room temperature. Uptake of siNA is visualized using a fluorescent microscope. 
     TAQMAN® (Real-Time PCR Monitoring of Amplification) and Lightcycler Quantification of mRNA 
     Total RNA is prepared from cells following siNA delivery, for example, using Qiagen RNA purification kits for 6-well or Rneasy extraction kits for 96-well assays. For TAQMAN® analysis (real-time PCR monitoring of amplification), dual-labeled probes are synthesized with the reporter dye, FAM or JOE, covalently linked at the 5′-end and the quencher dye TAMRA conjugated to the 3′-end. One-step RT-PCR amplifications are performed on, for example, an ABI PRISM 7700 Sequence Detector using 50 μl reactions consisting of 10 μl total RNA, 100 nM forward primer, 900 nM reverse primer, 100 nM probe, 1× TaqMan PCR reaction buffer (PE-Applied Biosystems), 5.5 mM MgCl 2 , 300 μM each dATP, dCTP, dGTP, and dTTP, 10 U RNase Inhibitor (Promega), 1.25 U AMPLTTAQ GOLD® (DNA polymerase) (PE-Applied Biosystems) and 10 U M-MLV Reverse Transcriptase (Promega). The thermal cycling conditions can consist of 30 minutes at 48° C., 10 minutes at 95° C., followed by 40 cycles of 15 seconds at 95° C. and 1 minute at 60° C. Quantitation of mRNA levels is determined relative to standards generated from serially diluted total cellular RNA (300, 100, 33, 11 ng/reaction) and normalizing to β-actin or GAPDH mRNA in parallel TAQMAN® reactions (real-time PCR monitoring of amplification). For each gene of interest an upper and lower primer and a fluorescently labeled probe are designed. Real time incorporation of SYBR Green 1 dye into a specific PCR product can be measured in glass capillary tubes using a lightcyler. A standard curve is generated for each primer pair using control cRNA. Values are represented as relative expression to GAPDH in each sample. 
     Western Blotting 
     Nuclear extracts can be prepared using a standard micro preparation technique (see for example Andrews and Faller, 1991,  Nucleic Acids Research,  19, 2499). Protein extracts from supernatants are prepared, for example using TCA precipitation. An equal volume of 20% TCA is added to the cell supernatant, incubated on ice for 1 hour and pelleted by centrifugation for 5 minutes. Pellets are washed in acetone, dried and resuspended in water. Cellular protein extracts are run on a 10% Bis-Tris NuPage (nuclear extracts) or 4-12% Tris-Glycine (supernatant extracts) polyacrylamide gel and transferred onto nitro-cellulose membranes. Non-specific binding can be blocked by incubation, for example, with 5% non-fat milk for 1 hour followed by primary antibody for 16 hour at 4° C. Following washes, the secondary antibody is applied, for example (1:10,000 dilution) for 1 hour at room temperature and the signal detected with SuperSignal reagent (Pierce). 
     Example 8 
     Models Useful to Evaluate the Down-Regulation of PTP-1B Gene Expression 
     Cell Culture 
     There are numerous cell culture systems that can be used to analyze reduction of PTP-1B levels either directly or indirectly by measuring downstream effects. For example, cultured human kidney fibroblast cells (e.g., 293 cells) can be used in cell culture experiments to assess the efficacy of nucleic acid molecules of the invention. As such, 293 cells treated with nucleic acid molecules of the invention (e.g., siNA) targeting PTP-1B RNA would be expected to have decreased PTP-1B expression capacity compared to matched control nucleic acid molecules having a scrambled or inactive sequence. In a non-limiting example, human kidney fibroblast 293 cells are cultured and PTP-1B expression is quantified, for example by time-resolved immuno fluorometric assay. PTP-1B messenger-RNA expression is quantitated with RT-PCR in cultured 293s. Untreated cells are compared to cells treated with siNA molecules transfected with a suitable reagent, for example a cationic lipid such as lipofectamine, and PTP-1B protein and RNA levels are quantitated. Dose response assays are then performed to establish dose dependent inhibition of PTP-1B expression. Other non-limiting examples of cell culture systems that can be used to assay nucleic acid molecules of the instant invention include: Maegawa et al., 1995, J. Biol. Chem., 270, 7724-7730, who describe a tissue culture model in which Rat 1 fibroblasts expressing human insulin receptors can be used to model hyperglycemia induced insulin resistance. Maegawa et al. also describe assays to measure PTPase activity using labeled phosphorylated insulin receptors and by immunoenzymatic techniques. Moxham et al., 1996, Nature, 379, 840-844, describe a murine tissue culture model employing Gi-alpha-2 deficiency to study hyperinsulinaemia, impaired glucose tolerance and resistance to insulin in vivo. Assays for PTPase activity and and tyrosine phosphorylation of insulin-receptor substrate 1 are known in the art. For example, Wang et al., 1999, Biochim. Biophys. Acta, 1431, 14-23, describe fluorescein monophosphates as fluorogenic substrates for PTPs which can be used to study PTPase modulation. The use of such fluorogenic PTP-1B substrates can be used to develop a high throughput screening assay for siRNA-based inhibition of PTP-1B in vivo. 
     In several cell culture systems, cationic lipids have been shown to enhance the bioavailability of oligonucleotides to cells in culture (Bennet, et al., 1992,  Mol. Pharmacology,  41, 1023-1033). In one embodiment, siNA molecules of the invention are complexed with cationic lipids for cell culture experiments. siNA and cationic lipid mixtures are prepared in serum-free DMEM immediately prior to addition to the cells. DMEM plus additives are warmed to room temperature (about 20-25° C.) and cationic lipid is added to the final desired concentration and the solution is vortexed briefly. siNA molecules are added to the final desired concentration and the solution is again vortexed briefly and incubated for 10 minutes at room temperature. In dose response experiments, the RNA/lipid complex is serially diluted into DMEM following the 10 minute incubation. 
     Animal Models 
     Evaluating the efficacy of anti-PTP-1B agents in animal models is an important prerequisite to human clinical trials. Obesity and type 2 diabetes are the most prevalent and serious metabolic diseases in that they affect more than 50% of adults in the USA. These conditions are associated with a chronic inflammatory response characterized by abnormal inflammatory cytokine production, increased acute-phase reactants and other stress-induced molecules. Many of these alterations seem to be initiated and to reside within adipose tissue. Elevated production of tumour necrosis factor (TNF)-alpha by adipose tissue decreases sensitivity to insulin and has been detected in several experimental obesity models and obese humans. Free fatty acids (FFAs) are also implicated in the etiology of obesity-induced insulin resistance and diabetes. Investigation has shown that obesity is associated with alterations in stress-activated and inflammatory responses through the tyrosine kinase pathway and that protein kinases are causally linked to aberrant metabolic control in this state. Hirosumi et al., 2002,  Nature,  420, 333-336. Hirosumi et al., describe dietary and genetic (ob/ob) mouse models of obesity useful in evaluating PTP-1B gene expression. Other models include those described by Khandelwal et al., 1995,  Molecular and Cellular Biochemistry,  153, 87-94, who describe four different animal models for studying insulin dependent and insulin resistant diabetes mellitus. These models were used to study the effect of vanadate, an insulin mimetic and PTPase inhibitor, on the insulin-stimulated phosphorylation of the insulin receptor and its tyrosine kinase activity. Elchebly et al., 1999, Science, 283, 1544-1548, describe a murine PTP-1B knockout model in which insulin sensitivity and fuel metabolism are studied. The resulting PTP-1B deficient mice (both homozygous PTP-1B −/−  and heterozygous PTP-1B +/− ) were healthy and, in the fed state, had lower blood glucose and circulating insulin levels that were one-half that of their PTP-1B +/+  expressing littermates. These PTP-1B deficient mice demonstrated enhanced insulin sensitivity in glucose and insulin tolerance tests. At the physiological level, the PTP-1B deficient mice showed increased phosphorylation of the insulin receptor after insulin administration. When fed a high fat diet, the PTP-1B deficient mice were resistant to weight gain and remained insulin sensitive as opposed to normal PTP-1B expressing mice, who rapidly gained weight and become insulin resistant. 
     Such transgenic mice are useful as models for obesity and insulin resistance and can be used to identify nucleic acid molecules of the invention that modulate PTP-1B gene expression and gene function toward therapeutic use in treating obesity and insulin resistance (e.g. type I and II diabetes). 
     Example 9 
     RNAi Mediated Inhibition of PTP-1B Expression 
     In Vitro siNA Mediated Inhibition of PTP-1B RNA 
     siNA constructs (Table III) are tested for efficacy in reducing PTP-1B RNA expression in, for example, A549 cells. Cells are plated approximately 24 hours before transfection in 96-well plates at 5,000-7,500 cells/well, 100 such that at the time of transfection cells are 70-90% confluent. For transfection, annealed siNAs are mixed with the transfection reagent (Lipofectamine 2000, Invitrogen) in a volume of 50 μl/well and incubated for 20 minutes at room temperature. The siNA transfection mixtures are added to cells to give a final siNA concentration of 25 nM in a volume of 150 μl. Each siNA transfection mixture is added to 3 wells for triplicate siNA treatments. Cells are incubated at 37° for 24 hours in the continued presence of the siNA transfection mixture. At 24 hours, RNA is prepared from each well of treated cells. The supernatants with the transfection mixtures are first removed and discarded, then the cells are lysed and RNA prepared from each well. Target gene expression following treatment is evaluated by RT-PCR for the target gene and for a control gene (36B4, an RNA polymerase subunit) for normalization. The triplicate data is averaged and the standard deviations determined for each treatment. Normalized data are graphed and the percent reduction of target mRNA by active siNAs in comparison to their respective inverted control siNAs is determined. 
     Results of a non-limiting example are shown in  FIG. 29 . A siNA construct comprising ribonucleotides and 3′-terminal dithymidine caps (compound number 31018/31094) was compared to a chemically modified siNA construct comprising 2′-deoxy-2′-fluoro pyrimidine nucleotides and purine ribonucleotides in which the sense strand of the siNA is further modified with 5′ and 3′-terminal inverted deoxyabasic caps and the antisense strand comprises a 3′-terminal phosphorothioate internucleotide linkage (compound number 31306/31307), which was also compared to a matched chemistry inverted control (compound number 31318/31319). In addition, the siNA constructs were also compared to untreated cells, cells transfected with lipid and scrambled siNA constructs (Scram1 and Scram2), and cells transfected with lipid alone (transfection control). As shown in the figure, both siNA constructs show significant reduction of PTP-1B RNA expression compared with appropriate controls. 
     A follow up dose response study was used to determine the in vitro potency of stabilized siNAs targeting PTP-1B site 761. The mouse hepatoma cell line Hepa1-6 was plated at 7,500 cells/well. The cells were transfected with Lipofectamine2000 at a concentration equal to 0.35 ul/well. Dose response curves were carried out using PTP-1B site 761 stab 4/8 and 7/8 siNA constructs at an initial concentration of 50 nM diluted two-fold down to 781 pM. After a 24 hour incubation, the cells were harvested, total RNA isolated, and the level of PTP-1B mRNA was determined by real time RT_PCR using PTP-1B specific primers and probe. Results are shown in  FIG. 30 . An IC50 of ˜2 nM was observed with both the 4/8 and 7/8 stabilized siNAs targeting PTP-1B site 761. 
     In vivo siNA Mediated Inhibition of PTP-1B RNA 
     This study was designed to evaluate siNA-mediated reductions of PTP-1B mRNA levels in the livers of mice treated systemically with PTP-1B specific siNA constructs both with and without cholesterol conjugation. RNA oligonucleotides were synthesized using Phrarmacia AKTA synthesizers and standard phosphoramidite chemistry. The 5′-end of the sense strand is coupled to a cholesterol phosphoramidite ( FIG. 31 ) to obtain the cholesterol conjugate. Modified RNA oligonucleotides were deprotected deprotected using aqueous 40% methylamine at 350 C for 45 min, followed by triethylamine.3HF at 65 C for 1 hour. The crude material was purified by anion exchange chromatography and the full length fractions were pooled and the two strands subsequently annealed. The annealed product was desalted and then lyophilized to obtain the duplex siNA. 
     CD-1 mice (N=5 per group) were dosed intravenously with unformulated siNA at 30 mg/kg TID, and the cholesterol conjugated siNAs at 30 mg/kg/BID for three days. Sixteen hours post the last dose, animals were sacrificed. Total RNA was isolated from liver, and RT-PCR was performed with PTP-1B specific primers and probe. No reductions in liver PTP-1B mRNA was observed in the unformulated active siNA treated groups as compared to the irrelevant control or PBS treated groups. However, the groups treated with the cholesterol conjugated siRNAs in either Stab 4/8 (compound 39209/39709) or 7/8 (compound 39305/39709) chemistry (Table III) showed an ˜70% reduction in liver PTP-1B mRNA levels as compared to the corresponding irrelevant control or PBS treated groups ( FIG. 32 ). 
     Example 10 
     Indications 
     The present body of knowledge in PTP-1B research indicates the need for methods and compounds that can regulate PTP-1B gene product expression for research, diagnostic, and therapeutic use. Particular degenerative and disease states that can be associated with PTP-1B expression modulation include but are not limited to:
         Diabetes: Both type 1 and type 2 diabetes may be treated by modulation of PTP-1B expression. Type 2 diabetes correlates to desensitized insulin receptor function (White et al., 1994). Disruption of the PTP-1B dephosphorylation of the insulin receptor in vivo manifests in insulin sensitivity and increased insulin receptor autophosphorylation (Elchcbly et al., 1999). Insulin dependant diabetes, type 1, may respond to PTP-1B modulation through increased insulin sensitivity.   Obesity: Elchebly et al., 1999, demonstrated that PTP-1B deficient mice were resistant to weight gain when fed a high fat diet compared to normal PTP-1B expressing mice. This finding suggests that PTP-1B modulation may be beneficial in the treatment of obesity. Ahmad et al., 1997, Metab. Clin. Exp., 46, 1140-1145, describe reduced PTPs in adipose tissue and improved insulin sensitivity in obese subjects following weight loss.       

     Thiazolidinediones (TZDs), insulin, and other tyrosine phosphatase inhibitors are non-limiting examples of pharmaceutical agents that can be combined with or used in conjunction with the nucleic acid molecules (e.g. siRNA molecules) of the instant invention. Those skilled in the art will recognize that other drugs such as anti-diabetes and anti-obesity compounds and therapies can be similarly be readily combined with the nucleic acid molecules of the instant invention (e.g. siNA molecules) are hence within the scope of the instant invention. 
     Example 11 
     Multifunctional siNA Inhibition of PTP-1B RNA Expression 
     Multifunctional siNA Design 
     Once target sites have been identified for multifunctional siNA constructs, each strand of the siNA is designed with a complementary region of length, for example, of about 18 to about 28 nucleotides, that is complementary to a different target nucleic acid sequence. Each complementary region is designed with an adjacent flanking region of about 4 to about 22 nucleotides that is not complementary to the target sequence, but which comprises complementarity to the complementary region of the other sequence (see for example  FIG. 16 ). Hairpin constructs can likewise be designed (see for example  FIG. 17 ). Identification of complementary, palindrome or repeat sequences that are shared between the different target nucleic acid sequences can be used to shorten the overall length of the multifunctional siNA constructs (see for example  FIGS. 18 and 19 ). 
     In a non-limiting example, three additional categories of additional multifunctional siNA designs are presented that allow a single siNA molecule to silence multiple targets. The first method utilizes linkers to join siNAs (or multifunctional siNAs) in a direct manner. This can allow the most potent siNAs to be joined without creating a long, continuous stretch of RNA that has potential to trigger an interferon response. The second method is a dendrimeric extension of the overlapping or the linked multifunctional design; or alternatively the organization of siNA in a supramolecular format. The third method uses helix lengths greater than 30 base pairs. Processing of these siNAs by Dicer will reveal new, active 5′ antisense ends. Therefore, the long siNAs can target the sites defined by the original 5′ ends and those defined by the new ends that are created by Dicer processing. When used in combination with traditional multifunctional siNAs (where the sense and antisense strands each define a target) the approach can be used for example to target 4 or more sites. 
     I. Tethered Bifunctional siNAs 
     The basic idea is a novel approach to the design of multifunctional siNAs in which two antisense siNA strands are annealed to a single sense strand. The sense strand oligonucleotide contains a linker (e.g., non-nulcoetide linker as described herein) and two segments that anneal to the antisense siNA strands (see  FIG. 22 ). The linkers can also optionally comprise nucleotide-based linkers. Several potential advantages and variations to this approach include, but are not limited to:
     1. The two antisense siNAs are independent. Therefore, the choice of target sites is not constrained by a requirement for sequence conservation between two sites. Any two highly active siNAs can be combined to form a multifunctional siNA.   2. When used in combination with target sites having homology, siNAs that target a sequence present in two genes (e.g., different PTP-1B isoforms), the design can be used to target more than two sites. A single multifunctional siNA can be for example, used to target RNA of two different PTP-1B RNAs.   3. Multifunctional siNAs that use both the sense and antisense strands to target a gene can also be incorporated into a tethered multifuctional design. This leaves open the possibility of targeting 6 or more sites with a single complex.   4. It can be possible to anneal more than two antisense strand siNAs to a single tethered sense strand.   5. The design avoids long continuous stretches of dsRNA. Therefore, it is less likely to initiate an interferon response.   6. The linker (or modifications attached to it, such as conjugates described herein) can improve the pharmacokinetic properties of the complex or improve its incorporation into liposomes. Modifications introduced to the linker should not impact siNA activity to the same extent that they would if directly attached to the siNA (see for example  FIGS. 27 and 28 ).   7. The sense strand can extend beyond the annealed antisense strands to provide additional sites for the attachment of conjugates.   8. The polarity of the complex can be switched such that both of the antisense 3′ ends are adjacent to the linker and the 5′ ends are distal to the linker or combination thereof.
 
Dendrimer and Supramolecular siNAs
   

     In the dendrimer siNA approach, the synthesis of siNA is initiated by first synthesizing the dendrimer template followed by attaching various functional siNAs. Various constructs are depicted in  FIG. 23 . The number of functional siNAs that can be attached is only limited by the dimensions of the dendrimer used. 
     Supramolecular Approach to Multifunctional siNA 
     The supramolecular format simplifies the challenges of dendrimer synthesis. In this format, the siNA strands are synthesized by standard RNA chemistry, followed by annealing of various complementary strands. The individual strand synthesis contains an antisense sense sequence of one siNA at the 5′-end followed by a nucleic acid or synthetic linker, such as hexaethyleneglyol, which in turn is followed by sense strand of another siNA in 5′ to 3′ direction. Thus, the synthesis of siNA strands can be carried out in a standard 3′ to 5′ direction. Representative examples of trifunctional and tetrafunctional siNAs are depicted in  FIG. 24 . Based on a similar principle, higher functionality siNA constructs can be designed as long as efficient annealing of various strands is achieved. 
     Dicer Enabled Multifunctional siNA 
     Using bioinformatic analysis of multiple targets, stretches of identical sequences shared between differing target sequences can be identified ranging from about two to about fourteen nucleotides in length. These identical regions can be designed into extended siNA helixes (e.g., &gt;30 base pairs) such that the processing by Dicer reveals a secondary functional 5′-antisense site (see for example  FIG. 25 ). For example, when the first 17 nucleotides of a siNA antisense strand (e.g., 21 nucleotide strands in a duplex with 3′-TT overhangs) are complementary to a target RNA, robust silencing was observed at 25 nM. 80% silencing was observed with only 16 nucleotide complementarity in the same format. 
     Incorporation of this property into the designs of siNAs of about 30 to 40 or more base pairs results in additional multifunctional siNA constructs. The example in  FIG. 25  illustrates how a 30 base-pair duplex can target three distinct sequences after processing by Dicer-RNaseIII; these sequences can be on the same mRNA or separate RNAs, such as viral and host factor messages, or multiple points along a given pathway (e.g., inflammatory cascades). Furthermore, a 40 base-pair duplex can combine a bifunctional design in tandem, to provide a single duplex targeting four target sequences. An even more extensive approach can include use of homologous sequences to enable five or six targets silenced for one multifunctional duplex. The example in  FIG. 25  demonstrates how this can be achieved. A 30 base pair duplex is cleaved by Dicer into 22 and 8 base pair products from either end (8 b.p. fragments not shown). For ease of presentation the overhangs generated by dicer are not shown—but can be compensated for. Three targeting sequences are shown. The required sequence identity overlapped is indicated by grey boxes. The N&#39;s of the parent 30 b.p. siNA are suggested sites of 2′-OH positions to enable Dicer cleavage if this is tested in stabilized chemistries. Note that processing of a 30 mer duplex by Dicer RNase III does not give a precise 22+8 cleavage, but rather produces a series of closely related products (with 22+8 being the primary site). Therefore, processing by Dicer will yield a series of active siNAs. Another non-limiting example is shown in  FIG. 26 . A 40 base pair duplex is cleaved by Dicer into 20 base pair products from either end. For ease of presentation the overhangs generated by dicer are not shown—but can be compensated for. Four targeting sequences are shown in four colors, blue, light-blue and red and orange. The required sequence identity overlapped is indicated by grey boxes. This design format can be extended to larger RNAs. If chemically stabilized siNAs are bound by Dicer, then strategically located ribonucleotide linkages can enable designer cleavage products that permit our more extensive repertoire of multifunctional designs. For example cleavage products not limited to the Dicer standard of approximately 22-nucleotides can allow multifunctional siNA constructs with a target sequence identity overlap ranging from, for example, about 3 to about 15 nucleotides. 
     Example 12 
     Diagnostic Uses 
     The siNA molecules of the invention can be used in a variety of diagnostic applications, such as in the identification of molecular targets (e.g., RNA) in a variety of applications, for example, in clinical, industrial, environmental, agricultural and/or research settings. Such diagnostic use of siNA molecules involves utilizing reconstituted RNAi systems, for example, using cellular lysates or partially purified cellular lysates. siNA molecules of this invention can be used as diagnostic tools to examine genetic drift and mutations within diseased cells or to detect the presence of endogenous or exogenous, for example viral, RNA in a cell. The close relationship between siNA activity and the structure of the target RNA allows the detection of mutations in any region of the molecule, which alters the base-pairing and three-dimensional structure of the target RNA. By using multiple siNA molecules described in this invention, one can map nucleotide changes, which are important to RNA structure and function in vitro, as well as in cells and tissues. Cleavage of target RNAs with siNA molecules can be used to inhibit gene expression and define the role of specified gene products in the progression of disease or infection. In this manner, other genetic targets can be defined as important mediators of the disease. These experiments will lead to better treatment of the disease progression by affording the possibility of combination therapies (e.g., multiple siNA molecules targeted to different genes, siNA molecules coupled with known small molecule inhibitors, or intermittent treatment with combinations siNA molecules and/or other chemical or biological molecules). Other in vitro uses of siNA molecules of this invention are well known in the art, and include detection of the presence of mRNAs associated with a disease, infection, or related condition. Such RNA is detected by determining the presence of a cleavage product after treatment with a siNA using standard methodologies, for example, fluorescence resonance emission transfer (FRET). 
     In a specific example, siNA molecules that cleave only wild-type or mutant forms of the target RNA are used for the assay. The first siNA molecules (i.e., those that cleave only wild-type forms of target RNA) are used to identify wild-type RNA present in the sample and the second siNA molecules (i.e., those that cleave only mutant forms of target RNA) are used to identify mutant RNA in the sample. As reaction controls, synthetic substrates of both wild-type and mutant RNA are cleaved by both siNA molecules to demonstrate the relative siNA efficiencies in the reactions and the absence of cleavage of the “non-targeted” RNA species. The cleavage products from the synthetic substrates also serve to generate size markers for the analysis of wild-type and mutant RNAs in the sample population. Thus, each analysis requires two siNA molecules, two substrates and one unknown sample, which is combined into six reactions. The presence of cleavage products is determined using an RNase protection assay so that full-length and cleavage fragments of each RNA can be analyzed in one lane of a polyacrylamide gel. It is not absolutely required to quantify the results to gain insight into the expression of mutant RNAs and putative risk of the desired phenotypic changes in target cells. The expression of mRNA whose protein product is implicated in the development of the phenotype (i.e., disease related or infection related) is adequate to establish risk. If probes of comparable specific activity are used for both transcripts, then a qualitative comparison of RNA levels is adequate and decreases the cost of the initial diagnosis. Higher mutant form to wild-type ratios are correlated with higher risk whether RNA levels are compared qualitatively or quantitatively. 
     All patents and publications mentioned in the specification are indicative of the levels of skill of those skilled in the art to which the invention pertains. All references cited in this disclosure are incorporated by reference to the same extent as if each reference had been incorporated by reference in its entirety individually. 
     One skilled in the art would readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those inherent therein. The methods and compositions described herein as presently representative of preferred embodiments are exemplary and are not intended as limitations on the scope of the invention. Changes therein and other uses will occur to those skilled in the art, which are encompassed within the spirit of the invention, are defined by the scope of the claims. 
     It will be readily apparent to one skilled in the art that varying substitutions and modifications can be made to the invention disclosed herein without departing from the scope and spirit of the invention. Thus, such additional embodiments are within the scope of the present invention and the following claims. The present invention teaches one skilled in the art to test various combinations and/or substitutions of chemical modifications described herein toward generating nucleic acid constructs with improved activity for mediating RNAi activity. Such improved activity can comprise improved stability, improved bioavailability, and/or improved activation of cellular responses mediating RNAi. Therefore, the specific embodiments described herein are not limiting and one skilled in the art can readily appreciate that specific combinations of the modifications described herein can be tested without undue experimentation toward identifying siNA molecules with improved RNAi activity. 
     The invention illustratively described herein suitably can be practiced in the absence of any element or elements, limitation or limitations that are not specifically disclosed herein. Thus, for example, in each instance herein any of the terms “comprising”, “consisting essentially of”, and “consisting of” may be replaced with either of the other two terms. The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention that in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the present invention has been specifically disclosed by preferred embodiments, optional features, modification and variation of the concepts herein disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention as defined by the description and the appended claims. 
     In addition, where features or aspects of the invention are described in terms of Markush groups or other grouping of alternatives, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group or other group. 
     
       
         
           
               
             
               
                 TABLE I 
               
               
                   
               
               
                 PTP-1B Accession Numbers 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
            
               
                   
                 LOCUS PTPN1 3318 bp mRNA 
               
               
                   
                 linear PRI 09-JAN-2002 
               
               
                   
                 DEFINITION  Homo sapiens  protein tyrosine phosphatase, 
               
               
                   
                 non-receptor type 1 
               
               
                   
                 (PTPN1), mRNA. 
               
               
                   
                 ACCESSION NM_002827 
               
               
                   
                 LOCUS AY029236 2119 bp DNA 
               
               
                   
                 linear PRI 02-JUL-2001 
               
               
                   
                 DEFINITION  Homo sapiens  protein tyrosine phosphatase 
               
               
                   
                 1B (PTP-1B) gene, promoter 
               
               
                   
                 and partial cds. 
               
               
                   
                 ACCESSION AY029236 
               
               
                   
                 LOCUS AX418608 3247 bp DNA 
               
               
                   
                 linear PAT 18-JUN-2002 
               
               
                   
                 DEFINITION Sequence 3 from Patent WO0210378. 
               
               
                   
                 ACCESSION AX418608 
               
               
                   
                 VERSION AX418608.1 GI:21523469 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
               
             
               
                 TABLE II 
               
             
            
               
                   
               
               
                 PTP-1B siNA and Target Sequences 
                   
               
               
                 PPIB NM_000942 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                 Seq 
                   
                   
                 Seq 
                   
                   
                 Seq 
                   
               
               
                 Pos 
                 Seq 
                 ID 
                 UPos 
                 Upper seq 
                 ID 
                 LPos 
                 Lower seq 
                 ID 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                 1 
                 GUGAUGCGUAGUUCCGGCU 
                 1 
                 1 
                 GUGAUGCGUAGUUCCGGCU 
                 1 
                 19 
                 AGCCGGAACUACGCAUCAC 
                 186 
                   
               
               
                   
               
               
                 19 
                 UGCCGGUUGACAUGAAGAA 
                 2 
                 19 
                 UGCCGGUUGACAUGAAGAA 
                 2 
                 37 
                 UUCUUCAUGUCAACCGGCA 
                 187 
               
               
                   
               
               
                 37 
                 AGCAGCAGCGGCUAGGGCG 
                 3 
                 37 
                 AGCAGCAGCGGCUAGGGCG 
                 3 
                 55 
                 CGCCCUAGCCGCUGCUGCU 
                 188 
               
               
                   
               
               
                 55 
                 GGCGGUAGCUGCAGGGGUC 
                 4 
                 55 
                 GGCGGUAGCUGCAGGGGUC 
                 4 
                 73 
                 GACCCCUGCAGCUACCGCC 
                 189 
               
               
                   
               
               
                 73 
                 CGGGGAUUGCAGCGGGCCU 
                 5 
                 73 
                 CGGGGAUUGCAGCGGGCCU 
                 5 
                 91 
                 AGGCCCGCUGCAAUCCCCG 
                 190 
               
               
                   
               
               
                 91 
                 UCGGGGCUAAGAGCGCGAC 
                 6 
                 91 
                 UCGGGGCUAAGAGCGCGAC 
                 6 
                 109 
                 GUCGCGCUCUUAGCCCCGA 
                 191 
               
               
                   
               
               
                 109 
                 CGCGGCCUAGAGCGGCAGA 
                 7 
                 109 
                 CGCGGCCUAGAGCGGCAGA 
                 7 
                 127 
                 UCUGCCGCUCUAGGCCGCG 
                 192 
               
               
                   
               
               
                 127 
                 ACGGCGCAGUGGGCCGAGA 
                 8 
                 127 
                 ACGGCGCAGUGGGCCGAGA 
                 8 
                 145 
                 UCUCGGCCCACUGCGCCGU 
                 193 
               
               
                   
               
               
                 145 
                 AAGGAGGCGCAGCAGCCGC 
                 9 
                 145 
                 AAGGAGGCGCAGCAGCCGC 
                 9 
                 163 
                 GCGGCUGCUGCGCCUCCUU 
                 194 
               
               
                   
               
               
                 163 
                 CCCUGGCCCGUCAUGGAGA 
                 10 
                 163 
                 CCCUGGCCCGUCAUGGAGA 
                 10 
                 181 
                 UCUCCAUGACGGGCCAGGG 
                 195 
               
               
                   
               
               
                 181 
                 AUGGAAAAGGAGUUCGAGC 
                 11 
                 181 
                 AUGGAAAAGGAGUUCGAGC 
                 11 
                 199 
                 GCUCGAACUCCUUUUCCAU 
                 196 
               
               
                   
               
               
                 199 
                 CAGAUCGACAAGUCCGGGA 
                 12 
                 199 
                 CAGAUCGACAAGUCCGGGA 
                 12 
                 217 
                 UCCCGGACUUGUCGAUCUG 
                 197 
               
               
                   
               
               
                 217 
                 AGCUGGGCGGCCAUUUACC 
                 13 
                 217 
                 AGCUGGGCGGCCAUUUACC 
                 13 
                 235 
                 GGUAAAUGGCCGCCCAGCU 
                 198 
               
               
                   
               
               
                 235 
                 CAGGAUAUCCGACAUGAAG 
                 14 
                 235 
                 CAGGAUAUCCGACAUGAAG 
                 14 
                 253 
                 CUUCAUGUCGGAUAUCCUG 
                 199 
               
               
                   
               
               
                 253 
                 GCCAGUGACUUCCCAUGUA 
                 15 
                 253 
                 GCCAGUGACUUCCCAUGUA 
                 15 
                 271 
                 UACAUGGGAAGUCACUGGC 
                 200 
               
               
                   
               
               
                 271 
                 AGAGUGGCCAAGCUUCCUA 
                 16 
                 271 
                 AGAGUGGCCAAGCUUCCUA 
                 16 
                 289 
                 UAGGAAGCUUGGCCACUCU 
                 201 
               
               
                   
               
               
                 289 
                 AAGAACAAAAACCGAAAUA 
                 17 
                 289 
                 AAGAACAAAAACCGAAAUA 
                 17 
                 307 
                 UAUUUCGGUUUUUGUUCUU 
                 202 
               
               
                   
               
               
                 307 
                 AGGUACAGAGACGUCAGUC 
                 18 
                 307 
                 AGGUACAGAGACGUCAGUC 
                 18 
                 325 
                 GACUGACGUCUCUGUACCU 
                 203 
               
               
                   
               
               
                 325 
                 CCCUUUGACCAUAGUCGGA 
                 19 
                 325 
                 CCCUUUGACCAUAGUCGGA 
                 19 
                 343 
                 UCCGACUAUGGUCAAAGGG 
                 204 
               
               
                   
               
               
                 343 
                 AUUAAACUACAUCAAGAAG 
                 20 
                 343 
                 AUUAAACUACAUCAAGAAG 
                 20 
                 361 
                 CUUCUUGAUGUAGUUUAAU 
                 205 
               
               
                   
               
               
                 361 
                 GAUAAUGACUAUAUCAACG 
                 21 
                 361 
                 GAUAAUGACUAUAUCAACG 
                 21 
                 379 
                 CGUUGAUAUAGUCAUUAUC 
                 206 
               
               
                   
               
               
                 379 
                 GCUAGUUUGAUAAAAAUGG 
                 22 
                 379 
                 GCUAGUUUGAUAAAAAUGG 
                 22 
                 397 
                 CCAUUUUUAUCAAACUAGC 
                 207 
               
               
                   
               
               
                 397 
                 GAAGAAGCCCAAAGGAGUU 
                 23 
                 397 
                 GAAGAAGCCCAAAGGAGUU 
                 23 
                 415 
                 AACUCCUUUGGGCUUCUUC 
                 208 
               
               
                   
               
               
                 415 
                 UACAUUCUUACCCAGGGCC 
                 24 
                 415 
                 UACAUUCUUACCCAGGGCC 
                 24 
                 433 
                 GGCCCUGGGUAAGAAUGUA 
                 209 
               
               
                   
               
               
                 433 
                 CCUUUGCCUAACACAUGCG 
                 25 
                 433 
                 CCUUUGCCUAACACAUGCG 
                 25 
                 451 
                 CGCAUGUGUUAGGCAAAGG 
                 210 
               
               
                   
               
               
                 451 
                 GGUCACUUUUGGGAGAUGG 
                 26 
                 451 
                 GGUCACUUUUGGGAGAUGG 
                 26 
                 469 
                 CCAUCUCCCAAAAGUGACC 
                 211 
               
               
                   
               
               
                 469 
                 GUGUGGGAGCAGAAAAGCA 
                 27 
                 469 
                 GUGUGGGAGCAGAAAAGCA 
                 27 
                 487 
                 UGCUUUUCUGCUCCCACAC 
                 212 
               
               
                   
               
               
                 487 
                 AGGGGUGUCGUCAUGCUCA 
                 28 
                 487 
                 AGGGGUGUCGUCAUGCUCA 
                 28 
                 505 
                 UGAGCAUGACGACACCCCU 
                 213 
               
               
                   
               
               
                 505 
                 AACAGAGUGAUGGAGAAAG 
                 29 
                 505 
                 AACAGAGUGAUGGAGAAAG 
                 29 
                 523 
                 CUUUCUCCAUCACUCUGUU 
                 214 
               
               
                   
               
               
                 523 
                 GGUUCGUUAAAAUGCGCAC 
                 30 
                 523 
                 GGUUCGUUAAAAUGCGCAC 
                 30 
                 541 
                 GUGCGCAUUUUAACGAACC 
                 215 
               
               
                   
               
               
                 541 
                 CAAUACUGGCCACAAAAAG 
                 31 
                 541 
                 CAAUACUGGCCACAAAAAG 
                 31 
                 559 
                 CUUUUUGUGGCCAGUAUUG 
                 216 
               
               
                   
               
               
                 559 
                 GAAGAAAAAGAGAUGAUCU 
                 32 
                 559 
                 GAAGAAAAAGAGAUGAUCU 
                 32 
                 577 
                 AGAUCAUCUCUUUUUCUUC 
                 217 
               
               
                   
               
               
                 577 
                 UUUGAAGACACAAAUUUGA 
                 33 
                 577 
                 UUUGAAGACACAAAUUUGA 
                 33 
                 595 
                 UCAAAUUUGUGUCUUCAAA 
                 218 
               
               
                   
               
               
                 595 
                 AAAUUAACAUUGAUCUCUG 
                 34 
                 595 
                 AAAUUAACAUUGAUCUCUG 
                 34 
                 613 
                 CAGAGAUCAAUGUUAAUUU 
                 219 
               
               
                   
               
               
                 613 
                 GAAGAUAUCAAGUCAUAUU 
                 35 
                 613 
                 GAAGAUAUCAAGUCAUAUU 
                 35 
                 631 
                 AAUAUGACUUGAUAUCUUC 
                 220 
               
               
                   
               
               
                 631 
                 UAUACAGUGCGACAGCUAG 
                 36 
                 631 
                 UAUACAGUGCGACAGCUAG 
                 36 
                 649 
                 CUAGCUGUCGCACUGUAUA 
                 221 
               
               
                   
               
               
                 649 
                 GAAUUGGAAAACCUUACAA 
                 37 
                 649 
                 GAAUUGGAAAACCUUACAA 
                 37 
                 667 
                 UUGUAAGGUUUUCCAAUUC 
                 222 
               
               
                   
               
               
                 667 
                 ACCCAAGAAACUCGAGAGA 
                 38 
                 667 
                 ACCCAAGAAACUCGAGAGA 
                 38 
                 685 
                 UCUCUCGAGUUUCUUGGGU 
                 223 
               
               
                   
               
               
                 685 
                 AUCUUACAUUUCCACUAUA 
                 39 
                 685 
                 AUCUUACAUUUCCACUAUA 
                 39 
                 703 
                 UAUAGUGGAAAUGUAAGAU 
                 224 
               
               
                   
               
               
                 703 
                 ACCACAUGGCCUGACUUUG 
                 40 
                 703 
                 ACCACAUGGCCUGACUUUG 
                 40 
                 721 
                 CAAAGUCAGGCCAUGUGGU 
                 225 
               
               
                   
               
               
                 721 
                 GGAGUCCCUGAAUCACCAG 
                 41 
                 721 
                 GGAGUCCCUGAAUCACCAG 
                 41 
                 739 
                 CUGGUGAUUCAGGGACUCC 
                 226 
               
               
                   
               
               
                 739 
                 GCCUCAUUCUUGAACUUUC 
                 42 
                 739 
                 GCCUCAUUCUUGAACUUUC 
                 42 
                 757 
                 GAAAGUUCAAGAAUGAGGC 
                 227 
               
               
                   
               
               
                 757 
                 CUUUUCAAAGUCCGAGAGU 
                 43 
                 757 
                 CUUUUCAAAGUCCGAGAGU 
                 43 
                 775 
                 ACUCUCGGACUUUGAAAAG 
                 228 
               
               
                   
               
               
                 775 
                 UCAGGGUCACUCAGCCCGG 
                 44 
                 775 
                 UCAGGGUCACUCAGCCCGG 
                 44 
                 793 
                 CCGGGCUGAGUGACCCUGA 
                 229 
               
               
                   
               
               
                 793 
                 GAGCACGGGCCCGUUGUGG 
                 45 
                 793 
                 GAGCACGGGCCCGUUGUGG 
                 45 
                 811 
                 CCACAACGGGCCCGUGCUC 
                 230 
               
               
                   
               
               
                 811 
                 GUGCACUGCAGUGCAGGCA 
                 46 
                 811 
                 GUGCACUGCAGUGCAGGCA 
                 46 
                 829 
                 UGCCUGCACUGCAGUGCAC 
                 231 
               
               
                   
               
               
                 829 
                 AUCGGCAGGUCUGGAACCU 
                 47 
                 829 
                 AUCGGCAGGUCUGGAACCU 
                 47 
                 847 
                 AGGUUCCAGACCUGCCGAU 
                 232 
               
               
                   
               
               
                 847 
                 UUCUGUCUGGCUGAUACCU 
                 48 
                 847 
                 UUCUGUCUGGCUGAUACCU 
                 48 
                 865 
                 AGGUAUCAGCCAGACAGAA 
                 233 
               
               
                   
               
               
                 865 
                 UGCCUCUUGCUGAUGGACA 
                 49 
                 865 
                 UGCCUCUUGCUGAUGGACA 
                 49 
                 883 
                 UGUCCAUCAGCAAGAGGCA 
                 234 
               
               
                   
               
               
                 883 
                 AAGAGGAAAGACCCUUCUU 
                 50 
                 883 
                 AAGAGGAAAGACCCUUCUU 
                 50 
                 901 
                 AAGAAGGGUCUUUCCUCUU 
                 235 
               
               
                   
               
               
                 901 
                 UCCGUUGAUAUCAAGAAAG 
                 51 
                 901 
                 UCCGUUGAUAUCAAGAAAG 
                 51 
                 919 
                 CUUUCUUGAUAUCAACGGA 
                 236 
               
               
                   
               
               
                 919 
                 GUGCUGUUAGAAAUGAGGA 
                 52 
                 919 
                 GUGCUGUUAGAAAUGAGGA 
                 52 
                 937 
                 UCCUCAUUUCUAACAGCAC 
                 237 
               
               
                   
               
               
                 937 
                 AAGUUUCGGAUGGGGCUGA 
                 53 
                 937 
                 AAGUUUCGGAUGGGGCUGA 
                 53 
                 955 
                 UCAGCCCCAUCCGAAACUU 
                 238 
               
               
                   
               
               
                 955 
                 AUCCAGACAGCCGACCAGC 
                 54 
                 955 
                 AUCCAGACAGCCGACCAGC 
                 54 
                 973 
                 GCUGGUCGGCUGUCUGGAU 
                 239 
               
               
                   
               
               
                 973 
                 CUGCGCUUCUCCUACCUGG 
                 55 
                 973 
                 CUGCGCUUCUCCUACCUGG 
                 55 
                 991 
                 CCAGGUAGGAGAAGCGCAG 
                 240 
               
               
                   
               
               
                 991 
                 GCUGUGAUCGAAGGUGCCA 
                 56 
                 991 
                 GCUGUGAUCGAAGGUGCCA 
                 56 
                 1009 
                 UGGCACCUUCGAUCACAGC 
                 241 
               
               
                   
               
               
                 1009 
                 AAAUUCAUCAUGGGGGACU 
                 57 
                 1009 
                 AAAUUCAUCAUGGGGGACU 
                 57 
                 1027 
                 AGUCCCCCAUGAUGAAUUU 
                 242 
               
               
                   
               
               
                 1027 
                 UCUUCCGUGCAGGAUCAGU 
                 58 
                 1027 
                 UCUUCCGUGCAGGAUCAGU 
                 58 
                 1045 
                 ACUGAUCCUGCACGGAAGA 
                 243 
               
               
                   
               
               
                 1045 
                 UGGAAGGAGCUUUCCCACG 
                 59 
                 1045 
                 UGGAAGGAGCUUUCCCACG 
                 59 
                 1063 
                 CGUGGGAAAGCUCCUUCCA 
                 244 
               
               
                   
               
               
                 1063 
                 GAGGACCUGGAGCCCCCAC 
                 60 
                 1063 
                 GAGGACCUGGAGCCCCCAC 
                 60 
                 1081 
                 GUGGGGGCUCCAGGUCCUC 
                 245 
               
               
                   
               
               
                 1081 
                 CCCGAGCAUAUCCCCCCAC 
                 61 
                 1081 
                 CCCGAGCAUAUCCCCCCAC 
                 61 
                 1099 
                 GUGGGGGGAUAUGCUCGGG 
                 246 
               
               
                   
               
               
                 1099 
                 CCUCCCCGGCCACCCAAAC 
                 62 
                 1099 
                 CCUCCCCGGCCACCCAAAC 
                 62 
                 1117 
                 GUUUGGGUGGCCGGGGAGG 
                 247 
               
               
                   
               
               
                 1117 
                 CGAAUCCUGGAGCCACACA 
                 63 
                 1117 
                 CGAAUCCUGGAGCCACACA 
                 63 
                 1135 
                 UGUGUGGCUCCAGGAUUCG 
                 248 
               
               
                   
               
               
                 1135 
                 AAUGGGAAAUGCAGGGAGU 
                 64 
                 1135 
                 AAUGGGAAAUGCAGGGAGU 
                 64 
                 1153 
                 ACUCCCUGCAUUUCCCAUU 
                 249 
               
               
                   
               
               
                 1153 
                 UUCUUCCCAAAUCACCAGU 
                 65 
                 1153 
                 UUCUUCCCAAAUCACCAGU 
                 65 
                 1171 
                 ACUGGUGAUUUGGGAAGAA 
                 250 
               
               
                   
               
               
                 1171 
                 UGGGUGAAGGAAGAGACCC 
                 66 
                 1171 
                 UGGGUGAAGGAAGAGACCC 
                 66 
                 1189 
                 GGGUCUCUUCCUUCACCCA 
                 251 
               
               
                   
               
               
                 1189 
                 CAGGAGGAUAAAGACUGCC 
                 67 
                 1189 
                 CAGGAGGAUAAAGACUGCC 
                 67 
                 1207 
                 GGCAGUGUUUAUCCUCCUG 
                 252 
               
               
                   
               
               
                 1207 
                 CCCAUCAAGGAAGAAAAAG 
                 68 
                 1207 
                 CCCAUCAAGGAAGAAAAAG 
                 68 
                 1225 
                 CUUUUUCUUCCUUGAUGGG 
                 253 
               
               
                   
               
               
                 1225 
                 GGAAGCCCCUUAAAUGCCG 
                 69 
                 1225 
                 GGAAGCCCCUUAAAUGCCG 
                 69 
                 1243 
                 CGGCAUUUAAGGGGCUUCC 
                 254 
               
               
                   
               
               
                 1243 
                 GCACCCUACGGCAUCGAAA 
                 70 
                 1243 
                 GCACCCUACGGCAUCGAAA 
                 70 
                 1261 
                 UUUCGAUGCCGUAGGGUGC 
                 255 
               
               
                   
               
               
                 1261 
                 AGCAUGAGUCAAGACACUG 
                 71 
                 1261 
                 AGCAUGAGUCAAGACACUG 
                 71 
                 1279 
                 CAGUGUCUUGACUCAUGCU 
                 256 
               
               
                   
               
               
                 1279 
                 GAAGUUAGAAGUCGGGUCG 
                 72 
                 1279 
                 GAAGUUAGAAGUCGGGUCG 
                 72 
                 1297 
                 CGACCCGACUUCUAACUUC 
                 257 
               
               
                   
               
               
                 1297 
                 GUGGGGGGAAGUCUUCGAG 
                 73 
                 1297 
                 GUGGGGGGAAGUCUUCGAG 
                 73 
                 1315 
                 CUCGAAGACUUCCCCCCAC 
                 258 
               
               
                   
               
               
                 1315 
                 GGUGCCCAGGCUGCCUCCC 
                 74 
                 1315 
                 GGUGCCCAGGCUGCCUCCC 
                 74 
                 1333 
                 GGGAGGCAGCCUGGGCACC 
                 259 
               
               
                   
               
               
                 1333 
                 CCAGCCAAAGGGGAGCCGU 
                 75 
                 1333 
                 CCAGCCAAAGGGGAGCCGU 
                 75 
                 1351 
                 ACGGCUCCCCUUUGGCUGG 
                 260 
               
               
                   
               
               
                 1351 
                 UCACUGCCCGAGAAGGACG 
                 76 
                 1351 
                 UCACUGCCCGAGAAGGACG 
                 76 
                 1369 
                 CGUCCUUCUCGGGCAGUGA 
                 261 
               
               
                   
               
               
                 1369 
                 GAGGACCAUGCACUGAGUU 
                 77 
                 1369 
                 GAGGACCAUGCACUGAGUU 
                 77 
                 1387 
                 AACUCAGUGCAUGGUCCUC 
                 262 
               
               
                   
               
               
                 1387 
                 UACUGGAAGCCCUUCCUGG 
                 78 
                 1387 
                 UACUGGAAGCCCUUCCUGG 
                 78 
                 1405 
                 CCAGGAAGGGCUUCCAGUA 
                 263 
               
               
                   
               
               
                 1405 
                 GUCAACAUGUGCGUGGCUA 
                 79 
                 1405 
                 GUCAACAUGUGCGUGGCUA 
                 79 
                 1423 
                 UAGCCACGCACAUGUUGAC 
                 264 
               
               
                   
               
               
                 1423 
                 ACGGUCCUCACGGCCGGCG 
                 80 
                 1423 
                 ACGGUCCUCACGGCCGGCG 
                 80 
                 1441 
                 CGCCGGCCGUGAGGACCGU 
                 265 
               
               
                   
               
               
                 1441 
                 GCUUACCUCUGCUACAGGU 
                 81 
                 1441 
                 GCUUACCUCUGCUACAGGU 
                 81 
                 1459 
                 ACCUGUAGCAGAGGUAAGC 
                 266 
               
               
                   
               
               
                 1459 
                 UUCCUGUUCAACAGCAACA 
                 82 
                 1459 
                 UUCCUGUUCAACAGCAACA 
                 82 
                 1477 
                 UGUUGCUGUUGAACAGGAA 
                 267 
               
               
                   
               
               
                 1477 
                 ACAUAGCCUGACCCUCCUC 
                 83 
                 1477 
                 ACAUAGCCUGACCCUCCUC 
                 83 
                 1495 
                 GAGGAGGGUCAGGCUAUGU 
                 268 
               
               
                   
               
               
                 1495 
                 CCACUCCACCUCCACCCAC 
                 84 
                 1495 
                 CCACUCCACCUCCACCCAC 
                 84 
                 1513 
                 GUGGGUGGAGGUGGAGUGG 
                 269 
               
               
                   
               
               
                 1513 
                 CUGUCCGCCUCUGGCCGCA 
                 85 
                 1513 
                 GUGUCCGCCUCUGCCCGCA 
                 85 
                 1531 
                 UGCGGGCAGAGGCGGACAG 
                 270 
               
               
                   
               
               
                 1531 
                 AGAGCCCACGCCCGACUAG 
                 86 
                 1531 
                 AGAGCCCACGCCCGACUAG 
                 86 
                 1549 
                 CUAGUCGGGCGUGGGCUCU 
                 271 
               
               
                   
               
               
                 1549 
                 GCAGGCAUGCCGCGGUAGG 
                 87 
                 1549 
                 GCAGGCAUGCCGCGGUAGG 
                 87 
                 1567 
                 CCUACCGCGGCAUGCCUGC 
                 272 
               
               
                   
               
               
                 1567 
                 GUAAGGGCCGCCGGACCGC 
                 88 
                 1567 
                 GUAAGGGCCGCCGGACCGC 
                 88 
                 1585 
                 GCGGUCCGGCGGCCCUUAC 
                 273 
               
               
                   
               
               
                 1585 
                 CGUAGAGAGCCGGGCCCCG 
                 89 
                 1585 
                 CGUAGAGAGCCGGGCCCCG 
                 89 
                 1603 
                 CGGGGCCCGGCUCUCUACG 
                 274 
               
               
                   
               
               
                 1603 
                 GGACGGACGUUGGUUCUGC 
                 90 
                 1603 
                 GGACGGACGUUGGUUCUGC 
                 90 
                 1621 
                 GCAGAACCAACGUCCGUCC 
                 275 
               
               
                   
               
               
                 1621 
                 CACUAAAACCCAUCUUCCC 
                 91 
                 1621 
                 CACUAAAACCCAUCUUCCC 
                 91 
                 1639 
                 GGGAAGAUGGGUUUUAGUG 
                 276 
               
               
                   
               
               
                 1639 
                 CCGGAUGUGUGUCUCACCC 
                 92 
                 1639 
                 CCGGAUGUGUGUCUCACCC 
                 92 
                 1657 
                 GGGUGAGACACACAUCCGG 
                 277 
               
               
                   
               
               
                 1657 
                 CCUCAUCCUUUUACUUUUU 
                 93 
                 1657 
                 CCUCAUCCUUUUACUUUUU 
                 93 
                 1675 
                 AAAAAGUAAAAGGAUGAGG 
                 278 
               
               
                   
               
               
                 1675 
                 UGCCCCUUCCACUUUGAGU 
                 94 
                 1675 
                 UGCCCCUUCCACUUUGAGU 
                 94 
                 1693 
                 ACUCAAAGUGGAAGGGGCA 
                 279 
               
               
                   
               
               
                 1693 
                 UACCAAAUCCACAAGCCAU 
                 95 
                 1693 
                 UACCAAAUCCACAAGCCAU 
                 95 
                 1711 
                 AUGGCUUGUGGAUUUGGUA 
                 280 
               
               
                   
               
               
                 1711 
                 UUUUUUGAGGAGAGUGAAA 
                 96 
                 1711 
                 UUUUUUGAGGAGAGUGAAA 
                 96 
                 1729 
                 UUUCACUCUCCUCAAAAAA 
                 281 
               
               
                   
               
               
                 1729 
                 AGAGAGUACCAUGGUGGCG 
                 97 
                 1729 
                 AGAGAGUACCAUGCUGGCG 
                 97 
                 1747 
                 CGCCAGCAUGGUACUCUCU 
                 282 
               
               
                   
               
               
                 1747 
                 GGCGCAGAGGGAAGGGGCC 
                 98 
                 1747 
                 GGCGCAGAGGGAAGGGGCC 
                 98 
                 1765 
                 GGCCCCUUCCCUCUGCGCC 
                 283 
               
               
                   
               
               
                 1765 
                 CUACACCCGUCUUGGGGCU 
                 99 
                 1765 
                 CUACACCCGUCUUGGGGCU 
                 99 
                 1783 
                 AGCCCCAAGACGGGUGUAG 
                 284 
               
               
                   
               
               
                 1783 
                 UCGCCCCACCCAGGGCUCC 
                 100 
                 1783 
                 UCGCCCCACCCAGGGCUCC 
                 100 
                 1801 
                 GGAGCCCUGGGUGGGGCGA 
                 285 
               
               
                   
               
               
                 1801 
                 CCUCCUGGAGCAUCCCAGG 
                 101 
                 1801 
                 CCUCCUGGAGCAUCCCAGG 
                 101 
                 1819 
                 CCUGGGAUGCUCCAGGAGG 
                 286 
               
               
                   
               
               
                 1819 
                 GCGGGCGGCACGCCAACAG 
                 102 
                 1819 
                 GCGGGCGGCACGCCAACAG 
                 102 
                 1837 
                 CUGUUGGCGUGCCGCCCGC 
                 287 
               
               
                   
               
               
                 1837 
                 GCCCCCCCCUUGAAUCUGC 
                 103 
                 1837 
                 GCCCCCCCCUUGAAUCUGC 
                 103 
                 1855 
                 GCAGAUUCAAGGGGGGGGC 
                 288 
               
               
                   
               
               
                 1855 
                 CAGGGAGCAACUCUCCACU 
                 104 
                 1855 
                 CAGGGAGCAACUCUCCACU 
                 104 
                 1873 
                 AGUGGAGAGUUGCUCCCUG 
                 289 
               
               
                   
               
               
                 1873 
                 UCCAUAUUUAUUUAAACAA 
                 105 
                 1873 
                 UCCAUAUUUAUUUAAACAA 
                 105 
                 1891 
                 UUGUUUAAAUAAAUAUGGA 
                 290 
               
               
                   
               
               
                 1891 
                 AUUUUUUCCCCAAAGGCAU 
                 106 
                 1891 
                 AUUUUUUCCCCAAAGGCAU 
                 106 
                 1909 
                 AUGCCUUUGGGGAAAAAAU 
                 291 
               
               
                   
               
               
                 1909 
                 UCCAUAGUGCACUAGCAUU 
                 107 
                 1909 
                 UCCAUAGUGCACUAGCAUU 
                 107 
                 1927 
                 AAUGCUAGUGCACUAUGGA 
                 292 
               
               
                   
               
               
                 1927 
                 UUUCUUGAACCAAUAAUGU 
                 108 
                 1927 
                 UUUCUUGAACCAAUAAUGU 
                 108 
                 1945 
                 ACAUUAUUGGUUCAAGAAA 
                 293 
               
               
                   
               
               
                 1945 
                 UAUUAAAAUUUUUUGAUGU 
                 109 
                 1945 
                 UAUUAAAAUUUUUUGAUGU 
                 109 
                 1963 
                 ACAUCAAAAAAUUUUAAUA 
                 294 
               
               
                   
               
               
                 1963 
                 UCAGCCUUGCAUCAAGGGC 
                 110 
                 1963 
                 UCAGCCUUGCAUCAAGGGC 
                 110 
                 1981 
                 GCCCUUGAUGCAAGGCUGA 
                 295 
               
               
                   
               
               
                 1981 
                 CUUUAUCAAAAAGUACAAU 
                 111 
                 1981 
                 CUUUAUCAAAAAGUACAAU 
                 111 
                 1999 
                 AUUGUACUUUUUGAUAAAG 
                 296 
               
               
                   
               
               
                 1999 
                 UAAUAAAUCCUCAGGUAGU 
                 112 
                 1999 
                 UAAUAAAUCCUCAGGUAGU 
                 112 
                 2017 
                 ACUACCUGAGGAUUUAUUA 
                 297 
               
               
                   
               
               
                 2017 
                 UACUGGGAAUGGAAGGCUU 
                 113 
                 2017 
                 UACUGGGAAUGGAAGGCUU 
                 113 
                 2035 
                 AAGCCUUCCAUUCCCAGUA 
                 298 
               
               
                   
               
               
                 2035 
                 UUGCCAUGGGCCUGCUGCG 
                 114 
                 2035 
                 UUGCCAUGGGCCUGCUGCG 
                 114 
                 2053 
                 CGCAGCAGGCCCAUGGCAA 
                 299 
               
               
                   
               
               
                 2053 
                 GUCAGACCAGUACUGGGAA 
                 115 
                 2053 
                 GUCAGACCAGUACUGGGAA 
                 115 
                 2071 
                 UUCCCAGUACUGGUCUGAC 
                 300 
               
               
                   
               
               
                 2071 
                 AGGAGGACGGUUGUAAGCA 
                 116 
                 2071 
                 AGGAGGACGGUUGUAAGCA 
                 116 
                 2089 
                 UGCUUACAACCGUCCUCCU 
                 301 
               
               
                   
               
               
                 2089 
                 AGUUGUUAUUUAGUGAUAU 
                 117 
                 2089 
                 AGUUGUUAUUUAGUGAUAU 
                 117 
                 2107 
                 AUAUCACUAAAUAACAACU 
                 302 
               
               
                   
               
               
                 2107 
                 UUGUGGGUAACGUGAGAAG 
                 118 
                 2107 
                 UUGUGGGUAACGUGAGAAG 
                 118 
                 2125 
                 CUUCUCACGUUACCCACAA 
                 303 
               
               
                   
               
               
                 2125 
                 GAUAGAACAAUGCUAUAAU 
                 119 
                 2125 
                 GAUAGAACAAUGCUAUAAU 
                 119 
                 2143 
                 AUUAUAGCAUUGUUCUAUC 
                 304 
               
               
                   
               
               
                 2143 
                 UAUAUAAUGAACACGUGGG 
                 120 
                 2143 
                 UAUAUAAUGAACACGUGGG 
                 120 
                 2161 
                 CCCACGUGUUCAUUAUAUA 
                 305 
               
               
                   
               
               
                 2161 
                 GUAUUUAAUAAGAAACAUG 
                 121 
                 2161 
                 GUAUUUAAUAAGAAACAUG 
                 121 
                 2179 
                 CAUGUUUCUUAUUAAAUAC 
                 306 
               
               
                   
               
               
                 2179 
                 GAUGUGAGAUUACUUUGUC 
                 122 
                 2179 
                 GAUGUGAGAUUACUUUGUC 
                 122 
                 2197 
                 GACAAAGUAAUCUCACAUC 
                 307 
               
               
                   
               
               
                 2197 
                 CCCGCUUAUUCUCCUCCCU 
                 123 
                 2197 
                 CCCGCUUAUUCUCCUCCCU 
                 123 
                 2215 
                 AGGGAGGAGAAUAAGCGGG 
                 308 
               
               
                   
               
               
                 2215 
                 UGUUAUCUGCUAGAUCUAG 
                 124 
                 2215 
                 UGUUAUCUGCUAGAUCUAG 
                 124 
                 2233 
                 CUAGAUCUAGCAGAUAACA 
                 309 
               
               
                   
               
               
                 2233 
                 GUUCUCAAUCACUGCUCCC 
                 125 
                 2233 
                 GUUCUCAAUCACUGCUCCC 
                 125 
                 2251 
                 GGGAGCAGUGAUUGAGAAC 
                 310 
               
               
                   
               
               
                 2251 
                 CCCGUGUGUAUUAGAAUGC 
                 126 
                 2251 
                 CCCGUGUGUAUUAGAAUGC 
                 126 
                 2269 
                 GCAUUCUAAUACACACGGG 
                 311 
               
               
                   
               
               
                 2269 
                 CAUGUAAGGUCUUCUUGUG 
                 127 
                 2269 
                 CAUGUAAGGUCUUCUUGUG 
                 127 
                 2287 
                 CACAAGAAGACCUUACAUG 
                 312 
               
               
                   
               
               
                 2287 
                 GUCCUGAUGAAAAAUAUGU 
                 128 
                 2287 
                 GUCCUGAUGAAAAAUAUGU 
                 128 
                 2305 
                 ACAUAUUUUUCAUCAGGAC 
                 313 
               
               
                   
               
               
                 2305 
                 UGCUUGAAAUGAGAAACUU 
                 129 
                 2305 
                 UGCUUGAAAUGAGAAACUU 
                 129 
                 2323 
                 AAGUUUCUCAUUUCAAGCA 
                 314 
               
               
                   
               
               
                 2323 
                 UUGAUCUCUGCUUACUAAU 
                 130 
                 2323 
                 UUGAUCUCUGCUUACUAAU 
                 130 
                 2341 
                 AUUAGUAAGCAGAGAUCAA 
                 315 
               
               
                   
               
               
                 2341 
                 UGUGCCCCAUGUCCAAGUC 
                 131 
                 2341 
                 UGUGCCCCAUGUCCAAGUC 
                 131 
                 2359 
                 GACUUGGACAUGGGGCACA 
                 316 
               
               
                   
               
               
                 2359 
                 CCAACCUGCCUGUGCAUGA 
                 132 
                 2359 
                 CCAACCUGCCUGUGCAUGA 
                 132 
                 2377 
                 UCAUGCACAGGCAGGUUGG 
                 317 
               
               
                   
               
               
                 2377 
                 ACCUGAUCAUUACAUGGCU 
                 133 
                 2377 
                 ACCUGAUCAUUACAUGGCU 
                 133 
                 2395 
                 AGCCAUGUAAUGAUCAGGU 
                 318 
               
               
                   
               
               
                 2395 
                 UGUGGUUCCUAAGCCUGUU 
                 134 
                 2395 
                 UGUGGUUCCUAAGCCUGUU 
                 134 
                 2413 
                 AACAGGCUUAGGAACCACA 
                 319 
               
               
                   
               
               
                 2413 
                 UGCUGAAGUCAUUGUCGCU 
                 135 
                 2413 
                 UGCUGAAGUCAUUGUCGCU 
                 135 
                 2431 
                 AGCGACAAUGACUUCAGCA 
                 320 
               
               
                   
               
               
                 2431 
                 UCAGCAAUAGGGUGCAGUU 
                 136 
                 2431 
                 UCAGCAAUAGGGUGCAGUU 
                 136 
                 2449 
                 AACUGCACCCUAUUGCUGA 
                 321 
               
               
                   
               
               
                 2449 
                 UUUCCAGGAAUAGGCAUUU 
                 137 
                 2449 
                 UUUCCAGGAAUAGGCAUUU 
                 137 
                 2467 
                 AAAUGCCUAUUCCUGGAAA 
                 322 
               
               
                   
               
               
                 2467 
                 UGCCUAAUUCCUGGCAUGA 
                 138 
                 2467 
                 UGCCUAAUUCCUGGCAUGA 
                 138 
                 2485 
                 UCAUGCCAGGAAUUAGGCA 
                 323 
               
               
                   
               
               
                 2485 
                 ACACUCUAGUGACUUCCUG 
                 139 
                 2485 
                 ACACUCUAGUGACUUCCUG 
                 139 
                 2503 
                 CAGGAAGUCACUAGAGUGU 
                 324 
               
               
                   
               
               
                 2503 
                 GGUGAGGCCCAGCCUGUCC 
                 140 
                 2503 
                 GGUGAGGCCCAGCCUGUCC 
                 140 
                 2521 
                 GGACAGGCUGGGCCUCACC 
                 325 
               
               
                   
               
               
                 2521 
                 CUGGUACAGCAGGGUCUUG 
                 141 
                 2521 
                 CUGGUACAGCAGGGUCUUG 
                 141 
                 2539 
                 CAAGACCCUGCUGUACCAG 
                 326 
               
               
                   
               
               
                 2539 
                 GCUGUAACUCAGACAUUCC 
                 142 
                 2539 
                 GCUGUAACUCAGACAUUCC 
                 142 
                 2557 
                 GGAAUGUCUGAGUUACAGC 
                 327 
               
               
                   
               
               
                 2557 
                 CAAGGGUAUGGGAAGCCAU 
                 143 
                 2557 
                 CAAGGGUAUGGGAAGCCAU 
                 143 
                 2575 
                 AUGGCUUCCCAUACCCUUG 
                 328 
               
               
                   
               
               
                 2575 
                 UAUUCACACCUCACGCUCU 
                 144 
                 2575 
                 UAUUCACACCUCACGCUCU 
                 144 
                 2593 
                 AGAGCGUGAGGUGUGAAUA 
                 329 
               
               
                   
               
               
                 2593 
                 UGGACAUGAUUUAGGGAAG 
                 145 
                 2593 
                 UGGACAUGAUUUAGGGAAG 
                 145 
                 2611 
                 CUUCCCUAAAUCAUGUCCA 
                 330 
               
               
                   
               
               
                 2611 
                 GCAGGGACACCCCCCGCCC 
                 146 
                 2611 
                 GCAGGGACACCCCCCGCCC 
                 146 
                 2629 
                 GGGCGGGGGGUGUCCCUGC 
                 331 
               
               
                   
               
               
                 2629 
                 CCCCACCUUUGGGAUCAGC 
                 147 
                 2629 
                 CCCCACCUUUGGGAUCAGC 
                 147 
                 2647 
                 GCUGAUCCCAAAGGUGGGG 
                 332 
               
               
                   
               
               
                 2647 
                 CCUCCGCCAUUCCAAGUCA 
                 148 
                 2647 
                 CCUCCGCCAUUCCAAGUCA 
                 148 
                 2665 
                 UGACUUGGAAUGGCGGAGG 
                 333 
               
               
                   
               
               
                 2665 
                 AACACUCUUCUUGAGCAGA 
                 149 
                 2665 
                 AACACUCUUCUUGAGCAGA 
                 149 
                 2683 
                 UCUGCUCAAGAAGAGUGUU 
                 334 
               
               
                   
               
               
                 2683 
                 ACCGUGAUUUGGAAGAGAG 
                 150 
                 2683 
                 ACCGUGAUUUGGAAGAGAG 
                 150 
                 2701 
                 CUCUCUUCCAAAUCACGGU 
                 335 
               
               
                   
               
               
                 2701 
                 GGCACCUGCUGGAAACCAC 
                 151 
                 2701 
                 GGCACCUGCUGGAAACCAC 
                 151 
                 2719 
                 GUGGUUUCCAGCAGGUGCC 
                 336 
               
               
                   
               
               
                 2719 
                 CACUUCUUGAAACAGCCUG 
                 152 
                 2719 
                 CACUUCUUGAAACAGCCUG 
                 152 
                 2737 
                 CAGGCUGUUUCAAGAAGUG 
                 337 
               
               
                   
               
               
                 2737 
                 GGGUGACGGUCCUUUAGGC 
                 153 
                 2737 
                 GGGUGACGGUCCUUUAGGC 
                 153 
                 2755 
                 GCCUAAAGGACCGUCACCC 
                 338 
               
               
                   
               
               
                 2755 
                 CAGCCUGCCGCCGUCUCUG 
                 154 
                 2755 
                 CAGCCUGCCGCCGUCUCUG 
                 154 
                 2773 
                 CAGAGACGGCGGCAGGCUG 
                 339 
               
               
                   
               
               
                 2773 
                 GUCCCGGUUCACCUUGCCG 
                 155 
                 2773 
                 GUCCCGGUUCACCUUGCCG 
                 155 
                 2791 
                 CGGCAAGGUGAACCGGGAC 
                 340 
               
               
                   
               
               
                 2791 
                 GAGAGAGGCGCGUGUGCCC 
                 156 
                 2791 
                 GAGAGAGGCGCGUCUGCCC 
                 156 
                 2809 
                 GGGCAGACGCGCCUCUCUC 
                 341 
               
               
                   
               
               
                 2809 
                 CCACCCUCAAACCCUGUGG 
                 157 
                 2809 
                 CCACCCUCAAACCCUGUGG 
                 157 
                 2827 
                 CCACAGGGUUUGAGGGUGG 
                 342 
               
               
                   
               
               
                 2827 
                 GGGCCUGAUGGUGCUCACG 
                 158 
                 2827 
                 GGGCCUGAUGGUGCUCACG 
                 158 
                 2845 
                 CGUGAGCACCAUCAGGCCC 
                 343 
               
               
                   
               
               
                 2845 
                 GACUCUUCCUGCAAAGGGA 
                 159 
                 2845 
                 GACUCUUCCUGCAAAGGGA 
                 159 
                 2863 
                 UCCCUUUGCAGGAAGAGUC 
                 344 
               
               
                   
               
               
                 2863 
                 AACUGAAGACCUCCACAUU 
                 160 
                 2863 
                 AACUGAAGACCUCCACAUU 
                 160 
                 2881 
                 AAUGUGGAGGUCUUCAGUU 
                 345 
               
               
                   
               
               
                 2881 
                 UAAGUGGCUUUUUAACAUG 
                 161 
                 2881 
                 UAAGUGGCUUUUUAACAUG 
                 161 
                 2899 
                 CAUGUUAAAAAGCCACUUA 
                 346 
               
               
                   
               
               
                 2899 
                 GAAAAACACGGCAGCUGUA 
                 162 
                 2899 
                 GAAAAACACGGCAGCUGUA 
                 162 
                 2917 
                 UACAGCUGCCGUGUUUUUC 
                 347 
               
               
                   
               
               
                 2917 
                 AGCUCCCGAGCUACUCUCU 
                 163 
                 2917 
                 AGCUCCCGAGCUACUCUCU 
                 163 
                 2935 
                 AGAGAGUAGCUCGGGAGCU 
                 348 
               
               
                   
               
               
                 2935 
                 UUGCCAGCAUUUUCACAUU 
                 164 
                 2935 
                 UUGCCAGCAUUUUCACAUU 
                 164 
                 2953 
                 AAUGUGAAAAUGCUGGCAA 
                 349 
               
               
                   
               
               
                 2953 
                 UUUGCCUUUCUCGUGGUAG 
                 165 
                 2953 
                 UUUGCCUUUCUCGUGGUAG 
                 165 
                 2971 
                 CUACCACGAGAAAGGCAAA 
                 350 
               
               
                   
               
               
                 2971 
                 GAAGCCAGUACAGAGAAAU 
                 166 
                 2971 
                 GAAGCCAGUACAGAGAAAU 
                 166 
                 2989 
                 AUUUCUCUGUACUGGCUUC 
                 351 
               
               
                   
               
               
                 2989 
                 UUCUGUGGUGGGAACAUUC 
                 167 
                 2989 
                 UUCUGUGGUGGGAACAUUC 
                 167 
                 3007 
                 GAAUGUUCCCACCACAGAA 
                 352 
               
               
                   
               
               
                 3007 
                 CGAGGUGUCACCCUGCAGA 
                 168 
                 3007 
                 CGAGGUGUCACCCUGCAGA 
                 168 
                 3025 
                 UCUGCAGGGUGACACCUCG 
                 353 
               
               
                   
               
               
                 3025 
                 AGCUAUGGUGAGGUGUGGA 
                 169 
                 3025 
                 AGCUAUGGUGAGGUGUGGA 
                 169 
                 3043 
                 UCCACACCUCACCAUAGCU 
                 354 
               
               
                   
               
               
                 3043 
                 AUAAGGCUUAGGUGCCAGG 
                 170 
                 3043 
                 AUAAGGCUUAGGUGCCAGG 
                 170 
                 3061 
                 CCUGGCACCUAAGCCUUAU 
                 355 
               
               
                   
               
               
                 3061 
                 GCUGUAAGCAUUCUGAGCU 
                 171 
                 3061 
                 GCUGUAAGCAUUCUGAGCU 
                 171 
                 3079 
                 AGCUCAGAAUGCUUACAGC 
                 356 
               
               
                   
               
               
                 3079 
                 UGGGCUUGUUGUUUUUAAG 
                 172 
                 3079 
                 UGGGCUUGUUGUUUUUAAG 
                 172 
                 3097 
                 CUUAAAAACAACAAGCCCA 
                 357 
               
               
                   
               
               
                 3097 
                 GUCCUGUAUAUGUAUGUAG 
                 173 
                 3097 
                 GUCCUGUAUAUGUAUGUAG 
                 173 
                 3115 
                 CUACAUACAUAUACAGGAC 
                 358 
               
               
                   
               
               
                 3115 
                 GUAGUUUGGGUGUGUAUAU 
                 174 
                 3115 
                 GUAGUUUGGGUGUGUAUAU 
                 174 
                 3133 
                 AUAUACACACCCAAACUAC 
                 359 
               
               
                   
               
               
                 3133 
                 UAUAGUAGCAUUUCAAAAU 
                 175 
                 3133 
                 UAUAGUAGCAUUUCAAAAU 
                 175 
                 3151 
                 AUUUUGAAAUGCUACUAUA 
                 360 
               
               
                   
               
               
                 3151 
                 UGGACGUACUGGUUUAACC 
                 176 
                 3151 
                 UGGACGUACUGGUUUAACC 
                 176 
                 3169 
                 GGUUAAACCAGUACGUCCA 
                 361 
               
               
                   
               
               
                 3169 
                 CUCCUAUCCUUGGAGAGCA 
                 177 
                 3169 
                 CUCCUAUCCUUGGAGAGCA 
                 177 
                 3187 
                 UGCUCUCCAAGGAUAGGAG 
                 362 
               
               
                   
               
               
                 3187 
                 AGCUGGCUCUCCACCUUGU 
                 178 
                 3187 
                 AGCUGGCUCUCCACCUUGU 
                 178 
                 3205 
                 ACAAGGUGGAGAGCCAGCU 
                 363 
               
               
                   
               
               
                 3205 
                 UUACACAUUAUGUUAGAGA 
                 179 
                 3205 
                 UUACACAUUAUGUUAGAGA 
                 179 
                 3223 
                 UCUCUAACAUAAUGUGUAA 
                 364 
               
               
                   
               
               
                 3223 
                 AGGUAGCGAGCUGCUCUGC 
                 180 
                 3223 
                 AGGUAGCGAGCUGCUCUGC 
                 180 
                 3241 
                 GCAGAGCAGCUCGCUACCU 
                 365 
               
               
                   
               
               
                 3241 
                 CUAUAUGCCUUAAGCCAAU 
                 181 
                 3241 
                 CUAUAUGCCUUAAGCCAAU 
                 181 
                 3259 
                 AUUGGCUUAAGGCAUAUAG 
                 366 
               
               
                   
               
               
                 3259 
                 UAUUUACUCAUCAGGUCAU 
                 182 
                 3259 
                 UAUUUACUCAUCAGGUCAU 
                 182 
                 3277 
                 AUGACCUGAUGAGUAAAUA 
                 367 
               
               
                   
               
               
                 3277 
                 UUAUUUUUUACAAUGGCCA 
                 183 
                 3277 
                 UUAUUUUUUACAAUGGCCA 
                 183 
                 3295 
                 UGGCCAUUGUAAAAAAUAA 
                 368 
               
               
                   
               
               
                 3295 
                 AUGGAAUAAACCAUUUUUA 
                 184 
                 3295 
                 AUGGAAUAAACCAUUUUUA 
                 184 
                 3313 
                 UAAAAAUGGUUUAUUCCAU 
                 369 
               
               
                   
               
               
                 3300 
                 AUAAACCAUUUUUACAAAA 
                 185 
                 3300 
                 AUAAACCAUUUUUACAAAA 
                 185 
                 3318 
                 UUUUGUAAAAAUGGUUUAU 
                 370 
               
               
                   
               
               
                 The 3′-ends of the Upper sequence and the Lower sequence of the siNA construct can include an overhang sequence, for example about 1, 2, 3, or 4 nucleotides in length, preferably 2 nucleotides in length, wherein the overhanging sequence of the lower sequence is optionally complementary to a portion of the target sequence. The upper sequence is also referred to as the sense strand, whereas the lower sequence is also referred to as the antisense strand. The upper and lower sequences in the Table can further comprise a chemical modification having Formulae I-VII, such as exemplary siNA constructs shown in FIGS. 4 and 5, or having modifications described in Table IV or any combination thereof. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE III 
               
               
                   
               
               
                 PTP-1B Synthetic Modified siNA constructs 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 Target 
                   
                 Seq 
                   
                   
                   
               
               
                 Pos 
                 Target 
                 ID 
                 Cmpd# 
                 Aliases 
               
               
                   
               
               
                  240 
                 UAUCCGACAUGAAGCCAGUGACU 
                 371 
                 31017 
                 PTPN1:242U21 sense siNA 
               
               
                   
               
               
                  282 
                 GCUUCCUAAGAACAAAAACCGAA 
                 372 
                   
                 PTPN1:284U21 sense siNA 
               
               
                   
               
               
                  356 
                 AAGAAGAUAAUGACUAUAUCAAC 
                 373 
                   
                 PTPN1:358U21 sense siNA 
               
               
                   
               
               
                  357 
                 AGAAGAUAAUGACUAUAUCAACG 
                 374 
                   
                 PTPN1:359U21 sense siNA 
               
               
                   
               
               
                  458 
                 UUUGGGAGAUGGUGUGGGAGCAG 
                 375 
                   
                 PTPN1:460U21 sense siNA 
               
               
                   
               
               
                  460 
                 UGGGAGAUGGUGUGGGAGCAGAA 
                 376 
                   
                 PTPN1:462U21 sense siNA 
               
               
                   
               
               
                  708 
                 AUGGCCUGACUUUGGAGUCCCUG 
                 377 
                   
                 PTPN1:710U21 sense siNA 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                   
                 PTPN1:761U21 sense siNA 
               
               
                   
               
               
                  764 
                 AAGUCCGAGAGUCAGGGUCACUC 
                 379 
                 31018 
                 PTPN1:766U21 sense siNA 
               
               
                   
               
               
                  872 
                 UGCUGAUGGACAAGAGGAAAGAC 
                 380 
                 31019 
                 PTPN1:874U21 sense siNA 
               
               
                   
               
               
                  874 
                 CUGAUGGACAAGAGGAAAGACCC 
                 381 
                   
                 PTPN1:876U21 sense siNA 
               
               
                   
               
               
                  875 
                 UGAUGGACAAGAGGAAAGACCCU 
                 382 
                   
                 PTPN1:877U21 sense siNA 
               
               
                   
               
               
                  876 
                 GAUGGACAAGAGGAAAGACCCUU 
                 383 
                   
                 PTPN1:878U21 sense siNA 
               
               
                   
               
               
                  973 
                 CUGCGCUUCUCCUACCUGGCUGU 
                 384 
                   
                 PTPN1:975U21 sense siNA 
               
               
                   
               
               
                 1033 
                 GUGCAGGAUCAGUGGAAGGAGCU 
                 385 
                   
                 PTPN1:1035U21 sense siNA 
               
               
                   
               
               
                 1035 
                 GCAGGAUCAGUGGAAGGAGCUUU 
                 386 
                   
                 PTPN1:1037U21 sense siNA 
               
               
                   
               
               
                 1094 
                 CCCCACCUCCCCGGCCACCCAAA 
                 387 
                   
                 PTPN1:1096U21 sense siNA 
               
               
                   
               
               
                 1386 
                 UUACUGGAAGCCCUUCCUGGUCA 
                 388 
                   
                 PTPN1:1388U21 sense siNA 
               
               
                   
               
               
                 3035 
                 AGGUGUGGAUAAGGCUUAGGUGC 
                 389 
                 31020 
                 PTPN1:3037U21 sense siNA 
               
               
                   
               
               
                  240 
                 UAUCCGACAUGAAGCCAGUGACU 
                 371 
                 31093 
                 PTPN1:260L21 sense siNA (242C) 
               
               
                   
               
               
                  282 
                 GCUUCCUAAGAACAAAAACCGAA 
                 372 
                   
                 PTPN1:302L21 sense siNA (284C) 
               
               
                   
               
               
                  356 
                 AAGAAGAUAAUGACUAUAUCAAC 
                 373 
                   
                 PTPN1:376L21 sense siNA (358C) 
               
               
                   
               
               
                  357 
                 AGAAGAUAAUGACUAUAUCAACG 
                 374 
                   
                 PTPN1:377L21 sense siNA (359C) 
               
               
                   
               
               
                  458 
                 UUUGGGAGAUGGUGUGGGAGCAG 
                 375 
                   
                 PTPN1:478L21 sense siNA (460C) 
               
               
                   
               
               
                  460 
                 UGGGAGAUGGUGUGGGAGCAGAA 
                 376 
                   
                 PTPN1:480L21 sense siNA (462C) 
               
               
                   
               
               
                  708 
                 AUGGCCUGACUUUGGAGUCCCUG 
                 377 
                   
                 PTPN1:728L21 sense siNA (710C) 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                   
                 PTPN1:779L21 sense siNA (761C) 
               
               
                   
               
               
                  764 
                 AAGUCCGAGAGUCAGGGUCACUC 
                 379 
                 31094 
                 PTPN1:784L21 sense siNA (766C) 
               
               
                   
               
               
                  872 
                 UGCUGAUGGACAAGAGGAAAGAC 
                 380 
                 31095 
                 PTPN1:892L21 sense siNA (874C) 
               
               
                   
               
               
                  874 
                 CUGAUGGACAAGAGGAAAGACCC 
                 381 
                   
                 PTPN1:894L21 sense siNA (876C) 
               
               
                   
               
               
                  875 
                 UGAUGGACAAGAGGAAAGACCCU 
                 382 
                   
                 PTPN1:895L21 sense siNA (877C) 
               
               
                   
               
               
                  876 
                 GAUGGACAAGAGGAAAGACCCUU 
                 383 
                   
                 PTPN1:896L21 sense siNA (878C) 
               
               
                   
               
               
                  973 
                 CUGCGCUUCUCCUACCUGGCUGU 
                 384 
                   
                 PTPN1:993L21 sense siNA (975C) 
               
               
                   
               
               
                 1033 
                 GUGCAGGAUCAGUGGAAGGAGCU 
                 385 
                   
                 PTPN1:1053L21 sense siNA (1035C) 
               
               
                   
               
               
                 1035 
                 GCAGGAUCAGUGGAAGGAGCUUU 
                 386 
                   
                 PTPN1:1055L21 sense siNA (1037C) 
               
               
                   
               
               
                 1094 
                 CCCCACCUCCCCGGCCACCCAAA 
                 387 
                   
                 PTPN1:1114L21 sense siNA (1096C) 
               
               
                   
               
               
                 1386 
                 UUACUGGAAGCCCUUCCUGGUCA 
                 388 
                   
                 PTPN1:1406L21 sense siNA (1388C) 
               
               
                   
               
               
                 3035 
                 AGGUGUGGAUAAGGCUUAGGUGC 
                 389 
                 31096 
                 PTPN1:3055L21 sense siNA (3037C) 
               
               
                   
               
               
                  240 
                 UAUCCGACAUGAAGCCAGUGACU 
                 371 
                 30865 
                 PTPN1:242U21 sense siNA stab04 
               
               
                   
               
               
                  282 
                 GCUUCCUAAGAACAAAAACCGAA 
                 372 
                 39283 
                 PTPN1:284U21 sense siNA stab04 
               
               
                   
               
               
                  356 
                 AAGAAGAUAAUGACUAUAUCAAC 
                 373 
                 39284 
                 PTPN1:358U21 sense siNA stab04 
               
               
                   
               
               
                  357 
                 AGAAGAUAAUGACUAUAUCAACG 
                 374 
                 39285 
                 PTPN1:359U21 sense siNA stab04 
               
               
                   
               
               
                  458 
                 UUUGGGAGAUGGUGUGGGAGCAG 
                 375 
                 39286 
                 PTPN1:460U21 sense siNA stab04 
               
               
                   
               
               
                  460 
                 UGGGAGAUGGUGUGGGAGCAGAA 
                 376 
                 39287 
                 PTPN1:462U21 sense siNA stab04 
               
               
                   
               
               
                  708 
                 AUGGCCUGACUUUGGAGUCCCUG 
                 377 
                 39288 
                 PTPN1:710U21 sense siNA stab04 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                 39289 
                 PTPN1:761U21 sense siNA stab04 
               
               
                   
               
               
                  764 
                 AAGUCCGAGAGUCAGGGUCACUC 
                 379 
                 31306 
                 PTPN1:766U21 sense siNA stab04 
               
               
                   
               
               
                  872 
                 UGCUGAUGGACAAGAGGAAAGAC 
                 380 
                 39290 
                 PTPN1:874U21 sense siNA stab04 
               
               
                   
               
               
                  874 
                 CUGAUGGACAAGAGGAAAGACCC 
                 381 
                 39291 
                 PTPN1:876U21 sense siNA stab04 
               
               
                   
               
               
                  875 
                 UGAUGGACAAGAGGAAAGACCCU 
                 382 
                 39292 
                 PTPN1:877U21 sense siNA stab04 
               
               
                   
               
               
                  876 
                 GAUGGACAAGAGGAAAGACCCUU 
                 383 
                 39293 
                 PTPN1:878U21 sense siNA stab04 
               
               
                   
               
               
                  973 
                 CUGCGCUUCUCCUACCUGGCUGU 
                 384 
                 39294 
                 PTPN1:975U21 sense siNA stab04 
               
               
                   
               
               
                 1033 
                 GUGCAGGAUCAGUGGAAGGAGCU 
                 385 
                 39295 
                 PTPN1:1035U21 sense siNA stab04 
               
               
                   
               
               
                 1035 
                 GCAGGAUCAGUGGAAGGAGCUUU 
                 386 
                 39296 
                 PTPN1:1037U21 sense siNA stab04 
               
               
                   
               
               
                 1094 
                 CCCCACCUCCCCGGCCACCCAAA 
                 387 
                 39297 
                 PTPN1:1096U21 sense siNA stab04 
               
               
                   
               
               
                 1386 
                 UUACUGGAAGCCCUUCCUGGUCA 
                 388 
                 39298 
                 PTPN1:1388U21 sense siNA stab04 
               
               
                   
               
               
                 3035 
                 AGGUGUGGAUAAGGCUUAGGUGC 
                 389 
                 30868 
                 PTPN1:3037U21 sense siNA stab04 
               
               
                   
               
               
                  240 
                 UAUCCGACAUGAAGCCAGUGACU 
                 371 
                 30869 
                 PTPN1:260L21 sense siNA (242C) stab05 
               
               
                   
               
               
                  282 
                 GCUUCCUAAGAACAAAAACCGAA 
                 372 
                   
                 PTPN1:302L21 sense siNA (284C) stab05 
               
               
                   
               
               
                  356 
                 AAGAAGAUAAUGACUAUAUCAAC 
                 373 
                   
                 PTPN1:376L21 sense siNA (358C) stab05 
               
               
                   
               
               
                  357 
                 AGAAGAUAAUGACUAUAUCAACG 
                 374 
                   
                 PTPN1:377L21 sense siNA (359C) stab05 
               
               
                   
               
               
                  458 
                 UUUGGGAGAUGGUGUGGGAGCAG 
                 375 
                   
                 PTPN1:478L21 sense siNA (460C) stab05 
               
               
                   
               
               
                  460 
                 UGGGAGAUGGUGUGGGAGCAGAA 
                 376 
                   
                 PTPN1:480L21 sense siNA (462C) stab05 
               
               
                   
               
               
                  708 
                 AUGGCCUGACUUUGGAGUCCCUG 
                 377 
                   
                 PTPN1:728L21 sense siNA (710C) stab05 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                   
                 PTPN1:779L21 sense siNA (761C) stab05 
               
               
                   
               
               
                  764 
                 AAGUCCGAGAGUCAGGGUCACUC 
                 379 
                 31307 
                 PTPN1:784L21 sense siNA (766C) stab05 
               
               
                   
               
               
                  872 
                 UGCUGAUGGACAAGAGGAAAGAC 
                 380 
                 30871 
                 PTPN1:892L21 sense siNA (874C) stab05 
               
               
                   
               
               
                  874 
                 CUGAUGGACAAGAGGAAAGACCC 
                 381 
                   
                 PTPN1:894L21 sense siNA (876C) stab05 
               
               
                   
               
               
                  875 
                 UGAUGGACAAGAGGAAAGACCCU 
                 382 
                   
                 PTPN1:895L21 sense siNA (877C) stab05 
               
               
                   
               
               
                  876 
                 GAUGGACAAGAGGAAAGACCCUU 
                 383 
                   
                 PTPN1:896L21 sense siNA (878C) stab05 
               
               
                   
               
               
                  973 
                 CUGCGCUUCUCCUACCUGGCUGU 
                 384 
                   
                 PTPN1:993L21 sense siNA (975C) stab05 
               
               
                   
               
               
                 1033 
                 GUGCAGGAUCAGUGGAAGGAGCU 
                 385 
                   
                 PTPN1:1053L21 sense siNA (1035C) stab05 
               
               
                   
               
               
                 1035 
                 GCAGGAUCAGUGGAAGGAGCUUU 
                 386 
                   
                 PTPN1:1055L21 sense siNA (1037C) stab05 
               
               
                   
               
               
                 1094 
                 CCCCACCUCCCCGGCCACCCAAA 
                 387 
                   
                 PTPN1:1114L21 sense siNA (1096C) stab05 
               
               
                   
               
               
                 1386 
                 UUACUGGAAGCCCUUCCUGGUCA 
                 388 
                   
                 PTPN1:1406L21 sense siNA (1388C) stab05 
               
               
                   
               
               
                 3035 
                 AGGUGUGGAUAAGGCUUAGGUGC 
                 389 
                 30872 
                 PTPN1:3055L21 sense siNA (3037C) stab05 
               
               
                   
               
               
                  240 
                 UAUCCGACAUGAAGCCAGUGACU 
                 371 
                   
                 PTPN1:242U21 sense siNA stab07 
               
               
                   
               
               
                  282 
                 GCUUCCUAAGAACAAAAACCGAA 
                 372 
                 39299 
                 PTPN1:284U21 sense siNA stab07 
               
               
                   
               
               
                  356 
                 AAGAAGAUAAUGACUAUAUCAAC 
                 373 
                 39300 
                 PTPN1:358U21 sense siNA stab07 
               
               
                   
               
               
                  357 
                 AGAAGAUAAUGACUAUAUCAACG 
                 374 
                 39301 
                 PTPN1:359U21 sense siNA stab07 
               
               
                   
               
               
                  458 
                 UUUGGGAGAUGGUGUGGGAGCAG 
                 375 
                 39302 
                 PTPN1:460U21 sense siNA stab07 
               
               
                   
               
               
                  460 
                 UGGGAGAUGGUGUGGGAGCAGAA 
                 376 
                 39303 
                 PTPN1:462U21 sense siNA stab07 
               
               
                   
               
               
                  708 
                 AUGGCCUGACUUUGGAGUCCCUG 
                 377 
                 39304 
                 PTPN1:710U21 sense siNA stab07 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                 39305 
                 PTPN1:761U21 sense siNA stab07 
               
               
                   
               
               
                  764 
                 AAGUCCGAGAGUCAGGGUCACUC 
                 379 
                   
                 PTPN1:766U21 sense siNA stab07 
               
               
                   
               
               
                  872 
                 UGCUGAUGGACAAGAGGAAAGAC 
                 380 
                 39306 
                 PTPN1:874U21 sense siNA stab07 
               
               
                   
               
               
                  874 
                 CUGAUGGACAAGAGGAAAGACCC 
                 381 
                 39307 
                 PTPN1:876U21 sense siNA stab07 
               
               
                   
               
               
                  875 
                 UGAUGGACAAGAGGAAAGACCCU 
                 382 
                 39308 
                 PTPN1:877U21 sense siNA stab07 
               
               
                   
               
               
                  876 
                 GAUGGACAAGAGGAAAGACCCUU 
                 383 
                 39309 
                 PTPN1:878U21 sense siNA stab07 
               
               
                   
               
               
                  973 
                 CUGCGCUUCUCCUACCUGGCUGU 
                 384 
                 39310 
                 PTPN1:975U21 sense siNA stab07 
               
               
                   
               
               
                 1033 
                 GUGCAGGAUCAGUGGAAGGAGCU 
                 385 
                 39311 
                 PTPN1:1035U21 sense siNA stab07 
               
               
                   
               
               
                 1035 
                 GCAGGAUCAGUGGAAGGAGCUUU 
                 386 
                 39312 
                 PTPN1:1037U21 sense siNA stab07 
               
               
                   
               
               
                 1094 
                 CCCCACCUCCCCGGCCACCCAAA 
                 387 
                 39313 
                 PTPN1:1096U21 sense siNA stab07 
               
               
                   
               
               
                 1386 
                 UUACUGGAAGCCCUUCCUGGUCA 
                 388 
                 39314 
                 PTPN1:1388U21 sense siNA stab07 
               
               
                   
               
               
                 3035 
                 AGGUGUGGAUAAGGCUUAGGUGC 
                 389 
                   
                 PTPN1:3037U21 sense siNA stab07 
               
               
                   
               
               
                  240 
                 UAUCCGACAUGAAGCCAGUGACU 
                 371 
                   
                 PTPN1:260L21 sense siNA (242C) stab11 
               
               
                   
               
               
                  282 
                 GCUUCCUAAGAACAAAAACCGAA 
                 372 
                   
                 PTPN1:302L21 sense siNA (284C) stab11 
               
               
                   
               
               
                  356 
                 AAGAAGAUAAUGACUAUAUCAAC 
                 373 
                   
                 PTPN1:376L21 sense siNA (358C) stab11 
               
               
                   
               
               
                  357 
                 AGAAGAUAAUGACUAUAUCAACG 
                 374 
                   
                 PTPN1:377L21 sense siNA (359C) stab11 
               
               
                   
               
               
                  458 
                 UUUGGGAGAUGGUGUGGGAGCAG 
                 375 
                   
                 PTPN1:478L21 sense siNA (460C) stab11 
               
               
                   
               
               
                  460 
                 UGGGAGAUGGUGUGGGAGCAGAA 
                 376 
                   
                 PTPN1:480L21 sense siNA (462C) stab11 
               
               
                   
               
               
                  708 
                 AUGGCCUGACUUUGGAGUCCCUG 
                 377 
                   
                 PTPN1:728L21 sense siNA (710C) stab11 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                   
                 PTPN1:779L21 sense siNA (761C) stab11 
               
               
                   
               
               
                  764 
                 AAGUCCGAGAGUCAGGGUCACUC 
                 379 
                   
                 PTPN1:784L21 sense siNA (766C) stab11 
               
               
                   
               
               
                  872 
                 UGCUGAUGGACAAGAGGAAAGAC 
                 380 
                   
                 PTPN1:892L21 sense siNA (874C) stab11 
               
               
                   
               
               
                  874 
                 CUGAUGGACAAGAGGAAAGACCC 
                 381 
                   
                 PTPN1:894L21 sense siNA (876C) stab11 
               
               
                   
               
               
                  875 
                 UGAUGGACAAGAGGAAAGACCCU 
                 382 
                   
                 PTPN1:895L21 sense siNA (877C) stab11 
               
               
                   
               
               
                  876 
                 GAUGGACAAGAGGAAAGACCCUU 
                 383 
                   
                 PTPN1:896L21 sense siNA (878C) stab11 
               
               
                   
               
               
                  973 
                 CUGCGCUUCUCCUACCUGGCUGU 
                 384 
                   
                 PTPN1:993L21 sense siNA (975C) stab11 
               
               
                   
               
               
                 1033 
                 GUGCAGGAUCAGUGGAAGGAGCU 
                 385 
                   
                 PTPN1:1053L21 sense siNA (1035C) stab11 
               
               
                   
               
               
                 1035 
                 GCAGGAUCAGUGGAAGGAGCUUU 
                 386 
                   
                 PTPN1:1055L21 sense siNA (1037C) stab11 
               
               
                   
               
               
                 1094 
                 CCCCACCUCCCCGGCCACCCAAA 
                 387 
                   
                 PTPN1:1114L21 sense siNA (1096C) stab11 
               
               
                   
               
               
                 1386 
                 UUACUGGAAGCCCUUCCUGGUCA 
                 388 
                   
                 PTPN1:1406L21 sense siNA (1388C) stab11 
               
               
                   
               
               
                 3035 
                 AGGUGUGGAUAAGGCUUAGGUGC 
                 389 
                   
                 PTPN1:3055L21 sense siNA (3037C) stab11 
               
               
                   
               
               
                  240 
                 UAUCCGACAUGAAGCCAGUGACU 
                 371 
                   
                 PTPN1:242U21 sense siNA stab18 
               
               
                   
               
               
                  282 
                 GCUUCCUAAGAACAAAAACCGAA 
                 372 
                   
                 PTPN1:284U21 sense siNA stab18 
               
               
                   
               
               
                  356 
                 AAGAAGAUAAUGACUAUAUCAAC 
                 373 
                   
                 PTPN1:358U21 sense siNA stab18 
               
               
                   
               
               
                  357 
                 AGAAGAUAAUGACUAUAUCAACG 
                 374 
                   
                 PTPN1:359U21 sense siNA stab18 
               
               
                   
               
               
                  458 
                 UUUGGGAGAUGGUGUGGGAGCAG 
                 375 
                   
                 PTPN1:460U21 sense siNA stab18 
               
               
                   
               
               
                  460 
                 UGGGAGAUGGUGUGGGAGCAGAA 
                 376 
                   
                 PTPN1:462U21 sense siNA stab18 
               
               
                   
               
               
                  708 
                 AUGGCCUGACUUUGGAGUCCCUG 
                 377 
                   
                 PTPN1:710U21 sense siNA stab18 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                   
                 PTPN1:761U21 sense siNA stab18 
               
               
                   
               
               
                  764 
                 AAGUCCGAGAGUCAGGGUCACUC 
                 379 
                   
                 PTPN1:766U21 sense siNA stab18 
               
               
                   
               
               
                  872 
                 UGCUGAUGGACAAGAGGAAAGAC 
                 380 
                   
                 PTPN1:874U21 sense siNA stab18 
               
               
                   
               
               
                  874 
                 CUGAUGGACAAGAGGAAAGACCC 
                 381 
                   
                 PTPN1:876U21 sense siNA stab18 
               
               
                   
               
               
                  875 
                 UGAUGGACAAGAGGAAAGACCCU 
                 382 
                   
                 PTPN1:877U21 sense siNA stab18 
               
               
                   
               
               
                  876 
                 GAUGGACAAGAGGAAAGACCCUU 
                 383 
                   
                 PTPN1:878U21 sense siNA stab18 
               
               
                   
               
               
                  973 
                 CUGCGCUUCUCCUACCUGGCUGU 
                 384 
                   
                 PTPN1:975U21 sense siNA stab18 
               
               
                   
               
               
                 1033 
                 GUGCAGGAUCAGUGGAAGGAGCU 
                 385 
                   
                 PTPN1:1035U21 sense siNA stab18 
               
               
                   
               
               
                 1035 
                 GCAGGAUCAGUGGAAGGAGCUUU 
                 386 
                   
                 PTPN1:1037U21 sense siNA stab18 
               
               
                   
               
               
                 1094 
                 CCCCACCUCCCCGGCCACCCAAA 
                 387 
                   
                 PTPN1:1096U21 sense siNA stab18 
               
               
                   
               
               
                 1386 
                 UUACUGGAAGCCCUUCCUGGUCA 
                 388 
                   
                 PTPN1:1388U21 sense siNA stab18 
               
               
                   
               
               
                 3035 
                 AGGUGUGGAUAAGGCUUAGGUGC 
                 389 
                   
                 PTPN1:3037U21 sense siNA stab18 
               
               
                   
               
               
                  240 
                 UAUCCGACAUGAAGCCAGUGACU 
                 371 
                   
                 PTPN1:260L21 sense siNA (242C) stab08 
               
               
                   
               
               
                  282 
                 GCUUCCUAAGAACAAAAACCGAA 
                 372 
                   
                 PTPN1:302L21 sense siNA (284C) stab08 
               
               
                   
               
               
                  356 
                 AAGAAGAUAAUGACUAUAUCAAC 
                 373 
                   
                 PTPN1:376L21 sense siNA (358C) stab08 
               
               
                   
               
               
                  357 
                 AGAAGAUAAUGACUAUAUCAACG 
                 374 
                   
                 PTPN1:377L21 sense siNA (359C) stab08 
               
               
                   
               
               
                  458 
                 UUUGGGAGAUGGUGUGGGAGCAG 
                 375 
                   
                 PTPN1:478L21 sense siNA (460C) stab08 
               
               
                   
               
               
                  460 
                 UGGGAGAUGGUGUGGGAGCAGAA 
                 376 
                   
                 PTPN1:480L21 sense siNA (462C) stab08 
               
               
                   
               
               
                  708 
                 AUGGCCUGACUUUGGAGUCCCUG 
                 377 
                   
                 PTPN1:728L21 sense siNA (710C) stab08 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                 39709 
                 PTPN1:779L21 sense siNA (761C) stab08 
               
               
                   
               
               
                  764 
                 AAGUCCGAGAGUCAGGGUCACUC 
                 379 
                   
                 PTPN1:784L21 sense siNA (766C) stab08 
               
               
                   
               
               
                  872 
                 UGCUGAUGGACAAGAGGAAAGAC 
                 380 
                   
                 PTPN1:892L21 sense siNA (874C) stab08 
               
               
                   
               
               
                  874 
                 CUGAUGGACAAGAGGAAAGACCC 
                 381 
                   
                 PTPN1:894L21 sense siNA (876C) stab08 
               
               
                   
               
               
                  875 
                 UGAUGGACAAGAGGAAAGACCCU 
                 382 
                   
                 PTPN1:895L21 sense siNA (877C) stab08 
               
               
                   
               
               
                  876 
                 GAUGGACAAGAGGAAAGACCCUU 
                 383 
                   
                 PTPN1:896L21 sense siNA (878C) stab08 
               
               
                   
               
               
                  973 
                 CUGCGCUUCUCCUACCUGGCUGU 
                 384 
                   
                 PTPN1:993L21 sense siNA (975C) stab08 
               
               
                   
               
               
                 1033 
                 GUGCAGGAUCAGUGGAAGGAGCU 
                 385 
                   
                 PTPN1:1053L21 sense siNA (1035C) stab08 
               
               
                   
               
               
                 1035 
                 GCAGGAUCAGUGGAAGGAGCUUU 
                 386 
                   
                 PTPN1:1055L21 sense siNA (1037C) stab08 
               
               
                   
               
               
                 1094 
                 CCCCACCUCCCCGGCCACCCAAA 
                 387 
                   
                 PTPN1:1114L21 sense siNA (1096C) stab08 
               
               
                   
               
               
                 1386 
                 UUACUGGAAGCCCUUCCUGGUCA 
                 388 
                   
                 PTPN1:1406L21 sense siNA (1388C) stab08 
               
               
                   
               
               
                 3035 
                 AGGUGUGGAUAAGGCUUAGGUGC 
                 389 
                   
                 PTPN1:3055L21 sense siNA (3037C) stab08 
               
               
                   
               
               
                  240 
                 UAUCCGACAUGAAGCCAGUGACU 
                 371 
                   
                 PTPN1:242U21 sense siNA stab09 
               
               
                   
               
               
                  282 
                 GCUUCCUAAGAACAAAAACCGAA 
                 372 
                 39251 
                 PTPN1:284U21 sense siNA stab09 
               
               
                   
               
               
                  356 
                 AAGAAGAUAAUGACUAUAUCAAC 
                 373 
                 39252 
                 PTPN1:358U21 sense siNA stab09 
               
               
                   
               
               
                  357 
                 AGAAGAUAAUGACUAUAUCAACG 
                 374 
                 39253 
                 PTPN1:359U21 sense siNA stab09 
               
               
                   
               
               
                  458 
                 UUUGGGAGAUGGUGUGGGAGCAG 
                 375 
                 39254 
                 PTPN1:460U21 sense siNA stab09 
               
               
                   
               
               
                  460 
                 UGGGAGAUGGUGUGGGAGCAGAA 
                 376 
                 39255 
                 PTPN1:462U21 sense siNA stab09 
               
               
                   
               
               
                  708 
                 AUGGCCUGACUUUGGAGUCCCUG 
                 377 
                 39256 
                 PTPN1:710U21 sense siNA stab09 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                 39257 
                 PTPN1:761U21 sense siNA stab09 
               
               
                   
               
               
                  764 
                 AAGUCCGAGAGUCAGGGUCACUC 
                 379 
                   
                 PTPN1:766U21 sense siNA stab09 
               
               
                   
               
               
                  872 
                 UGCUGAUGGACAAGAGGAAAGAC 
                 380 
                 39258 
                 PTPN1:874U21 sense siNA stab09 
               
               
                   
               
               
                  874 
                 CUGAUGGACAAGAGGAAAGACCC 
                 381 
                 39259 
                 PTPN1:876U21 sense siNA stab09 
               
               
                   
               
               
                  875 
                 UGAUGGACAAGAGGAAAGACCCU 
                 382 
                 39260 
                 PTPN1:877U21 sense siNA stab09 
               
               
                   
               
               
                  876 
                 GAUGGACAAGAGGAAAGACCCUU 
                 383 
                 39261 
                 PTPN1:878U21 sense siNA stab09 
               
               
                   
               
               
                  973 
                 CUGCGCUUCUCCUACCUGGCUGU 
                 384 
                 39262 
                 PTPN1:975U21 sense siNA stab09 
               
               
                   
               
               
                 1033 
                 GUGCAGGAUCAGUGGAAGGAGCU 
                 385 
                 39263 
                 PTPN1:1035U21 sense siNA stab09 
               
               
                   
               
               
                 1035 
                 GCAGGAUCAGUGGAAGGAGCUUU 
                 386 
                 39264 
                 PTPN1:1037U21 sense siNA stab09 
               
               
                   
               
               
                 1094 
                 CCCCACCUCCCCGGCCACCCAAA 
                 387 
                 39265 
                 PTPN1:1096U21 sense siNA stab09 
               
               
                   
               
               
                 1386 
                 UUACUGGAAGCCCUUCCUGGUCA 
                 388 
                 39266 
                 PTPN1:1388U21 sense siNA stab09 
               
               
                   
               
               
                 3035 
                 AGGUGUGGAUAAGGCUUAGGUGC 
                 389 
                   
                 PTPN1:3037U21 sense siNA stab09 
               
               
                   
               
               
                  240 
                 UAUCCGACAUGAAGCCAGUGACU 
                 371 
                   
                 PTPN1:260L21 sense siNA (242C) stab10 
               
               
                   
               
               
                  282 
                 GCUUCCUAAGAACAAAAACCGAA 
                 372 
                 39267 
                 PTPN1:302L21 sense siNA (284C) stab10 
               
               
                   
               
               
                  356 
                 AAGAAGAUAAUGACUAUAUCAAC 
                 373 
                 39268 
                 PTPN1:376L21 sense siNA (358C) stab10 
               
               
                   
               
               
                  357 
                 AGAAGAUAAUGACUAUAUCAACG 
                 374 
                 39269 
                 PTPN1:377L21 sense siNA (359C) stab10 
               
               
                   
               
               
                  458 
                 UUUGGGAGAUGGUGUGGGAGCAG 
                 375 
                 39270 
                 PTPN1:478L21 sense siNA (460C) stab10 
               
               
                   
               
               
                  460 
                 UGGGAGAUGGUGUGGGAGCAGAA 
                 376 
                 39271 
                 PTPN1:480L21 sense siNA (462C) stab10 
               
               
                   
               
               
                  708 
                 AUGGCCUGACUUUGGAGUCCCUG 
                 377 
                 39272 
                 PTPN1:728L21 sense siNA (710C) stab10 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                 39273 
                 PTPN1:779L21 sense siNA (761C) stab10 
               
               
                   
               
               
                  764 
                 AAGUCCGAGAGUCAGGGUCACUC 
                 379 
                   
                 PTPN1:784L21 sense siNA (766C) stab10 
               
               
                   
               
               
                  872 
                 UGCUGAUGGACAAGAGGAAAGAC 
                 380 
                 39274 
                 PTPN1:892L21 sense siNA (874C) stab10 
               
               
                   
               
               
                  874 
                 CUGAUGGACAAGAGGAAAGACCC 
                 381 
                 39275 
                 PTPN1:894L21 sense siNA (876C) stab10 
               
               
                   
               
               
                  875 
                 UGAUGGACAAGAGGAAAGACCCU 
                 382 
                 39276 
                 PTPN1:895L21 sense siNA (877C) stab10 
               
               
                   
               
               
                  876 
                 GAUGGACAAGAGGAAAGACCCUU 
                 383 
                 39277 
                 PTPN1:896L21 sense siNA (878C) stab10 
               
               
                   
               
               
                  973 
                 CUGCGCUUCUCCUACCUGGCUGU 
                 384 
                 39278 
                 PTPN1:993L21 sense siNA (975C) stab10 
               
               
                   
               
               
                 1033 
                 GUGCAGGAUCAGUGGAAGGAGCU 
                 385 
                 39279 
                 PTPN1:1053L21 sense siNA (1035C) stab10 
               
               
                   
               
               
                 1035 
                 GCAGGAUCAGUGGAAGGAGCUUU 
                 386 
                 39280 
                 PTPN1:1055L21 sense siNA (1037C) stab10 
               
               
                   
               
               
                 1094 
                 CCCCACCUCCCCGGCCACCCAAA 
                 387 
                 39281 
                 PTPN1:1114L21 sense siNA (1096C) stab10 
               
               
                   
               
               
                 1386 
                 UUACUGGAAGCCCUUCCUGGUCA 
                 388 
                 39282 
                 PTPN1:1406L21 sense siNA (1388C) stab10 
               
               
                   
               
               
                 3035 
                 AGGUGUGGAUAAGGCUUAGGUGC 
                 389 
                   
                 PTPN1:3055L21 sense siNA (3037C) stab10 
               
               
                   
               
               
                  240 
                 UAUCCGACAUGAAGCCAGUGACU 
                 371 
                   
                 PTPN1:260L21 sense siNA (242C) stab19 
               
               
                   
               
               
                  282 
                 GCUUCCUAAGAACAAAAACCGAA 
                 372 
                   
                 PTPN1:302L21 sense siNA (284C) stab19 
               
               
                   
               
               
                  356 
                 AAGAAGAUAAUGACUAUAUCAAC 
                 373 
                   
                 PTPN1:376L21 sense siNA (358C) stab19 
               
               
                   
               
               
                  357 
                 AGAAGAUAAUGACUAUAUCAACG 
                 374 
                   
                 PTPN1:377L21 sense siNA (359C) stab19 
               
               
                   
               
               
                  458 
                 UUUGGGAGAUGGUGUGGGAGCAG 
                 375 
                   
                 PTPN1:478L21 sense siNA (460C) stab19 
               
               
                   
               
               
                  460 
                 UGGGAGAUGGUGUGGGAGCAGAA 
                 376 
                   
                 PTPN1:480L21 sense siNA (462C) stab19 
               
               
                   
               
               
                  708 
                 AUGGCCUGACUUUGGAGUCCCUG 
                 377 
                   
                 PTPN1:728L21 sense siNA (710C) stab19 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                   
                 PTPN1:779L21 sense siNA (761C) stab19 
               
               
                   
               
               
                  764 
                 AAGUCCGAGAGUCAGGGUCACUC 
                 379 
                   
                 PTPN1:784L21 sense siNA (766C) stab19 
               
               
                   
               
               
                  872 
                 UGCUGAUGGACAAGAGGAAAGAC 
                 380 
                   
                 PTPN1:892L21 sense siNA (874C) stab19 
               
               
                   
               
               
                  874 
                 CUGAUGGACAAGAGGAAAGACCC 
                 381 
                   
                 PTPN1:894L21 sense siNA (876C) stab19 
               
               
                   
               
               
                  875 
                 UGAUGGACAAGAGGAAAGACCCU 
                 382 
                   
                 PTPN1:895L21 sense siNA (877C) stab19 
               
               
                   
               
               
                  876 
                 GAUGGACAAGAGGAAAGACCCUU 
                 383 
                   
                 PTPN1:896L21 sense siNA (878C) stab19 
               
               
                   
               
               
                  973 
                 CUGCGCUUCUCCUACCUGGCUGU 
                 384 
                   
                 PTPN1:993L21 sense siNA (975C) stab19 
               
               
                   
               
               
                 1033 
                 GUGCAGGAUCAGUGGAAGGAGCU 
                 385 
                   
                 PTPN1:1053L21 sense siNA (1035C) stab19 
               
               
                   
               
               
                 1035 
                 GCAGGAUCAGUGGAAGGAGCUUU 
                 386 
                   
                 PTPN1:1055L21 sense siNA (1037C) stab19 
               
               
                   
               
               
                 1094 
                 CCCCACCUCCCCGGCCACCCAAA 
                 387 
                   
                 PTPN1:1114L21 sense siNA (1096C) stab19 
               
               
                   
               
               
                 1386 
                 UUACUGGAAGCCCUUCCUGGUCA 
                 388 
                   
                 PTPN1:1406L21 sense siNA (1388C) stab19 
               
               
                   
               
               
                 3035 
                 AGGUGUGGAUAAGGCUUAGGUGC 
                 389 
                   
                 PTPN1:3055L21 sense siNA (3037C) stab19 
               
               
                   
               
               
                  240 
                 UAUCCGACAUGAAGCCAGUGACU 
                 371 
                   
                 PTPN1:260L21 sense siNA (242C) stab22 
               
               
                   
               
               
                  282 
                 GCUUCCUAAGAACAAAAACCGAA 
                 372 
                   
                 PTPN1:302L21 sense siNA (284C) stab22 
               
               
                   
               
               
                  356 
                 AAGAAGAUAAUGACUAUAUCAAC 
                 373 
                   
                 PTPN1:376L21 sense siNA (358C) stab22 
               
               
                   
               
               
                  357 
                 AGAAGAUAAUGACUAUAUCAACG 
                 374 
                   
                 PTPN1:377L21 sense siNA (359C) stab22 
               
               
                   
               
               
                  458 
                 UUUGGGAGAUGGUGUGGGAGCAG 
                 375 
                   
                 PTPN1:478L21 sense siNA (460C) stab22 
               
               
                   
               
               
                  460 
                 UGGGAGAUGGUGUGGGAGCAGAA 
                 376 
                   
                 PTPN1:480L21 sense siNA (462C) stab22 
               
               
                   
               
               
                  708 
                 AUGGCCUGACUUUGGAGUCCCUG 
                 377 
                   
                 PTPN1:728L21 sense siNA (710C) stab22 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                   
                 PTPN1:779L21 sense siNA (761C) stab22 
               
               
                   
               
               
                  764 
                 AAGUCCGAGAGUCAGGGUCACUC 
                 379 
                   
                 PTPN1:784L21 sense siNA (766C) stab22 
               
               
                   
               
               
                  872 
                 UGCUGAUGGACAAGAGGAAAGAC 
                 380 
                   
                 PTPN1:892L21 sense siNA (874C) stab22 
               
               
                   
               
               
                  874 
                 CUGAUGGACAAGAGGAAAGACCC 
                 381 
                   
                 PTPN1:894L21 sense siNA (876C) stab22 
               
               
                   
               
               
                  875 
                 UGAUGGACAAGAGGAAAGACCCU 
                 382 
                   
                 PTPN1:895L21 sense siNA (877C) stab22 
               
               
                   
               
               
                  876 
                 GAUGGACAAGAGGAAAGACCCUU 
                 383 
                   
                 PTPN1:896L21 sense siNA (878C) stab22 
               
               
                   
               
               
                  973 
                 CUGCGCUUCUCCUACCUGGCUGU 
                 384 
                   
                 PTPN1:993L21 sense siNA (975C) stab22 
               
               
                   
               
               
                 1033 
                 GUGCAGGAUCAGUGGAAGGAGCU 
                 385 
                   
                 PTPN1:1053L21 sense siNA (1035C) stab22 
               
               
                   
               
               
                 1035 
                 GCAGGAUCAGUGGAAGGAGCUUU 
                 386 
                   
                 PTPN1:1055L21 sense siNA (1037C) stab22 
               
               
                   
               
               
                 1094 
                 CCCCACCUCCCCGGCCACCCAAA 
                 387 
                   
                 PTPN1:1114L21 sense siNA (1096C) stab22 
               
               
                   
               
               
                 1386 
                 UUACUGGAAGCCCUUCCUGGUCA 
                 388 
                   
                 PTPN1:1406L21 sense siNA (1388C) stab22 
               
               
                   
               
               
                 3035 
                 AGGUGUGGAUAAGGCUUAGGUGC 
                 389 
                   
                 PTPN1:3055L21 sense siNA (3037C) stab22 
               
               
                   
               
               
                  282 
                 GCUUCCUAAGAACAAAAACCGAA 
                 372 
                 39315 
                 PTPN1:302L21 sense siNA (284C) stab25 
               
               
                   
               
               
                  356 
                 AAGAAGAUAAUGACUAUAUCAAC 
                 373 
                 39316 
                 PTPN1:376L21 sense siNA (358C) stab25 
               
               
                   
               
               
                  357 
                 AGAAGAUAAUGACUAUAUCAACG 
                 374 
                 39317 
                 PTPN1:377L21 sense siNA (359C) stab25 
               
               
                   
               
               
                  458 
                 UUUGGGAGAUGGUGUGGGAGCAG 
                 375 
                 39318 
                 PTPN1:478L21 sense siNA (460C) stab25 
               
               
                   
               
               
                  460 
                 UGGGAGAUGGUGUGGGAGCAGAA 
                 376 
                 39319 
                 PTPN1:480L21 sense siNA (462C) stab25 
               
               
                   
               
               
                  708 
                 AUGGCCUGACUUUGGAGUCCCUG 
                 377 
                 39320 
                 PTPN1:728L21 sense siNA (710C) stab25 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                 39321 
                 PTPN1:779L21 sense siNA (761C) stab25 
               
               
                   
               
               
                  872 
                 UGCUGAUGGACAAGAGGAAAGAC 
                 380 
                 39322 
                 PTPN1:892L21 sense siNA (874C) stab25 
               
               
                   
               
               
                  874 
                 CUGAUGGACAAGAGGAAAGACCC 
                 381 
                 39323 
                 PTPN1:894L21 sense siNA (876C) stab25 
               
               
                   
               
               
                  875 
                 UGAUGGACAAGAGGAAAGACCCU 
                 382 
                 39324 
                 PTPN1:895L21 sense siNA (877C) stab25 
               
               
                   
               
               
                  876 
                 GAUGGACAAGAGGAAAGACCCUU 
                 383 
                 39325 
                 PTPN1:896L21 sense siNA (878C) stab25 
               
               
                   
               
               
                  973 
                 CUGCGCUUCUCCUACCUGGCUGU 
                 384 
                 39326 
                 PTPN1:993L21 sense siNA (975C) stab25 
               
               
                   
               
               
                 1033 
                 GUGCAGGAUCAGUGGAAGGAGCU 
                 385 
                 39327 
                 PTPN1:1053L21 sense siNA (1035C) stab25 
               
               
                   
               
               
                 1035 
                 GCAGGAUCAGUGGAAGGAGCUUU 
                 386 
                 39328 
                 PTPN1:1055L21 sense siNA (1037C) stab25 
               
               
                   
               
               
                 1094 
                 CCCCACCUCCCCGGCCACCCAAA 
                 387 
                 39329 
                 PTPN1:1114L21 sense siNA (1096C) stab25 
               
               
                   
               
               
                 1386 
                 UUACUGGAAGCCCUUCCUGGUCA 
                 388 
                 39330 
                 PTPN1:1406L21 sense siNA (1388C) stab25 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                 39756 
                 PTPN1:761U21 sense siNA stab04 + chol TEG 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                 39757 
                 PTPN1:761U21 sense siNA stab07 + chol TEG 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                 39798 
                 PTPN1:761U21 sense siNA stab04 2′OCF3-C 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                 39799 
                 PTPN1:761U21 sense siNA stab07 2′OCF3-C 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                 39800 
                 PTPN1:779L21 sense siNA (761C) stab08 
               
               
                   
                   
                   
                   
                 2′OCF3-C 
               
               
                   
               
               
                  759 
                 UUUCAAAGUCCGAGAGUCAGGGU 
                 378 
                 39801 
                 PTPN1:779L21 sense siNA (761C) stab08 
               
               
                   
                   
                   
                   
                 2′OCF3-C @ 5′ 
               
               
                   
               
               
                  283 
                 AGCUUCCUAAGAACAAAAACCGA 
                 390 
                 40021 
                 PTPN1:283U21 sense siNA stab04 
               
               
                   
               
               
                  461 
                 UUGGGAGAUGGUGUGGGAGCAGA 
                 391 
                 40022 
                 PTPN1:461U21 sense siNA stab04 
               
               
                   
               
               
                  463 
                 GGGAGAUGGUGUGGGAGCAGAAA 
                 392 
                 40023 
                 PTPN1:463U21 sense siNA stab04 
               
               
                   
               
               
                  464 
                 GGAGAUGGUGUGGGAGCAGAAAA 
                 393 
                 40024 
                 PTPN1:464U21 sense siNA stab04 
               
               
                   
               
               
                  703 
                 AUACCACAUGGCCUGACUUUGGA 
                 394 
                 40025 
                 PTPN1:703U21 sense siNA stab04 
               
               
                   
               
               
                  704 
                 UACCACAUGGCCUGACUUUGGAG 
                 395 
                 40026 
                 PTPN1:704U21 sense siNA stab04 
               
               
                   
               
               
                  705 
                 ACCACAUGGCCUGACUUUGGAGU 
                 396 
                 40027 
                 PTPN1:705U21 sense siNA stab04 
               
               
                   
               
               
                  706 
                 CCACAUGGCCUGACUUUGGAGUC 
                 397 
                 40028 
                 PTPN1:706U21 sense siNA stab04 
               
               
                   
               
               
                  707 
                 CACAUGGCCUGACUUUGGAGUCC 
                 398 
                 40029 
                 PTPN1:707U21 sense siNA stab04 
               
               
                   
               
               
                  708 
                 ACAUGGCCUGACUUUGGAGUCCC 
                 399 
                 40030 
                 PTPN1:708U21 sense siNA stab04 
               
               
                   
               
               
                  709 
                 CAUGGCCUGACUUUGGAGUCCCU 
                 400 
                 40031 
                 PTPN1:709U21 sense siNA stab04 
               
               
                   
               
               
                  757 
                 UUCUUUUCAAAGUCCGAGAGUCA 
                 401 
                 40032 
                 PTPN1:757U21 sense siNA stab04 
               
               
                   
               
               
                  758 
                 UCUUUUCAAAGUCCGAGAGUCAG 
                 402 
                 40033 
                 PTPN1:758U21 sense siNA stab04 
               
               
                   
               
               
                  759 
                 CUUUUCAAAGUCCGAGAGUCAGG 
                 403 
                 40034 
                 PTPN1:759U21 sense siNA stab04 
               
               
                   
               
               
                  760 
                 UUUUCAAAGUCCGAGAGUCAGGG 
                 404 
                 40035 
                 PTPN1:760U21 sense siNA stab04 
               
               
                   
               
               
                  875 
                 GCUGAUGGACAAGAGGAAAGACC 
                 405 
                 40036 
                 PTPN1:875U21 sense siNA stab04 
               
               
                   
               
               
                  974 
                 GCUGCGCUUCUCCUACCUGGCUG 
                 406 
                 40037 
                 PTPN1:974U21 sense siNA stab04 
               
               
                   
               
               
                  976 
                 UGCGCUUCUCCUACCUGGCUGUG 
                 407 
                 40038 
                 PTPN1:976U21 sense siNA stab04 
               
               
                   
               
               
                  977 
                 GCGCUUCUCCUACCUGGCUGUGA 
                 408 
                 40039 
                 PTPN1:977U21 sense siNA stab04 
               
               
                   
               
               
                 1033 
                 CCGUGCAGGAUCAGUGGAAGGAG 
                 409 
                 40040 
                 PTPN1:1033U21 sense siNA stab04 
               
               
                   
               
               
                 1034 
                 CGUGCAGGAUCAGUGGAAGGAGC 
                 410 
                 40041 
                 PTPN1:1034U21 sense siNA stab04 
               
               
                   
               
               
                 1036 
                 UGCAGGAUCAGUGGAAGGAGCUU 
                 411 
                 40042 
                 PTPN1:1036U21 sense siNA stab04 
               
               
                   
               
               
                 1389 
                 UACUGGAAGCCCUUCCUGGUCAA 
                 412 
                 40043 
                 PTPN1:1389U21 sense siNA stab04 
               
               
                   
               
               
                 1390 
                 ACUGGAAGCCCUUCCUGGUCAAC 
                 413 
                 40044 
                 PTPN1:1390U21 sense siNA stab04 
               
               
                   
               
               
                 1391 
                 CUGGAAGCCCUUCCUGGUCAACA 
                 414 
                 40045 
                 PTPN1:1391U21 sense siNA stab04 
               
               
                   
               
               
                  283 
                 AGCUUCCUAAGAACAAAAACCGA 
                 390 
                 40046 
                 PTPN1:283U21 sense siNA stab07 
               
               
                   
               
               
                  461 
                 UUGGGAGAUGGUGUGGGAGCAGA 
                 391 
                 40047 
                 PTPN1:461U21 sense siNA stab07 
               
               
                   
               
               
                  463 
                 GGGAGAUGGUGUGGGAGCAGAAA 
                 392 
                 40048 
                 PTPN1:463U21 sense siNA stab07 
               
               
                   
               
               
                  464 
                 GGAGAUGGUGUGGGAGCAGAAAA 
                 393 
                 40049 
                 PTPN1:464U21 sense siNA stab07 
               
               
                   
               
               
                  703 
                 AUACCACAUGGCCUGACUUUGGA 
                 394 
                 40050 
                 PTPN1:703U21 sense siNA stab07 
               
               
                   
               
               
                  704 
                 UACCACAUGGCCUGACUUUGGAG 
                 395 
                 40051 
                 PTPN1:704U21 sense siNA stab07 
               
               
                   
               
               
                  705 
                 ACCACAUGGCCUGACUUUGGAGU 
                 396 
                 40052 
                 PTPN1:705U21 sense siNA stab07 
               
               
                   
               
               
                  706 
                 CCACAUGGCCUGACUUUGGAGUC 
                 397 
                 40053 
                 PTPN1:706U21 sense siNA stab07 
               
               
                   
               
               
                  707 
                 CACAUGGCCUGACUUUGGAGUCC 
                 398 
                 40054 
                 PTPN1:707U21 sense siNA stab07 
               
               
                   
               
               
                  708 
                 ACAUGGCCUGACUUUGGAGUCCC 
                 399 
                 40055 
                 PTPN1:708U21 sense siNA stab07 
               
               
                   
               
               
                  709 
                 CAUGGCCUGACUUUGGAGUCCCU 
                 400 
                 40056 
                 PTPN1:709U21 sense siNA stab07 
               
               
                   
               
               
                  757 
                 UUCUUUUCAAAGUCCGAGAGUCA 
                 401 
                 40057 
                 PTPN1:757U21 sense siNA stab07 
               
               
                   
               
               
                  758 
                 UCUUUUCAAAGUCCGAGAGUCAG 
                 402 
                 40058 
                 PTPN1:758U21 sense siNA stab07 
               
               
                   
               
               
                  759 
                 CUUUUCAAAGUCCGAGAGUCAGG 
                 403 
                 40059 
                 PTPN1:759U21 sense siNA stab07 
               
               
                   
               
               
                  760 
                 UUUUCAAAGUCCGAGAGUCAGGG 
                 404 
                 40060 
                 PTPN1:760U21 sense siNA stab07 
               
               
                   
               
               
                  875 
                 GCUGAUGGACAAGAGGAAAGACC 
                 405 
                 40061 
                 PTPN1:875U21 sense siNA stab07 
               
               
                   
               
               
                  974 
                 GCUGCGCUUCUCCUACCUGGCUG 
                 406 
                 40062 
                 PTPN1:974U21 sense siNA stab07 
               
               
                   
               
               
                  976 
                 UGCGCUUCUCCUACCUGGCUGUG 
                 407 
                 40063 
                 PTPN1:976U21 sense siNA stab07 
               
               
                   
               
               
                  977 
                 GCGCUUCUCCUACCUGGCUGUGA 
                 408 
                 40064 
                 PTPN1:977U21 sense siNA stab07 
               
               
                   
               
               
                 1033 
                 CCGUGCAGGAUCAGUGGAAGGAG 
                 409 
                 40065 
                 PTPN1:1033U21 sense siNA stab07 
               
               
                   
               
               
                 1034 
                 CGUGCAGGAUCAGUGGAAGGAGC 
                 410 
                 40066 
                 PTPN1:1034U21 sense siNA stab07 
               
               
                   
               
               
                 1036 
                 UGCAGGAUCAGUGGAAGGAGCUU 
                 411 
                 40067 
                 PTPN1:1036U21 sense siNA stab07 
               
               
                   
               
               
                 1389 
                 UACUGGAAGCCCUUCCUGGUCAA 
                 412 
                 40068 
                 PTPN1:1389U21 sense siNA stab07 
               
               
                   
               
               
                 1390 
                 ACUGGAAGCCCUUCCUGGUCAAC 
                 413 
                 40069 
                 PTPN1:1390U21 sense siNA stab07 
               
               
                   
               
               
                 1391 
                 CUGGAAGCCCUUCCUGGUCAACA 
                 414 
                 40070 
                 PTPN1:1391U21 sense siNA stab07 
               
               
                   
               
               
                  283 
                 AGCUUCCUAAGAACAAAAACCGA 
                 390 
                 40071 
                 PTPN1:301L21 sense siNA (283C) stab25 
               
               
                   
               
               
                  461 
                 UUGGGAGAUGGUGUGGGAGCAGA 
                 391 
                 40072 
                 PTPN1:479L21 sense siNA (461C) stab25 
               
               
                   
               
               
                  463 
                 GGGAGAUGGUGUGGGAGCAGAAA 
                 392 
                 40073 
                 PTPN1:481L21 sense siNA (463C) stab25 
               
               
                   
               
               
                  464 
                 GGAGAUGGUGUGGGAGCAGAAAA 
                 393 
                 40074 
                 PTPN1:482L21 sense siNA (464C) stab25 
               
               
                   
               
               
                  703 
                 AUACCACAUGGCCUGACUUUGGA 
                 394 
                 40075 
                 PTPN1:721L21 sense siNA (703C) stab25 
               
               
                   
               
               
                  704 
                 UACCACAUGGCCUGACUUUGGAG 
                 395 
                 40076 
                 PTPN1:722L21 sense siNA (704C) stab25 
               
               
                   
               
               
                  705 
                 ACCACAUGGCCUGACUUUGGAGU 
                 396 
                 40077 
                 PTPN1:723L21 sense siNA (705C) stab25 
               
               
                   
               
               
                  706 
                 CCACAUGGCCUGACUUUGGAGUC 
                 397 
                 40078 
                 PTPN1:724L21 sense siNA (706C) stab25 
               
               
                   
               
               
                  707 
                 CACAUGGCCUGACUUUGGAGUCC 
                 398 
                 40079 
                 PTPN1:725L21 sense siNA (707C) stab25 
               
               
                   
               
               
                  708 
                 ACAUGGCCUGACUUUGGAGUCCC 
                 399 
                 40080 
                 PTPN1:726L21 sense siNA (708C) stab25 
               
               
                   
               
               
                  709 
                 CAUGGCCUGACUUUGGAGUCCCU 
                 400 
                 40081 
                 PTPN1:727L21 sense siNA (709C) stab25 
               
               
                   
               
               
                  757 
                 UUCUUUUCAAAGUCCGAGAGUCA 
                 401 
                 40082 
                 PTPN1:775L21 sense siNA (757C) stab25 
               
               
                   
               
               
                  758 
                 UCUUUUCAAAGUCCGAGAGUCAG 
                 402 
                 40083 
                 PTPN1:776L21 sense siNA (758C) stab25 
               
               
                   
               
               
                  759 
                 CUUUUCAAAGUCCGAGAGUCAGG 
                 403 
                 40084 
                 PTPN1:777L21 sense siNA (759C) stab25 
               
               
                   
               
               
                  760 
                 UUUUCAAAGUCCGAGAGUCAGGG 
                 404 
                 40085 
                 PTPN1:778L21 sense siNA (760C) stab25 
               
               
                   
               
               
                  875 
                 GCUGAUGGACAAGAGGAAAGACC 
                 405 
                 40086 
                 PTPN1:893L21 sense siNA (875C) stab25 
               
               
                   
               
               
                  974 
                 GCUGCGCUUCUCCUACCUGGCUG 
                 406 
                 40087 
                 PTPN1:992L21 sense siNA (974C) stab25 
               
               
                   
               
               
                  976 
                 UGCGCUUCUCCUACCUGGCUGUG 
                 407 
                 40088 
                 PTPN1:994L21 sense siNA (976C) stab25 
               
               
                   
               
               
                  977 
                 GCGCUUCUCCUACCUGGCUGUGA 
                 408 
                 40089 
                 PTPN1:995L21 sense siNA (977C) stab25 
               
               
                   
               
               
                 1033 
                 CCGUGCAGGAUCAGUGGAAGGAG 
                 409 
                 40090 
                 PTPN1:1051L21 sense siNA (1033C) stab25 
               
               
                   
               
               
                 1034 
                 CGUGCAGGAUCAGUGGAAGGAGC 
                 410 
                 40091 
                 PTPN1:1052L21 sense siNA (1034C) stab25 
               
               
                   
               
               
                 1036 
                 UGCAGGAUCAGUGGAAGGAGCUU 
                 411 
                 40092 
                 PTPN1:1054L21 sense siNA (1036C) stab25 
               
               
                   
               
               
                 1389 
                 UACUGGAAGCCCUUCCUGGUCAA 
                 412 
                 40093 
                 PTPN1:1407L21 sense siNA (1389C) stab25 
               
               
                   
               
               
                 1390 
                 ACUGGAAGCCCUUCCUGGUCAAC 
                 413 
                 40094 
                 PTPN1:1408L21 sense siNA (1390C) stab25 
               
               
                   
               
               
                 1391 
                 CUGGAAGCCCUUCCUGGUCAACA 
                 414 
                 40095 
                 PTPN1:1409L21 sense siNA (1391C) stab25 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                   
                 Target Pos 
                 Sequence 
                 Seq ID 
               
               
                   
                   
               
               
                   
                  240 
                 UCCGACAUGAAGCCAGUGATT 
                 415 
               
               
                   
                   
               
               
                   
                  282 
                 UUCCUAAGAACAAAAACCGTT 
                 416 
               
               
                   
                   
               
               
                   
                  356 
                 GAAGAUAAUGACUAUAUCATT 
                 417 
               
               
                   
                   
               
               
                   
                  357 
                 AAGAUAAUGACUAUAUCAATT 
                 418 
               
               
                   
                   
               
               
                   
                  458 
                 UGGGAGAUGGUGUGGGAGCTT 
                 419 
               
               
                   
                   
               
               
                   
                  460 
                 GGAGAUGGUGUGGGAGCAGTT 
                 420 
               
               
                   
                   
               
               
                   
                  708 
                 GGCCUGACUUUGGAGUCCCTT 
                 421 
               
               
                   
                   
               
               
                   
                  759 
                 UCAAAGUCCGAGAGUCAGGTT 
                 422 
               
               
                   
                   
               
               
                   
                  764 
                 GUCCGAGAGUCAGGGUCACTT 
                 423 
               
               
                   
                   
               
               
                   
                  872 
                 CUGAUGGACAAGAGGAAAGTT 
                 424 
               
               
                   
                   
               
               
                   
                  874 
                 GAUGGACAAGAGGAAAGACTT 
                 425 
               
               
                   
                   
               
               
                   
                  875 
                 AUGGACAAGAGGAAAGACCTT 
                 426 
               
               
                   
                   
               
               
                   
                  876 
                 UGGACAAGAGGAAAGACCCTT 
                 427 
               
               
                   
                   
               
               
                   
                  973 
                 GCGCUUCUCCUACCUGGCUTT 
                 428 
               
               
                   
                   
               
               
                   
                 1033 
                 GCAGGAUCAGUGGAAGGAGTT 
                 429 
               
               
                   
                   
               
               
                   
                 1035 
                 AGGAUCAGUGGAAGGAGCUTT 
                 430 
               
               
                   
                   
               
               
                   
                 1094 
                 CCACCUCCCCGGCCACCCATT 
                 431 
               
               
                   
                   
               
               
                   
                 1386 
                 ACUGGAAGCCCUUCCUGGUTT 
                 432 
               
               
                   
                   
               
               
                   
                 3035 
                 GUGUGGAUAAGGCUUAGGUTT 
                 433 
               
               
                   
                   
               
               
                   
                  240 
                 UCACUGGCUUCAUGUCGGATT 
                 434 
               
               
                   
                   
               
               
                   
                  282 
                 CGGUUUUUGUUCUUAGGAATT 
                 435 
               
               
                   
                   
               
               
                   
                  356 
                 UGAUAUAGUCAUUAUCUUCTT 
                 436 
               
               
                   
                   
               
               
                   
                  357 
                 UUGAUAUAGUCAUUAUCUUTT 
                 437 
               
               
                   
                   
               
               
                   
                  458 
                 GCUCCCACACCAUCUCCCATT 
                 438 
               
               
                   
                   
               
               
                   
                  460 
                 CUGCUCCCACACCAUCUCCTT 
                 439 
               
               
                   
                   
               
               
                   
                  708 
                 GGGACUCCAAAGUCAGGCCTT 
                 440 
               
               
                   
                   
               
               
                   
                  759 
                 CCUGACUCUCGGACUUUGATT 
                 441 
               
               
                   
                   
               
               
                   
                  764 
                 GUGACCCUGACUCUCGGACTT 
                 442 
               
               
                   
                   
               
               
                   
                  872 
                 CUUUCCUCUUGUCCAUCAGTT 
                 443 
               
               
                   
                   
               
               
                   
                  874 
                 GUCUUUCCUCUUGUCCAUCTT 
                 444 
               
               
                   
                   
               
               
                   
                  875 
                 GGUCUUUCCUCUUGUCCAUTT 
                 445 
               
               
                   
                   
               
               
                   
                  876 
                 GGGUCUUUCCUCUUGUCCATT 
                 446 
               
               
                   
                   
               
               
                   
                  973 
                 AGCCAGGUAGGAGAAGCGCTT 
                 447 
               
               
                   
                   
               
               
                   
                 1033 
                 CUCCUUCCACUGAUCCUGCTT 
                 448 
               
               
                   
                   
               
               
                   
                 1035 
                 AGCUCCUUCCACUGAUCCUTT 
                 449 
               
               
                   
                   
               
               
                   
                 1094 
                 UGGGUGGCCGGGGAGGUGGTT 
                 450 
               
               
                   
                   
               
               
                   
                 1386 
                 ACCAGGAAGGGCUUCCAGUTT 
                 451 
               
               
                   
                   
               
               
                   
                 3035 
                 ACCUAAGCCUUAUCCACACTT 
                 452 
               
               
                   
                   
               
               
                   
                  240 
                 B uccGAcAuGAAGccAGuGATT B 
                 453 
               
               
                   
                   
               
               
                   
                  282 
                 B uuccuAAGAAcAAAAAccGTT B 
                 454 
               
               
                   
                   
               
               
                   
                  356 
                 B GAAGAuAAuGAcuAuAucATT B 
                 455 
               
               
                   
                   
               
               
                   
                  357 
                 B AAGAuAAuGAcuAuAucAATT B 
                 456 
               
               
                   
                   
               
               
                   
                  458 
                 B uGGGAGAuGGuGuGGGAGcTT B 
                 457 
               
               
                   
                   
               
               
                   
                  460 
                 B GGAGAuGGuGuGGGAGcAGTT B 
                 458 
               
               
                   
                   
               
               
                   
                  708 
                 B GGccuGAcuuuGGAGucccTT B 
                 459 
               
               
                   
                   
               
               
                   
                  759 
                 B ucAAAGuccGAGAGucAGGTT B 
                 460 
               
               
                   
                   
               
               
                   
                  764 
                 B GuccGAGAGucAGGGucAcTT B 
                 461 
               
               
                   
                   
               
               
                   
                  872 
                 B cuGAuGGAcAAGAGGAAAGTT B 
                 462 
               
               
                   
                   
               
               
                   
                  874 
                 B GAuGGAcAAGAGGAAAGAcTT B 
                 463 
               
               
                   
                   
               
               
                   
                  875 
                 B AuGGAcAAGAGGAAAGAccTT B 
                 464 
               
               
                   
                   
               
               
                   
                  876 
                 B uGGAcAAGAGGAAAGAcccTT B 
                 465 
               
               
                   
                   
               
               
                   
                  973 
                 B GcGcuucuccuAccuGGcuTT B 
                 466 
               
               
                   
                   
               
               
                   
                 1033 
                 B GcAGGAucAGuGGAAGGAGTT B 
                 467 
               
               
                   
                   
               
               
                   
                 1035 
                 B AGGAucAGuGGAAGGAGcuTT B 
                 468 
               
               
                   
                   
               
               
                   
                 1094 
                 B ccAccuccccGGccAcccATT B 
                 469 
               
               
                   
                   
               
               
                   
                 1386 
                 B AcuGGAAGcccuuccuGGuTT B 
                 470 
               
               
                   
                   
               
               
                   
                 3035 
                 B GuGuGGAuAAGGcuuAGGuTT B 
                 471 
               
               
                   
                   
               
               
                   
                  240 
                 ucAcuGGcuucAuGucGGATsT 
                 472 
               
               
                   
                   
               
               
                   
                  282 
                 cGGuuuuuGuucuuAGGAATsT 
                 473 
               
               
                   
                   
               
               
                   
                  356 
                 uGAuAuAGucAuuAucuucTsT 
                 474 
               
               
                   
                   
               
               
                   
                  357 
                 uuGAuAuAGucAuuAucuuTsT 
                 475 
               
               
                   
                   
               
               
                   
                  458 
                 GcucccAcAccAucucccATsT 
                 476 
               
               
                   
                   
               
               
                   
                  460 
                 cuGcucccAcAccAucuccTsT 
                 477 
               
               
                   
                   
               
               
                   
                  708 
                 GGGAcuccAAAGucAGGccTsT 
                 478 
               
               
                   
                   
               
               
                   
                  759 
                 ccuGAcucucGGAcuuuGATsT 
                 479 
               
               
                   
                   
               
               
                   
                  764 
                 GuGAcccuGAcucucGGAcTsT 
                 480 
               
               
                   
                   
               
               
                   
                  872 
                 cuuuccucuuGuccAucAGTsT 
                 481 
               
               
                   
                   
               
               
                   
                  874 
                 GucuuuccucuuGuccAucTsT 
                 482 
               
               
                   
                   
               
               
                   
                  875 
                 GGucuuuccucuuGuccAuTsT 
                 483 
               
               
                   
                   
               
               
                   
                  876 
                 GGGucuuuccucuuGuccATsT 
                 484 
               
               
                   
                   
               
               
                   
                  973 
                 AGccAGGuAGGAGAAGcGcTsT 
                 485 
               
               
                   
                   
               
               
                   
                 1033 
                 cuccuuccAcuGAuccuGcTsT 
                 486 
               
               
                   
                   
               
               
                   
                 1035 
                 AGcuccuuccAcuGAuccuTsT 
                 487 
               
               
                   
                   
               
               
                   
                 1094 
                 uGGGuGGccGGGGAGGuGGTsT 
                 488 
               
               
                   
                   
               
               
                   
                 1386 
                 AccAGGAAGGGcuuccAGuTsT 
                 489 
               
               
                   
                   
               
               
                   
                 3035 
                 AccuAAGccuuAuccAcAcTsT 
                 490 
               
               
                   
                   
               
               
                   
                  240 
                 B ucc GA c A u GAAG cc AG u GA TT B 
                 491 
               
               
                   
                   
               
               
                   
                  282 
                 B uuccu AAGAA c AAAAA cc G TT B 
                 492 
               
               
                   
                   
               
               
                   
                  356 
                 B  GAAGA u AA u GA cu A u A uc A TT B 
                 493 
               
               
                   
                   
               
               
                   
                  357 
                 B  AAGA u AA u GA cu A u A uc AA TT B 
                 494 
               
               
                   
                   
               
               
                   
                  458 
                 B u GGGAGA u GG u G u GGGAG cTT B 
                 495 
               
               
                   
                   
               
               
                   
                  460 
                 B  GGAGA u GG u G u GGGAG c AG TT B 
                 496 
               
               
                   
                   
               
               
                   
                  708 
                 B  GG ccu GA cuuu GGAG ucccTT B 
                 497 
               
               
                   
                   
               
               
                   
                  759 
                 B uc AAAG ucc GAGAG uc AGG TT B 
                 498 
               
               
                   
                   
               
               
                   
                  764 
                 B  G ucc GAGAG uc AGGG uc A cTT B 
                 499 
               
               
                   
                   
               
               
                   
                  872 
                 B cu GA u GGA c AAGAGGAAAG TT B 
                 500 
               
               
                   
                   
               
               
                   
                  874 
                 B  GA u GGA c AAGAGGAAAGA cTT B 
                 501 
               
               
                   
                   
               
               
                   
                  875 
                 B  A u GGA c AAGAGGAAAGA ccTT B 
                 502 
               
               
                   
                   
               
               
                   
                  876 
                 B u GGA c AAGAGGAAAGA cccTT B 
                 503 
               
               
                   
                   
               
               
                   
                  973 
                 B  G c G cuucuccu A ccu GG cuTT B 
                 504 
               
               
                   
                   
               
               
                   
                 1033 
                 B  G c AGGA uc AG u GGAAGGAG TT B 
                 505 
               
               
                   
                   
               
               
                   
                 1035 
                 B  AGGA uc AG u GGAAGGAG cuTT B 
                 506 
               
               
                   
                   
               
               
                   
                 1094 
                 B cc Accucccc   GG cc A ccc A TT B 
                 507 
               
               
                   
                   
               
               
                   
                 1386 
                 B  A cu GGAAG cccuuccu GG uTT B 
                 508 
               
               
                   
                   
               
               
                   
                 3035 
                 B  G u G u GGA u AAGG cuu AGG uTT B 
                 509 
               
               
                   
                   
               
               
                   
                  240 
                 uc A cu GG cuuc A u G uc GGA TsT 
                 510 
               
               
                   
                   
               
               
                   
                  282 
                 c GG uuuuu G uucuu AGGAA TsT 
                 511 
               
               
                   
                   
               
               
                   
                  356 
                 u GA u A u AG uc A uu A ucuucTsT 
                 512 
               
               
                   
                   
               
               
                   
                  357 
                 uu GA u A u AG uc A uu A ucuuTsT 
                 513 
               
               
                   
                   
               
               
                   
                  458 
                   G cuccc A c A cc A ucuccc A TsT 
                 514 
               
               
                   
                   
               
               
                   
                  460 
                 cu G cuccc A c A cc A ucuccTsT 
                 515 
               
               
                   
                   
               
               
                   
                  708 
                   GGGA cucc AAAG uc AGG ccTsT 
                 516 
               
               
                   
                   
               
               
                   
                  759 
                 ccu GA cucuc GGA cuuu GA TsT 
                 517 
               
               
                   
                   
               
               
                   
                  764 
                   G u GA cccu GA cucuc GGA cTsT 
                 518 
               
               
                   
                   
               
               
                   
                  872 
                 cuuuccucuu G ucc A uc AG TsT 
                 519 
               
               
                   
                   
               
               
                   
                  874 
                   G ucuuuccucuu G ucc A ucTsT 
                 520 
               
               
                   
                   
               
               
                   
                  875 
                   GG ucuuuccucuu G ucc A uTsT 
                 521 
               
               
                   
                   
               
               
                   
                  876 
                   GGG ucuuuccucuu G ucc A TsT 
                 522 
               
               
                   
                   
               
               
                   
                  973 
                   AG cc AGG u AGGAGAAG c G cTsT 
                 523 
               
               
                   
                   
               
               
                   
                 1033 
                 cuccuucc A cu GA uccu G cTsT 
                 524 
               
               
                   
                   
               
               
                   
                 1035 
                   AG cuccuucc A cu GA uccuTsT 
                 525 
               
               
                   
                   
               
               
                   
                 1094 
                 u GGG u GG cc GGGGAGG u GG TsT 
                 526 
               
               
                   
                   
               
               
                   
                 1386 
                   A cc AGGAAGGG cuucc AG uTsT 
                 527 
               
               
                   
                   
               
               
                   
                 3035 
                   A ccu AAG ccuu A ucc A c A cTsT 
                 528 
               
               
                   
                   
               
               
                   
                  240 
                 B ucc GA c A u GAAG cc AG u GA TT B 
                 529 
               
               
                   
                   
               
               
                   
                  282 
                 B uuccu AAGAA c AAAAA cc G TT B 
                 530 
               
               
                   
                   
               
               
                   
                  356 
                 B  GAAGA u AA u GA cu A u A uc A TT B 
                 531 
               
               
                   
                   
               
               
                   
                  357 
                 B  AAGA u AA u GA cu A u A uc AA TT B 
                 532 
               
               
                   
                   
               
               
                   
                  458 
                 B u GGGAGA u GG u G u GGGAG cTT B 
                 533 
               
               
                   
                   
               
               
                   
                  460 
                 B  GGAGA u GG u G u GGGAG c AG TT B 
                 534 
               
               
                   
                   
               
               
                   
                  708 
                 B  GG ccu GA cuuu GGAG ucccTT B 
                 535 
               
               
                   
                   
               
               
                   
                  759 
                 B uc AAAG ucc GAGAG uc AGG TT B 
                 536 
               
               
                   
                   
               
               
                   
                  764 
                 B  G ucc GAGAG uc AGGG uc A cTT B 
                 537 
               
               
                   
                   
               
               
                   
                  872 
                 B cu GA u GGA c AAGAGGAAAG TT B 
                 538 
               
               
                   
                   
               
               
                   
                  874 
                 B  GA u GGA c AAGAGGAAAGA cTT B 
                 539 
               
               
                   
                   
               
               
                   
                  875 
                 B  A u GGA c AAGAGGAAAGA ccTT B 
                 540 
               
               
                   
                   
               
               
                   
                  876 
                 B u GGA c AAGAGGAAAGA cccTT B 
                 541 
               
               
                   
                   
               
               
                   
                  973 
                 B  G c G cuucuccu A ccu GG cuTT B 
                 542 
               
               
                   
                   
               
               
                   
                 1033 
                 B  G c AGGA uc AG u GGAAGGAG TT B 
                 543 
               
               
                   
                   
               
               
                   
                 1035 
                 B  AGGA uc AG u GGAAGGAG cuTT B 
                 544 
               
               
                   
                   
               
               
                   
                 1094 
                 B cc A ccucccc GG cc A ccc A TT B 
                 545 
               
               
                   
                   
               
               
                   
                 1386 
                 B  A cu GGAAG cccuuccu GG uTT B 
                 546 
               
               
                   
                   
               
               
                   
                 3035 
                 B  G u G u GGA u AAGG cuu AGG uTT B 
                 547 
               
               
                   
                   
               
               
                   
                  240 
                 uc A cu GG cuuc A u G uc GGA TsT 
                 548 
               
               
                   
                   
               
               
                   
                  282 
                 c GG uuuuu G uucuu AGGAA TsT 
                 549 
               
               
                   
                   
               
               
                   
                  356 
                 u GA u A u AG uc A uu A ucuucTsT 
                 550 
               
               
                   
                   
               
               
                   
                  357 
                 uu GA u A u AG uc A uu A ucuuTsT 
                 551 
               
               
                   
                   
               
               
                   
                  458 
                   G cuccc A c A cc A ucuccc A TsT 
                 552 
               
               
                   
                   
               
               
                   
                  460 
                 cu G cuccc A c A cc A ucuccTsT 
                 553 
               
               
                   
                   
               
               
                   
                  708 
                   GGGA cucc AAAG uc AGG ccTsT 
                 554 
               
               
                   
                   
               
               
                   
                  759 
                 ccu GA cucuc GGA cuuu GA TsT 
                 555 
               
               
                   
                   
               
               
                   
                  764 
                   G u GA cccu GA cucuc GGA cTsT 
                 556 
               
               
                   
                   
               
               
                   
                  872 
                 cuuuccucuu G ucc A uc AG TsT 
                 557 
               
               
                   
                   
               
               
                   
                  874 
                   G ucuuuccucuu G ucc A ucTsT 
                 558 
               
               
                   
                   
               
               
                   
                  875 
                   GG ucuuuccucuu G ucc A uTsT 
                 559 
               
               
                   
                   
               
               
                   
                  876 
                   GGG ucuuuccucuu G ucc A TsT 
                 560 
               
               
                   
                   
               
               
                   
                  973 
                   AG cc AGG u AGGAGAAG c G cTsT 
                 561 
               
               
                   
                   
               
               
                   
                 1033 
                 cuccuucc A cu GA uccu G cTsT 
                 562 
               
               
                   
                   
               
               
                   
                 1035 
                   AG cuccuucc A cu GA uccuTsT 
                 563 
               
               
                   
                   
               
               
                   
                 1094 
                 u GGG u GG cc GGGGAGG u GG TsT 
                 564 
               
               
                   
                   
               
               
                   
                 1386 
                   A cc AGGAAGGG cuucc AG uTsT 
                 565 
               
               
                   
                   
               
               
                   
                 3035 
                   A ccu AAG ccuu A ucc A c A cTsT 
                 566 
               
               
                   
                   
               
               
                   
                  240 
                 B UCCGACAUGAAGCCAGUGATT B 
                 567 
               
               
                   
                   
               
               
                   
                  282 
                 B UUCCUAAGAACAAAAACCGTT B 
                 568 
               
               
                   
                   
               
               
                   
                  356 
                 B GAAGAUAAUGACUAUAUCATT B 
                 569 
               
               
                   
                   
               
               
                   
                  357 
                 B AAGAUAAUGACUAUAUCAATT B 
                 570 
               
               
                   
                   
               
               
                   
                  458 
                 B UGGGAGAUGGUGUGGGAGCTT B 
                 571 
               
               
                   
                   
               
               
                   
                  460 
                 B GGAGAUGGUGUGGGAGCAGTT B 
                 572 
               
               
                   
                   
               
               
                   
                  708 
                 B GGCCUGACUUUGGAGUCCCTT B 
                 573 
               
               
                   
                   
               
               
                   
                  759 
                 B UCAAAGUCCGAGAGUCAGGTT B 
                 574 
               
               
                   
                   
               
               
                   
                  764 
                 B GUCCGAGAGUCAGGGUCACTT B 
                 575 
               
               
                   
                   
               
               
                   
                  872 
                 B CUGAUGGACAAGAGGAAAGTT B 
                 576 
               
               
                   
                   
               
               
                   
                  874 
                 B GAUGGACAAGAGGAAAGACTT B 
                 577 
               
               
                   
                   
               
               
                   
                  875 
                 B AUGGACAAGAGGAAAGACCTT B 
                 578 
               
               
                   
                   
               
               
                   
                  876 
                 B UGGACAAGAGGAAAGACCCTT B 
                 579 
               
               
                   
                   
               
               
                   
                  973 
                 B GCGCUUCUCCUACCUGGCUTT B 
                 580 
               
               
                   
                   
               
               
                   
                 1033 
                 B GCAGGAUCAGUGGAAGGAGTT B 
                 581 
               
               
                   
                   
               
               
                   
                 1035 
                 B AGGAUCAGUGGAAGGAGCUTT B 
                 582 
               
               
                   
                   
               
               
                   
                 1094 
                 B CCACCUCCCCGGCCACCCATT B 
                 583 
               
               
                   
                   
               
               
                   
                 1386 
                 B ACUGGAAGCCCUUCCUGGUTT B 
                 584 
               
               
                   
                   
               
               
                   
                 3035 
                 B GUGUGGAUAAGGCUUAGGUTT B 
                 585 
               
               
                   
                   
               
               
                   
                  240 
                 UCACUGGCUUCAUGUCGGATsT 
                 586 
               
               
                   
                   
               
               
                   
                  282 
                 CGGUUUUUGUUCUUAGGAATsT 
                 587 
               
               
                   
                   
               
               
                   
                  356 
                 UGAUAUAGUCAUUAUCUUCTsT 
                 588 
               
               
                   
                   
               
               
                   
                  357 
                 UUGAUAUAGUCAUUAUCUUTsT 
                 589 
               
               
                   
                   
               
               
                   
                  458 
                 GCUCCCACACCAUCUCCCATsT 
                 590 
               
               
                   
                   
               
               
                   
                  460 
                 CUGCUCCCACACCAUCUCCTsT 
                 591 
               
               
                   
                   
               
               
                   
                  708 
                 GGGACUCCAAAGUCAGGCCTsT 
                 592 
               
               
                   
                   
               
               
                   
                  759 
                 CCUGACUCUCGGACUUUGATsT 
                 593 
               
               
                   
                   
               
               
                   
                  764 
                 GUGACCCUGACUCUCGGACTsT 
                 594 
               
               
                   
                   
               
               
                   
                  872 
                 CUUUCCUCUUGUCCAUCAGTsT 
                 595 
               
               
                   
                   
               
               
                   
                  874 
                 GUCUUUCCUCUUGUCCAUCTsT 
                 596 
               
               
                   
                   
               
               
                   
                  875 
                 GGUCUUUCCUCUUGUCCAUTsT 
                 597 
               
               
                   
                   
               
               
                   
                  876 
                 GGGUCUUUCCUCUUGUCCATsT 
                 598 
               
               
                   
                   
               
               
                   
                  973 
                 AGCCAGGUAGGAGAAGCGCTsT 
                 599 
               
               
                   
                   
               
               
                   
                 1033 
                 CUCCUUCCACUGAUCCUGCTsT 
                 600 
               
               
                   
                   
               
               
                   
                 1035 
                 AGCUCCUUCCACUGAUCCUTsT 
                 601 
               
               
                   
                   
               
               
                   
                 1094 
                 UGGGUGGCCGGGGAGGUGGTsT 
                 602 
               
               
                   
                   
               
               
                   
                 1386 
                 ACCAGGAAGGGCUUCCAGUTsT 
                 603 
               
               
                   
                   
               
               
                   
                 3035 
                 ACCUAAGCCUUAUCCACACTsT 
                 604 
               
               
                   
                   
               
               
                   
                  240 
                 uc A cu GG cuuc A u G uc GGA TT B 
                 605 
               
               
                   
                   
               
               
                   
                  282 
                 c GG uuuuu G uucuu AGGAA TT B 
                 606 
               
               
                   
                   
               
               
                   
                  356 
                 u GA u A u AG uc A uu A ucuucTT B 
                 607 
               
               
                   
                   
               
               
                   
                  357 
                 uu GA u A u AG uc A uu A ucuuTT B 
                 608 
               
               
                   
                   
               
               
                   
                  458 
                   G cuccc A c A cc A ucuccc A TT B 
                 609 
               
               
                   
                   
               
               
                   
                  460 
                 cu G cuccc A c A cc A ucuccTT B 
                 610 
               
               
                   
                   
               
               
                   
                  708 
                   GGGA cucc AAAG uc AGG ccTT B 
                 611 
               
               
                   
                   
               
               
                   
                  759 
                 ccu GA cucuc GGA cuuu GA TT B 
                 612 
               
               
                   
                   
               
               
                   
                  764 
                   G u GA cccu GA cucuc GGA cTT B 
                 613 
               
               
                   
                   
               
               
                   
                  872 
                 cuuuccucuu G ucc A uc AG TT B 
                 614 
               
               
                   
                   
               
               
                   
                  874 
                   G ucuuuccucuu G ucc A ucTT B 
                 615 
               
               
                   
                   
               
               
                   
                  875 
                   GG ucuuuccucuu G ucc A uTT B 
                 616 
               
               
                   
                   
               
               
                   
                  876 
                   GGG ucuuuccucuu G ucc A TT B 
                 617 
               
               
                   
                   
               
               
                   
                  973 
                   AG cc AGG u AGGAGAAG c G cTT B 
                 618 
               
               
                   
                   
               
               
                   
                 1033 
                 cuccuucc A cu GA uccu G cTT B 
                 619 
               
               
                   
                   
               
               
                   
                 1035 
                   AG cuccuucc A cu GA uccuTT B 
                 620 
               
               
                   
                   
               
               
                   
                 1094 
                 u GGG u GG cc GGGGAGG u GG TT B 
                 621 
               
               
                   
                   
               
               
                   
                 1386 
                   A cc AGGAAGGG cuucc AG uTT B 
                 622 
               
               
                   
                   
               
               
                   
                 3035 
                   A ccu AAG ccuu A ucc A c A cTT B 
                 623 
               
               
                   
                   
               
               
                   
                  240 
                 UCACUGGCUUCAUGUCGGATT B 
                 624 
               
               
                   
                   
               
               
                   
                  282 
                 CGGUUUUUGUUCUUAGGAATT B 
                 625 
               
               
                   
                   
               
               
                   
                  356 
                 UGAUAUAGUCAUUAUCUUCTT B 
                 626 
               
               
                   
                   
               
               
                   
                  357 
                 UUGAUAUAGUCAUUAUCUUTT B 
                 627 
               
               
                   
                   
               
               
                   
                  458 
                 GCUCCCACACCAUCUCCCATT B 
                 628 
               
               
                   
                   
               
               
                   
                  460 
                 CUGCUCCCACACCAUCUCCTT B 
                 629 
               
               
                   
                   
               
               
                   
                  708 
                 GGGACUCCAAAGUCAGGCCTT B 
                 630 
               
               
                   
                   
               
               
                   
                  759 
                 CCUGACUCUCGGACUUUGATT B 
                 631 
               
               
                   
                   
               
               
                   
                  764 
                 GUGACCCUGACUCUCGGACTT B 
                 632 
               
               
                   
                   
               
               
                   
                  872 
                 CUUUCCUCUUGUCCAUCAGTT B 
                 633 
               
               
                   
                   
               
               
                   
                  874 
                 GUCUUUCCUCUUGUCCAUCTT B 
                 634 
               
               
                   
                   
               
               
                   
                  875 
                 GGUCUUUCCUCUUGUCCAUTT B 
                 635 
               
               
                   
                   
               
               
                   
                  876 
                 GGGUCUUUCCUCUUGUCCATT B 
                 636 
               
               
                   
                   
               
               
                   
                  973 
                 AGCCAGGUAGGAGAAGCGCTT B 
                 637 
               
               
                   
                   
               
               
                   
                 1033 
                 CUCCUUCCACUGAUCCUGCTT B 
                 638 
               
               
                   
                   
               
               
                   
                 1035 
                 AGCUCCUUCCACUGAUCCUTT B 
                 639 
               
               
                   
                   
               
               
                   
                 1094 
                 UGGGUGGCCGGGGAGGUGGTT B 
                 640 
               
               
                   
                   
               
               
                   
                 1386 
                 ACCAGGAAGGGCUUCCAGUTT B 
                 641 
               
               
                   
                   
               
               
                   
                 3035 
                 ACCUAAGCCUUAUCCACACTT B 
                 642 
               
               
                   
                   
               
               
                   
                  282 
                 CGGuuuuu G uucuu AGGAA TsT 
                 643 
               
               
                   
                   
               
               
                   
                  356 
                 UGAu A u AG uc A uu A ucuucTsT 
                 644 
               
               
                   
                   
               
               
                   
                  357 
                 UUGAu A u AG uc A uu A ucuuTsT 
                 645 
               
               
                   
                   
               
               
                   
                  458 
                 GCUccc A c A cc A ucuccc A TsT 
                 646 
               
               
                   
                   
               
               
                   
                  460 
                 CUGcuccc A c A cc A ucuccTsT 
                 647 
               
               
                   
                   
               
               
                   
                  708 
                 GGG A cucc AAAG uc AGG ccTsT 
                 648 
               
               
                   
                   
               
               
                   
                  759 
                 CCU GA cucuc GGA cuuu GA TsT 
                 649 
               
               
                   
                   
               
               
                   
                  872 
                 CUUuccucuu G ucc A uc AG TsT 
                 650 
               
               
                   
                   
               
               
                   
                  874 
                 GUCuuuccucuu G ucc A ucTsT 
                 651 
               
               
                   
                   
               
               
                   
                  875 
                 GGUcuuuccucuu G ucc A uTsT 
                 652 
               
               
                   
                   
               
               
                   
                  876 
                 GGGucuuuccucuu G ucc A TsT 
                 653 
               
               
                   
                   
               
               
                   
                  973 
                 AGCc AGG u AGGAGAAG c G cTsT 
                 654 
               
               
                   
                   
               
               
                   
                 1033 
                 CUCcuucc A cu GA uccu G cTsT 
                 655 
               
               
                   
                   
               
               
                   
                 1035 
                 AGCuccuucc A cu GA uccuTsT 
                 656 
               
               
                   
                   
               
               
                   
                 1094 
                 UGG G u GG cc GGGGAGG u GG TsT 
                 657 
               
               
                   
                   
               
               
                   
                 1386 
                 ACC AGGAAGGG cuucc AG uTsT 
                 658 
               
               
                   
                   
               
               
                   
                  759 
                 HB ucAAAGuccGAGAGucAGGTT B 
                 659 
               
               
                   
                   
               
               
                   
                  759 
                 HB uc AAAG ucc GAGAG uc AGG TT B 
                 660 
               
               
                   
                   
               
               
                   
                  759 
                 B ucAAAGuccGAGAGucAGGTT B 
                 661 
               
               
                   
                   
               
               
                   
                  759 
                 B uc AAAG ucc GAGAG uc AGG TT B 
                 662 
               
               
                   
                   
               
               
                   
                  759 
                 ccu GA cucuc GGA cuuu GA TsT 
                 663 
               
               
                   
                   
               
               
                   
                  759 
                 ccuGAcucucGGAcuuuGATsT 
                 664 
               
               
                   
                   
               
               
                   
                  283 
                 B cuuccuAAGAAcAAAAAccTT B 
                 665 
               
               
                   
                   
               
               
                   
                  461 
                 B GGGAGAuGGuGuGGGAGcATT B 
                 666 
               
               
                   
                   
               
               
                   
                  463 
                 B GAGAuGGuGuGGGAGcAGATT B 
                 667 
               
               
                   
                   
               
               
                   
                  464 
                 B AGAuGGuGuGGGAGcAGAATT B 
                 668 
               
               
                   
                   
               
               
                   
                  703 
                 B AccAcAuGGccuGAcuuuGTT B 
                 669 
               
               
                   
                   
               
               
                   
                  704 
                 B ccAcAuGGccuGAcuuuGGTT B 
                 670 
               
               
                   
                   
               
               
                   
                  705 
                 B cAcAuGGccuGAcuuuGGATT B 
                 671 
               
               
                   
                   
               
               
                   
                  706 
                 B AcAuGGccuGAcuuuGGAGTT B 
                 672 
               
               
                   
                   
               
               
                   
                  707 
                 B cAuGGccuGAcuuuGGAGuTT B 
                 673 
               
               
                   
                   
               
               
                   
                  708 
                 B AuGGccuGAcuuuGGAGucTT B 
                 674 
               
               
                   
                   
               
               
                   
                  709 
                 B uGGccuGAcuuuGGAGuccTT B 
                 675 
               
               
                   
                   
               
               
                   
                  757 
                 B cuuuucAAAGuccGAGAGuTT B 
                 676 
               
               
                   
                   
               
               
                   
                  758 
                 B uuuucAAAGuccGAGAGucTT B 
                 677 
               
               
                   
                   
               
               
                   
                  759 
                 B uuucAAAGuccGAGAGucATT B 
                 678 
               
               
                   
                   
               
               
                   
                  760 
                 B uucAAAGuccGAGAGucAGTT B 
                 679 
               
               
                   
                   
               
               
                   
                  875 
                 B uGAuGGAcAAGAGGAAAGATT B 
                 680 
               
               
                   
                   
               
               
                   
                  974 
                 B uGcGcuucuccuAccuGGcTT B 
                 681 
               
               
                   
                   
               
               
                   
                  976 
                 B cGcuucuccuAccuGGcuGTT B 
                 682 
               
               
                   
                   
               
               
                   
                  977 
                 B GcuucuccuAccuGGcuGuTT B 
                 683 
               
               
                   
                   
               
               
                   
                 1033 
                 B GuGcAGGAucAGuGGAAGGTT B 
                 684 
               
               
                   
                   
               
               
                   
                 1034 
                 B uGcAGGAucAGuGGAAGGATT B 
                 685 
               
               
                   
                   
               
               
                   
                 1036 
                 B cAGGAucAGuGGAAGGAGcTT B 
                 686 
               
               
                   
                   
               
               
                   
                 1389 
                 B cuGGAAGcccuuccuGGucTT B 
                 687 
               
               
                   
                   
               
               
                   
                 1390 
                 B uGGAAGcccuuccuGGucATT B 
                 688 
               
               
                   
                   
               
               
                   
                 1391 
                 B GGAAGcccuuccuGGucAATT B 
                 689 
               
               
                   
                   
               
               
                   
                  283 
                 B cuuccu AAGAAcAAAAA ccTT B 
                 690 
               
               
                   
                   
               
               
                   
                  461 
                 B  GGGAGA u GG u G u GGGAG c A TT B 
                 691 
               
               
                   
                   
               
               
                   
                  463 
                 B  GAGA u GG u G u GGGAG c AGA TT B 
                 692 
               
               
                   
                   
               
               
                   
                  464 
                 B  AGA u GG u G u GGGAG c AGAA TT B 
                 693 
               
               
                   
                   
               
               
                   
                  703 
                 B  A cc A c A u GG ccu GA cuuu G TT B 
                 694 
               
               
                   
                   
               
               
                   
                  704 
                 B cc A c A u GG ccu GA cuuu GG TT B 
                 695 
               
               
                   
                   
               
               
                   
                  705 
                 B c A c A u GG ccu GA cuuu GGA TT B 
                 696 
               
               
                   
                   
               
               
                   
                  706 
                 B  A c A u GG ccu GA cuuu GGAG TT B 
                 697 
               
               
                   
                   
               
               
                   
                  707 
                 B c A u GG ccu GA cuuu GGAG uTT B 
                 698 
               
               
                   
                   
               
               
                   
                  708 
                 B  A u GG ccu GA cuuu GGAG ucTT B 
                 699 
               
               
                   
                   
               
               
                   
                  709 
                 B u GG ccu GA cuuu GGAG uccTT B 
                 700 
               
               
                   
                   
               
               
                   
                  757 
                 B cuuuuc AAAG ucc GAGAG uTT B 
                 701 
               
               
                   
                   
               
               
                   
                  758 
                 B uuuuc AAAG ucc GAGAG ucTT B 
                 702 
               
               
                   
                   
               
               
                   
                  759 
                 B uuuc AAAG ucc GAGAG uc A TT B 
                 703 
               
               
                   
                   
               
               
                   
                  760 
                 B uuc AAAG ucc GAGAG uc AG TT B 
                 704 
               
               
                   
                   
               
               
                   
                  875 
                 B u GA u GGA c AAGAGGAAAGA TT B 
                 705 
               
               
                   
                   
               
               
                   
                  974 
                 B u G c G cuucuccu A ccu GG cTT B 
                 706 
               
               
                   
                   
               
               
                   
                  976 
                 B c G cuucuccu A ccu GG cu G TT B 
                 707 
               
               
                   
                   
               
               
                   
                  977 
                 B  G cuucuccu A ccu GG cu G uTT B 
                 708 
               
               
                   
                   
               
               
                   
                 1033 
                 B  G u G c AGGA uc AG u GGAAGG TT B 
                 709 
               
               
                   
                   
               
               
                   
                 1034 
                 B u G c AGGA uc AG u GGAAGGA TT B 
                 710 
               
               
                   
                   
               
               
                   
                 1036 
                 B c AGGA uc AG u GGAAGGAG cTT B 
                 711 
               
               
                   
                   
               
               
                   
                 1389 
                 B cu GGAAG cccuuccu GG ucTT B 
                 712 
               
               
                   
                   
               
               
                   
                 1390 
                 B u GGAAG cccuuccu GG uc A TT B 
                 713 
               
               
                   
                   
               
               
                   
                 1391 
                 B  GGAAG cccuuccu GG uc AA TT B 
                 714 
               
               
                   
                   
               
               
                   
                  283 
                 GGUuuuu G uucuu AGGAAG TsT 
                 715 
               
               
                   
                   
               
               
                   
                  461 
                 UGCuccc A c A cc A ucucccTsT 
                 716 
               
               
                   
                   
               
               
                   
                  463 
                 UCUGcuccc A c A cc A ucucTsT 
                 717 
               
               
                   
                   
               
               
                   
                  464 
                 UUCu G cuccc A c A cc A ucuTsT 
                 718 
               
               
                   
                   
               
               
                   
                  703 
                 CAA AG uc AGG cc A u G u GG uTsT 
                 719 
               
               
                   
                   
               
               
                   
                  704 
                 CCA AAG uc AGG cc A u G u GG TsT 
                 720 
               
               
                   
                   
               
               
                   
                  705 
                 UCC AAAG uc AGG cc A u G u G TsT 
                 721 
               
               
                   
                   
               
               
                   
                  706 
                 CUCc AAAG uc AGG cc A u G uTsT 
                 722 
               
               
                   
                   
               
               
                   
                  707 
                 ACUcc AAAG uc AGG cc A u G TsT 
                 723 
               
               
                   
                   
               
               
                   
                  708 
                 GACucc AAAG uc AGG cc A uTsT 
                 724 
               
               
                   
                   
               
               
                   
                  709 
                 GGAcucc AAAG uc AGG cc A TsT 
                 725 
               
               
                   
                   
               
               
                   
                  757 
                 ACUcuc GGA cuuu GAAAAG TsT 
                 726 
               
               
                   
                   
               
               
                   
                  758 
                 GACucuc GGA cuuu GAAAA TsT 
                 727 
               
               
                   
                   
               
               
                   
                  759 
                 UGAcucuc GGA cuuu GAAA TsT 
                 728 
               
               
                   
                   
               
               
                   
                  760 
                 CUG A cucuc GGA cuuu GAA TsT 
                 729 
               
               
                   
                   
               
               
                   
                  875 
                 UCUuuccucuu G ucc A uc A TsT 
                 730 
               
               
                   
                   
               
               
                   
                  974 
                 GCC AGG u AGGAGAAG c G c A TsT 
                 731 
               
               
                   
                   
               
               
                   
                  976 
                 CAGcc AGG u AGGAGAAG c G TsT 
                 732 
               
               
                   
                   
               
               
                   
                  977 
                 ACA G cc AGG u AGGAGAAG cTsT 
                 733 
               
               
                   
                   
               
               
                   
                 1033 
                 CCUucc A cu GA uccu G c A cTsT 
                 734 
               
               
                   
                   
               
               
                   
                 1034 
                 UCCuucc A cu GA uccu G c A TsT 
                 735 
               
               
                   
                   
               
               
                   
                 1036 
                 GCUccuucc A cu GA uccu G TsT 
                 736 
               
               
                   
                   
               
               
                   
                 1389 
                 GACc AGGAAGGG cuucc AG TsT 
                 737 
               
               
                   
                   
               
               
                   
                 1390 
                 UGAcc AGGAAGGG cuucc A TsT 
                 738 
               
               
                   
                   
               
               
                   
                 1391 
                 UUG A cc AGGAAGGG cuuccTsT 
                 739 
               
               
                   
                   
               
               
                   
                 Uppercase = ribonucleotide 
               
               
                   
                 u, c = 2′-deoxy-2′-fluoro U, C 
               
               
                   
                 T = thymidine 
               
               
                   
                 B = inverted deoxy abasic 
               
               
                   
                 s = phosphorothioate linkage 
               
               
                   
                   A  = deoxy Adenosine 
               
               
                   
                   G  = deoxy Guanosine 
               
               
                   
                   G  = 2′-O-methyl Guanosine 
               
               
                   
                   A  = 2′-O-methyl Adenosine 
               
               
                   
                 H = conjugated branched linker dicholesterol moiety 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE IV 
               
             
            
               
                   
               
               
                 Non-limiting examples of Stabilization Chemistries 
               
               
                 for chemically modified siNA constructs 
               
            
           
           
               
               
               
               
               
               
            
               
                 Chemistry 
                 pyrimidine 
                 Purine 
                 cap 
                 p = S 
                 Strand 
               
               
                   
               
               
                 “Stab 00” 
                 Ribo 
                 Ribo 
                 TT at 3′-ends 
                   
                 S/AS 
               
               
                 “Stab 1” 
                 Ribo 
                 Ribo 
                 — 
                 5 at 5′-end 
                 S/AS 
               
               
                   
                   
                   
                   
                 1 at 3′-end 
               
               
                 “Stab 2” 
                 Ribo 
                 Ribo 
                 — 
                 All linkages 
                 Usually AS 
               
               
                 “Stab 3” 
                 2′-fluoro 
                 Ribo 
                 — 
                 4 at 5′-end 
                 Usually S 
               
               
                   
                   
                   
                   
                 4 at 3′-end 
               
               
                 “Stab 4” 
                 2′-fluoro 
                 Ribo 
                 5′ and 3′-ends 
                 — 
                 Usually S 
               
               
                 “Stab 5” 
                 2′-fluoro 
                 Ribo 
                 — 
                 1 at 3′-end 
                 Usually AS 
               
               
                 “Stab 6” 
                 2′-O-Methyl 
                 Ribo 
                 5′ and 3′-ends 
                 — 
                 Usually S 
               
               
                 “Stab 7” 
                 2′-fluoro 
                 2′-deoxy 
                 5′ and 3′-ends 
                 — 
                 Usually S 
               
               
                 “Stab 8” 
                 2′-fluoro 
                 2′-O-Methyl 
                 — 
                 1 at 3′-end 
                 S/AS 
               
               
                 “Stab 9” 
                 Ribo 
                 Ribo 
                 5′ and 3′-ends 
                 — 
                 Usually S 
               
               
                 “Stab 10” 
                 Ribo 
                 Ribo 
                 — 
                 1 at 3′-end 
                 Usually AS 
               
               
                 “Stab 11” 
                 2′-fluoro 
                 2′-deoxy 
                 — 
                 1 at 3′-end 
                 Usually AS 
               
               
                 “Stab 12” 
                 2′-fluoro 
                 LNA 
                 5′ and 3′-ends 
                   
                 Usually S 
               
               
                 “Stab 13” 
                 2′-fluoro 
                 LNA 
                   
                 1 at 3′-end 
                 Usually AS 
               
               
                 “Stab 14” 
                 2′-fluoro 
                 2′-deoxy 
                   
                 2 at 5′-end 
                 Usually AS 
               
               
                   
                   
                   
                   
                 1 at 3′-end 
               
               
                 “Stab 15” 
                 2′-deoxy 
                 2′-deoxy 
                   
                 2 at 5′-end 
                 Usually AS 
               
               
                   
                   
                   
                   
                 1 at 3′-end 
               
               
                 “Stab 16” 
                 Ribo 
                 2′-O-Methyl 
                 5′ and 3′-ends 
                   
                 Usually S 
               
               
                 “Stab 17” 
                 2′-O-Methyl 
                 2′-O-Methyl 
                 5′ and 3′-ends 
                   
                 Usually S 
               
               
                 “Stab 18” 
                 2′-fluoro 
                 2′-O-Methyl 
                 5′ and 3′-ends 
                   
                 Usually S 
               
               
                 “Stab 19” 
                 2′-fluoro 
                 2′-O-Methyl 
                 3′-end 
                   
                 S/AS 
               
               
                 “Stab 20” 
                 2′-fluoro 
                 2′-deoxy 
                 3′-end 
                   
                 Usually AS 
               
               
                 “Stab 21” 
                 2′-fluoro 
                 Ribo 
                 3′-end 
                   
                 Usually AS 
               
               
                 “Stab 22” 
                 Ribo 
                 Ribo 
                 3′-end 
                   
                 Usually AS 
               
               
                 “Stab 23” 
                 2′-fluoro* 
                 2′-deoxy* 
                 5′ and 3′-ends 
                   
                 Usually S 
               
               
                 “Stab 24” 
                 2′-fluoro* 
                 2′-O-Methyl* 
                 — 
                 1 at 3′-end 
                 S/AS 
               
               
                 “Stab 25” 
                 2′-fluoro* 
                 2′-O-Methyl* 
                 — 
                 1 at 3′-end 
                 S/AS 
               
               
                 “Stab 26” 
                 2′-fluoro* 
                 2′-O-Methyl* 
                 — 
                   
                 S/AS 
               
               
                 “Stab 27” 
                 2′-fluoro* 
                 2′-O-Methyl* 
                 3′-end 
                   
                 S/AS 
               
               
                 “Stab 28” 
                 2′-fluoro* 
                 2′-O-Methyl* 
                 3′-end 
                   
                 S/AS 
               
               
                 “Stab 29” 
                 2′-fluoro* 
                 2′-O-Methyl* 
                   
                 1 at 3′-end 
                 S/AS 
               
               
                 “Stab 30” 
                 2′-fluoro* 
                 2′-O-Methyl* 
                   
                   
                 S/AS 
               
               
                 “Stab 31” 
                 2′-fluoro* 
                 2′-O-Methyl* 
                 3′-end 
                   
                 S/AS 
               
               
                 “Stab 32” 
                 2′-fluoro 
                 2′-O-Methyl 
                   
                   
                 S/AS 
               
               
                 “Stab 33” 
                 2′-fluoro 
                 2′-deoxy* 
                 5′ and 3′-ends 
                 — 
                 Usually S 
               
               
                 “Stab 34” 
                 2′-fluoro 
                 2′-O-Methyl* 
                 5′ and 3′-ends 
                   
                 Usually S 
               
               
                 “Stab 3F” 
                 2′-OCF3 
                 Ribo 
                 — 
                 4 at 5′-end 
                 Usually S 
               
               
                   
                   
                   
                   
                 4 at 3′-end 
               
               
                 “Stab 4F” 
                 2′-OCF3 
                 Ribo 
                 5′ and 3′-ends 
                 — 
                 Usually S 
               
               
                 “Stab 5F” 
                 2′-OCF3 
                 Ribo 
                 — 
                 1 at 3′-end 
                 Usually AS 
               
               
                 “Stab 7F” 
                 2′-OCF3 
                 2′-deoxy 
                 5′ and 3′-ends 
                 — 
                 Usually S 
               
               
                 “Stab 8F” 
                 2′-OCF3 
                 2′-O-Methyl 
                 — 
                 1 at 3′-end 
                 S/AS 
               
               
                 “Stab 11F” 
                 2′-OCF3 
                 2′-deoxy 
                 — 
                 1 at 3′-end 
                 Usually AS 
               
               
                 “Stab 12F” 
                 2′-OCF3 
                 LNA 
                 5′ and 3′-ends 
                   
                 Usually S 
               
               
                 “Stab 13F” 
                 2′-OCF3 
                 LNA 
                   
                 1 at 3′-end 
                 Usually AS 
               
               
                 “Stab 14F” 
                 2′-OCF3 
                 2′-deoxy 
                   
                 2 at 5′-end 
                 Usually AS 
               
               
                   
                   
                   
                   
                 1 at 3′-end 
               
               
                 “Stab 15F” 
                 2′-OCF3 
                 2′-deoxy 
                   
                 2 at 5′-end 
                 Usually AS 
               
               
                   
                   
                   
                   
                 1 at 3′-end 
               
               
                 “Stab 18F” 
                 2′-OCF3 
                 2′-O-Methyl 
                 5′ and 3′-ends 
                   
                 Usually S 
               
               
                 “Stab 19F” 
                 2′-OCF3 
                 2′-O-Methyl 
                 3′-end 
                   
                 S/AS 
               
               
                 “Stab 20F” 
                 2′-OCF3 
                 2′-deoxy 
                 3′-end 
                   
                 Usually AS 
               
               
                 “Stab 21F” 
                 2′-OCF3 
                 Ribo 
                 3′-end 
                   
                 Usually AS 
               
               
                 “Stab 23F” 
                 2′-OCF3* 
                 2′-deoxy* 
                 5′ and 3′-ends 
                   
                 Usually S 
               
               
                 “Stab 24F” 
                 2′-OCF3* 
                 2′-O-Methyl* 
                 — 
                 1 at 3′-end 
                 S/AS 
               
               
                 “Stab 25F” 
                 2′-OCF3* 
                 2′-O-Methyl* 
                 — 
                 1 at 3′-end 
                 S/AS 
               
               
                 “Stab 26F” 
                 2′-OCF3* 
                 2′-O-Methyl* 
                 — 
                   
                 S/AS 
               
               
                 “Stab 27F” 
                 2′-OCF3* 
                 2′-O-Methyl* 
                 3′-end 
                   
                 S/AS 
               
               
                 “Stab 28F” 
                 2′-OCF3* 
                 2′-O-Methyl* 
                 3′-end 
                   
                 S/AS 
               
               
                 “Stab 29F” 
                 2′-OCF3* 
                 2′-O-Methyl* 
                   
                 1 at 3′-end 
                 S/AS 
               
               
                 “Stab 30F” 
                 2′-OCF3* 
                 2′-O-Methyl* 
                   
                   
                 S/AS 
               
               
                 “Stab 31F” 
                 2′-OCF3* 
                 2′-O-Methyl* 
                 3′-end 
                   
                 S/AS 
               
               
                 “Stab 32F” 
                 2′-OCF3 
                 2′-O-Methyl 
                   
                   
                 S/AS 
               
               
                 “Stab 33F” 
                 2′-OCF3 
                 2′-deoxy* 
                 5′ and 3′-ends 
                 — 
                 Usually S 
               
               
                 “Stab 34F” 
                 2′-OCF3 
                 2′-O-Methyl* 
                 5′ and 3′-ends 
                   
                 Usually S 
               
               
                   
               
               
                 CAP = any terminal cap, see for example FIG. 10. 
               
               
                 All Stab 00-34 chemistries can comprise 3′-terminal thymidine (TT) residues 
               
               
                 All Stab 00-34 chemistries typically comprise about 21 nucleotides, but can vary as described herein. 
               
               
                 S = sense strand 
               
               
                 AS = antisense strand 
               
               
                 *Stab 23 has a single ribonucleotide adjacent to 3′-CAP 
               
               
                 *Stab 24 and Stab 28 have a single ribonucleotide at 5′-terminus 
               
               
                 *Stab 25, Stab 26, and Stab 27 have three ribonucleotides at 5′-terminus 
               
               
                 *Stab 29, Stab 30, Stab 31, Stab 33, and Stab 34 any purine at first three nucleotide positions from 5′-terminus are ribonucleotides 
               
               
                 p = phosphorothioate linkage 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE V 
               
               
                   
               
             
            
               
                 A. 2.5 μmol Synthesis Cycle ABI 394 Instrument 
               
            
           
           
               
               
               
               
               
               
            
               
                 Reagent 
                 Equivalents 
                 Amount 
                 Wait Time* DNA 
                 Wait Time* 2′-O-methyl 
                 Wait Time*RNA 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                 Phosphoramidites 
                 6.5 
                 163 
                 μL 
                 45 
                 sec 
                 2.5 
                 min 
                 7.5 
                 min 
               
               
                 S-Ethyl Tetrazole 
                 23.8 
                 238 
                 μL 
                 45 
                 sec 
                 2.5 
                 min 
                 7.5 
                 min 
               
               
                 Acetic Anhydride 
                 100 
                 233 
                 μL 
                 5 
                 sec 
                 5 
                 sec 
                 5 
                 sec 
               
               
                 N-Methyl 
                 186 
                 233 
                 μL 
                 5 
                 sec 
                 5 
                 sec 
                 5 
                 sec 
               
               
                 Imidazole 
               
               
                 TCA 
                 176 
                 2.3 
                 mL 
                 21 
                 sec 
                 21 
                 sec 
                 21 
                 sec 
               
               
                 Iodine 
                 11.2 
                 1.7 
                 mL 
                 45 
                 sec 
                 45 
                 sec 
                 45 
                 sec 
               
               
                 Beaucage 
                 12.9 
                 645 
                 μL 
                 100 
                 sec 
                 300 
                 sec 
                 300 
                 sec 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Acetonitrile 
                 NA 
                 6.67 
                 mL 
                 NA 
                 NA 
                 NA 
               
               
                   
               
            
           
           
               
            
               
                 B. 0.2 μmol Synthesis Cycle ABI 394 Instrument 
               
            
           
           
               
               
               
               
               
               
            
               
                 Reagent 
                 Equivalents 
                 Amount 
                 Wait Time* DNA 
                 Wait Time* 2′-O-methyl 
                 Wait Time*RNA 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                 Phosphoramidites 
                 15 
                 31 
                 μL 
                 45 
                 sec 
                 233 
                 sec 
                 465 
                 sec 
               
               
                 S-Ethyl Tetrazole 
                 38.7 
                 31 
                 μL 
                 45 
                 sec 
                 233 
                 min 
                 465 
                 sec 
               
               
                 Acetic Anhydride 
                 655 
                 124 
                 μL 
                 5 
                 sec 
                 5 
                 sec 
                 5 
                 sec 
               
               
                 N-Methyl 
                 1245 
                 124 
                 μL 
                 5 
                 sec 
                 5 
                 sec 
                 5 
                 sec 
               
               
                 Imidazole 
               
               
                 TCA 
                 700 
                 732 
                 μL 
                 10 
                 sec 
                 10 
                 sec 
                 10 
                 sec 
               
               
                 Iodine 
                 20.6 
                 244 
                 μL 
                 15 
                 sec 
                 15 
                 sec 
                 15 
                 sec 
               
               
                 Beaucage 
                 7.7 
                 232 
                 μL 
                 100 
                 sec 
                 300 
                 sec 
                 300 
                 sec 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Acetonitrile 
                 NA 
                 2.64 
                 mL 
                 NA 
                 NA 
                 NA 
               
               
                   
               
            
           
           
               
            
               
                 C. 0.2 μmol Synthesis Cycle 96 well Instrument 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                 Equivalents: DNA/ 
                 Amount: DNA/2′-O- 
                   
                 Wait Time* 2′-O- 
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 Reagent 
                 2′-O-methyl/Ribo 
                 methyl/Ribo 
                 Wait Time* DNA 
                 methyl 
                 Wait Time* Ribo 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                 Phosphoramidites 
                 22/33/66 
                 40/60/120 
                 μL 
                 60 
                 sec 
                 180 
                 sec 
                 360 
                 sec 
               
               
                 S-Ethyl Tetrazole 
                  70/105/210 
                 40/60/120 
                 μL 
                 60 
                 sec 
                 180 
                 min 
                 360 
                 sec 
               
               
                 Acetic Anhydride 
                 265/265/265 
                 50/50/50 
                 μL 
                 10 
                 sec 
                 10 
                 sec 
                 10 
                 sec 
               
               
                 N-Methyl 
                 502/502/502 
                 50/50/50 
                 μL 
                 10 
                 sec 
                 10 
                 sec 
                 10 
                 sec 
               
               
                 Imidazole 
               
               
                 TCA 
                 238/475/475 
                 250/500/500 
                 μL 
                 15 
                 sec 
                 15 
                 sec 
                 15 
                 sec 
               
               
                 Iodine 
                 6.8/6.8/6.8 
                 80/80/80 
                 μL 
                 30 
                 sec 
                 30 
                 sec 
                 30 
                 sec 
               
               
                 Beaucage 
                 34/51/51 
                 80/120/120 
                   
                 100 
                 sec 
                 200 
                 sec 
                 200 
                 sec 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Acetonitrile 
                 NA 
                 1150/1150/1150 
                 μL 
                 NA 
                 NA 
                 NA 
               
               
                   
               
               
                 Wait time does not include contact time during delivery. 
               
               
                 Tandem synthesis utilizes double coupling of linker molecule