Chemically modified oligonucleotides for use in modulating micro rna and uses thereof

Abstract

This invention relates generally to chemically modified oligonucleotides useful for modulating expression of microRNAs and pre-microRNAs. More particularly, the invention relates to single stranded chemically modified oligonucleotides for inhibiting microRNA and pre-microRNA expression and to methods of making and using the modified oligonucleotides. Also included in the invention are compositions and methods for silencing microRNAs in the central nervous system.

What is claimed is: 
     
         1 . A method of reducing the amount of a microRNA in a cell in a subject comprising the step of administering an antagomir to the subject, wherein the oligonucletide agent is substantially single-stranded, comprises a sequence which is substantially complementary to 12 to 23 contiguous nucleotides of a target sequence, and wherein the target sequence differs by no more than 1, 2, or 3 nucleotides from a sequence selected from the group consisting of those provided in Table 1. 
     
     
         2 . The method of  claim 1 , wherein said antagomir is selected from the group consisting of those provided in Tables 2a-e and Table 4. 
     
     
         3 . An isolated oligonucleotide agent, comprising a nucleotide sequence sufficiently complementary to a microRNA target sequence of about 12 to 23 nucleotides, wherein the target sequence differs by no more than 1, 2, or 3 nucleotides from a sequence selected from the group consisting of those provided in Table 1. 
     
     
         4 . The antagomir of  claim 3 , comprising a nucleotide sequence sufficiently complementary to a target sequence of about 15 to 23 nucleotides of the microRNA nucleotide sequence, and wherein the target sequence differs by no more than 1, 2, or 3 nucleotides from a sequence selected from the group consisting of those provided in Table 1. 
     
     
         5 . The antagomir of  claim 3 , wherein the antagomir further comprises a non-nucleotide moiety. 
     
     
         6 . The antagomir of  claim 3 , wherein the antagomir is stabilized against nucleolytic degradation. 
     
     
         7 . The antagomir of  claim 3 , further comprising a phosphorothioate at the first internucleotide linkage at the 5′ end of the nucleotide sequence. 
     
     
         8 . The antagomir of  claim 3 , further comprising a phosphorothioate at the first internucleotide linkage at the 3′ end of the nucleotide sequence. 
     
     
         9 . The antagomir of  claim 3 , further comprising a phosphorothioate at the first internucleotide linkage at the 5′ end of the nucleotide sequence, and a phosphorothioate at the first internucleotide linkage at the 3′ end of the nucleotide sequence. 
     
     
         10 . The antagomir of  claim 3 , further comprising a 2′-modified nucleotide. 
     
     
         11 . The antagomir of  claim 10 , wherein the 2′-modified nucleotide comprises a modification selected from the group consisting of: 2′-deoxy, 2′-deoxy-2′-fluoro, 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), 2′-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), and 2′-O—N-methylacetamido (2′-O-NMA). 
     
     
         12 . The antagomir of  claim 10 , wherein the 2′-modified nucleotide comprises a 2′-O-methyl. 
     
     
         13 . The antagomir of  claim 3 , further comprising a cholesterol molecule attached to the 3′ end of the agent. 
     
     
         14 . The antagomir of  claim 3 , wherein the antagomir is selected from the group consisting of those provided in Table 2a-e and Table 4. 
     
     
         15 . A method of reducing the amount of miR-122 in a cell, comprising contacting the cell with an antagomir of  claim 3 , wherein the sequence selected from the group consisting of those provided in Table 1 is miR-122. 
     
     
         16 . A method of making an antagomir of  claim 3 , the method comprising the synthesis of the oligonucleotide agent, wherein the nucleotide sequence comprises at least one modification that stabilizes the antagomir against nucleolytic degradation. 
     
     
         17 . A pharmaceutical composition comprising an antagomir of  claim 3  and a pharmaceutically acceptable carrier. 
     
     
         18 . A method of inhibiting miR-122 expression in a cell, comprising contacting the cell with an effective amount of an antagomir of  claim 3 , wherein the sequence selected from the group consisting of those provided in Table 1 is miR-122. 
     
     
         19 . A method of increasing aldolase-A protein levels in a cell comprising contacting the cell with an effective amount of an antagomir of  claim 3 , wherein the sequence selected from the group consisting of those provided in Table 1 is miR-122.