RNA interference mediated inhibition of NOGO and NOGO receptor gene expression using short interfering RNA

Abstract

The present invention concerns methods and reagents useful in modulating gene expression in a variety of applications, including use in therapeutic, diagnostic, target validation, and genomic discovery applications associated with Alzheimer's disease. Specifically, the invention relates to small interfering RNA (siRNA) molecules capable of mediating RNA interference (RNAi) against NOGO and NOGO receptor (NOGOr) polypeptide and polynucleotide targets.

1 - 36 . (canceled)  
     
     
         37 . A chemically modified double stranded short interfering nucleic acid (siNA) molecule that directs cleavage of a NOGO receptor (NOGOr) RNA via RNA interference (RNAi), wherein: 
 a. each strand of said siNA molecule is 18 to 27 nucleotides in length;    b. the antisense strand of said siNA molecule comprises a nucleotide sequence that is complementary to a nucleotide sequence of said NOGOr RNA; and the sense strand is complementary to the antisense strand; and    c. said siNA molecule comprises at least one chemically modified nucleotide or non-nucleotide at the 5′ end and/or 3′ end of the sense strand and the 3′ end of the antisense strand.    
     
     
         38 . The siNA molecule of  claim 37 , wherein said siNA molecule comprises no ribonucleotides.  
     
     
         39 . The siNA molecule of  claim 37 , wherein said siNA molecule comprises one or more ribonucleotides.  
     
     
         40 . The siNA molecule of  claim 37 , wherein said chemically modified nucleotide comprises a 2′-deoxy nucleotide.  
     
     
         41 . The siNA molecule of  claim 37 , wherein said chemically modified nucleotide comprises a 2′-deoxy-2′-fluoro nucleotide.  
     
     
         42 . The siNA molecule of  claim 37 , wherein said chemically modified nucleotide comprises a 2′-O-methyl nucleotide.  
     
     
         43 . The siNA molecule of  claim 37 , wherein said chemically modified nucleotide comprises a phosphorothioate internucleotide linkage.  
     
     
         44 . The siNA molecule of  claim 37 , wherein said non-nucleotide comprises an abasic moiety.  
     
     
         45 . The siNA molecule of  claim 44 , wherein said abasic moiety comprises an inverted deoxyabasic moiety.  
     
     
         46 . The siNA molecule of  claim 37 , wherein each strand of the siNA molecule comprises 19 to 23 nucleotides, and wherein each strand comprises at least 19 nucleotides that are complementary to the nucleotides of the other strand.  
     
     
         47 . The siNA molecule of  claim 37 , wherein said siNA molecule is assembled from two separate oligonucleotide fragments wherein one fragment comprises the sense region and a second fragment comprises the antisense region of said siNA molecule.  
     
     
         48 . The siNA molecule of  claim 37 , wherein said sense region is connected to the antisense region via a linker molecule.  
     
     
         49 . The siNA molecule of  claim 48 , wherein said linker molecule is a polynucleotide linker.  
     
     
         50 . The siNA molecule of  claim 48 , wherein said linker molecule is a non-nucleotide linker.  
     
     
         51 . The siNA molecule of  claim 37 , wherein pyrimidine nucleotides in the sense region are 2′-O-methyl pyrimidine nucleotides.  
     
     
         52 . The siNA molecule of  claim 37 , wherein purine nucleotides in the sense region are 2′-deoxy purine nucleotides.  
     
     
         53 . The siNA molecule of  claim 37 , wherein pyrimidine nucleotides present in the sense region are 2′-deoxy-2′-fluoro pyrimidine nucleotides.  
     
     
         54 . The siNA molecule of  claim 47 , wherein the fragment comprising said 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 comprising said sense region.  
     
     
         55 . The siNA molecule of  claim 54 , wherein said terminal cap moiety is an inverted deoxy abasic moiety.  
     
     
         56 . The siNA molecule of  claim 37 , wherein pyrimidine nucleotides of said antisense region are 2′-deoxy-2′-fluoro pyrimidine nucleotides  
     
     
         57 . The siNA molecule of  claim 37 , wherein purine nucleotides of said antisense region are 2′-O-methyl purine nucleotides.  
     
     
         58 . The siNA molecule of  claim 37 , wherein purine nucleotides present in said antisense region comprise 2′-deoxy-purine nucleotides.  
     
     
         59 . The siNA molecule of  claim 56 , wherein said antisense region comprises a phosphorothioate internucleotide linkage at the 3′ end of said antisense region.  
     
     
         60 . The siNA molecule of  claim 47 , wherein each of the two fragments of said siNA molecule comprise 21 nucleotides.  
     
     
         61 . The siNA molecule of  claim 60 , wherein 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 and 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.  
     
     
         62 . The siNA molecule of  claim 61 , wherein each of the two 3′ terminal nucleotides of each fragment of the siNA molecule are 2′-deoxy-pyrimidines.  
     
     
         63 . The siNA molecule of  claim 62 , wherein said 2′-deoxy-pyrimidine is 2′-deoxy-thymidine.  
     
     
         64 . The siNA molecule of  claim 60 , wherein all 21 nucleotides of each fragment of the siNA molecule are base-paired to the complementary nucleotides of the other fragment of the siNA molecule.  
     
     
         65 . The siNA molecule of  claim 60 , wherein 19 nucleotides of the antisense region are base-paired to the nucleotide sequence of the RNA encoded by a NOGOr gene or a portion thereof.  
     
     
         66 . The siNA molecule of  claim 60 , wherein 21 nucleotides of the antisense region are base-paired to the nucleotide sequence of the RNA encoded by a NOGOr gene or a portion thereof.  
     
     
         67 . The siNA molecule of  claim 47 , wherein the 5′-end of the fragment comprising said antisense region optionally includes a phosphate group.  
     
     
         68 . A pharmaceutical composition comprising the siNA molecule of  claim 37  in an acceptable carrier or diluent.