Rna interference suppression of neurodegenerative diseases and methods of use thereof

Abstract

The present invention is directed to small interfering RNA molecules (siRNA) targeted against nucleic acid sequence that encodes huntingtin or ataxin-1, and methods of using these siRNA molecules.

1 . An isolated RNA duplex comprising a first strand of RNA and a second strand of RNA, wherein the first strand comprises at least 15 contiguous nucleotides encoded by shSCA1.F10 (SEQ ID NO:102), shSCA1.F11 (SEQ ID NO:103), SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:63, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, SEQ ID NO:72, SEQ ID NO:74, SEQ ID NO:76, SEQ ID NO:78, SEQ ID NO:80, SEQ ID NO:82, SEQ ID NO:84, SEQ ID NO:86, or SEQ ID NO:88, SEQ ID NO:90, SEQ ID NO:96 through SEQ ID NO:101 or SEQ ID NO:106 through SEQ ID NO:142, and wherein the second strand is complementary to at least 12 contiguous nucleotides of the first strand. 
     
     
         2 . The RNA duplex of  claim 1 , wherein the duplex is between 15 and 30 base pairs in length. 
     
     
         3 . The RNA duplex of  claim 1 , wherein the duplex is between 19 and 25 base pairs in length. 
     
     
         4 . The RNA duplex of  claim 1 , wherein the first and/or second strand further comprises an overhang region. 
     
     
         5 . The RNA duplex of  claim 1 , wherein the first and/or second strand further comprises a 3′ overhang region, a 5′ overhang region, or both 3′ and 5′ overhang regions. 
     
     
         6 . The RNA duplex of  claim 4 , wherein the overhang region is from 1 to 10 nucleotides in length. 
     
     
         7 . The RNA duplex of  claim 1 , wherein the first strand and the second strand are operably linked by means of an RNA loop strand to form a hairpin structure comprising a duplex structure and a loop structure. 
     
     
         8 . The RNA duplex of  claim 7 , wherein the loop structure contains from 4 to 10 nucleotides. 
     
     
         9 . The RNA duplex of  claim 7 , wherein the loop structure contains 4, 5 or 6 nucleotides. 
     
     
         10 . The RNA duplex of  claim 7 , wherein the loop structure corresponds to SEQ ID NO:61 or SEQ ID NO:64. 
     
     
         11 . An expression cassette comprising a nucleic acid encoding at least one strand of the RNA duplex of  claim 1 . 
     
     
         12 . The expression cassette of  claim 11 , further comprising a promoter. 
     
     
         13 . The expression cassette of  claim 12 , wherein the promoter is a regulatable promoter. 
     
     
         14 . The expression cassette of  claim 12 , wherein the promoter is a constitutive promoter. 
     
     
         15 . The expression cassette of  claim 12 , wherein the promoter is a CMV, RSV, pol II or pol III promoter. 
     
     
         16 . The expression cassette of  claim 11 , wherein the expression cassette further comprises a polyadenylation signal. 
     
     
         17 . The expression cassette of  claim 16 , wherein the polyadenylation signal is a synthetic minimal polyadenylation signal. 
     
     
         18 . The expression cassette of  claim 11 , further comprising a marker gene. 
     
     
         19 . A vector comprising the expression cassette of  claim 11 . 
     
     
         20 - 25 . (canceled) 
     
     
         26 . A method of suppressing the accumulation of huntingtin or ataxin-1 in a cell comprising introducing a ribonucleic acid (RNA) of  claim 1  into the cell in an amount sufficient to suppress accumulation of huntingtin or ataxin-1 in the cell. 
     
     
         27 - 61 . (canceled)