Therapeutic compounds

Abstract

The present invention is directed to RNA interference (RNAi) molecules targeted against a Huntington's disease nucleic acid sequence, and methods of using these RNAi molecules to treat Huntington's disease.

1 - 72 . (canceled) 
     
     
         73 . A nucleic acid encoding an artificial primary miRNA transcript (pri-miRNA) comprising, in order of position, a 5′-flanking region, wherein the 5′-flanking region comprises a 5′-bulge sequence positioned upstream from a 5′-joining sequence; a non-guide region, wherein the 5′-joining sequence is contiguously linked to the non-guide region; a loop region; a guide region; and a 3′-flanking region, wherein the guide region comprises a sequence at least 80% identical to cgaccaugcgagccagca (miHDS.1 guide. SEQ ID NO:7) and the non-guide region is at least 80% complementary to the guide region. 
     
     
         74 . The nucleic acid of  claim 73 , wherein the guide region consists of 18-30 nucleotides. 
     
     
         75 . The nucleic acid of  claim 73 , wherein the 5′ joining sequence consists of 5-8 nucleotides. 
     
     
         76 . The nucleic acid of  claim 73 , wherein the 5′-bulge sequence consists of 1-10 nucleotides. 
     
     
         77 . The nucleic acid of  claim 73 , wherein the 5′-flanking region further comprises a 5′-spacer sequence positioned upstream from the 5′-bulge sequence. 
     
     
         78 . The nucleic acid of  claim 77 , wherein the 5′-spacer sequence consists of 10-12 nucleotides. 
     
     
         79 . The nucleic acid of  claim 77 , further comprising a 5′-upstream sequence positioned upstream from the 5′-spacer sequence. 
     
     
         80 . The nucleic acid of  claim 79 , wherein the 5′-upstream sequence consists of 30-2000 nucleotides. 
     
     
         81 . The nucleic acid of  claim 73 , wherein the 3′-flanking region comprises a 3′-joining sequence contiguously linked to the guide region. 
     
     
         82 . The nucleic acid of  claim 81 , wherein the 3′-joining sequence consists of 5-8 nucleotides. 
     
     
         83 . The nucleic acid of  claim 81 , wherein the 3′-joining sequence is at least about 85% complementary to the 5′-joining sequence. 
     
     
         84 . The nucleic acid of  claim 81 , further comprising a 3′-bulge sequence positioned downstream from the 3′-joining sequence. 
     
     
         85 . The nucleic acid of  claim 84 , wherein the 3′-bulge sequence consists of 1-10 nucleotides. 
     
     
         86 . The nucleic acid of  claim 84 , further comprising a 3′-spacer sequence positioned downstream from the 3′-bulge sequence. 
     
     
         87 . The nucleic acid of  claim 86 , wherein the 3′-spacer sequence consists of 10-12 nucleotides. 
     
     
         88 . The nucleic acid of  claim 86 , further comprising a 3′-downstream sequence positioned downstream from the 3′-spacer sequence. 
     
     
         89 . The nucleic acid of  claim 88 , wherein the 3′-downstream sequence is about 30-2000 nucleotides in length. 
     
     
         90 . The nucleic acid of  claim 73 , wherein the loop region is from 15-25 nucleotides in length. 
     
     
         91 . An expression cassette comprising a promoter contiguously linked to the nucleic acid of  claim 73 . 
     
     
         92 . A vector comprising the expression cassette of  claim 91 . 
     
     
         93 . The vector of  claim 90 , wherein the vector is an adeno-associated virus (AAV) vector. 
     
     
         94 . The vector of  claim 93 , wherein the AAV is AAV1, AAV2, AAV4, AAV5, or AAV2/1. 
     
     
         95 . An isolated microRNA molecule comprising the nucleic acid of  claim 73 . 
     
     
         96 . A method of inducing RNA interference comprising administering to a subject an effective amount of the nucleic acid of  claim 73 . 
     
     
         97 . A method of treating a subject with Huntington's Disease, comprising administering to the subject the nucleic acid of  claim 73  so as to treat the Huntington's Disease.