siRNA-mediated gene silencing of synuclein
View Patent ↗The present invention is directed to small interfering RNAs that down regulate expression of a synuclein gene and methods of using the small interfering RNAs.
1. A method of reducing expression of a synuclein gene in a cell, said method comprising introducing into a cell a siRNA in an amount effective to down regulate expression of said synuclein gene,
wherein said siRNA comprises a nucleotide sequence has at least 90% complementarity to SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 10, or SEQ ID NO: 11 and has a length of 21-30 nucleotides.
2. The method of claim 1 wherein said nucleotide sequence is complementary to 15 to 21 consecutive nucleotides of SEQ ID NO: 7.
3. The method of claim 1 wherein said siRNA is introduced into said cell in a vector encoding said siRNA.
4. The method of claim 1 wherein said siRNA forms a hairpin structure comprising a loop.
5. A method of reducing expression of a synuclein gene in a cell, said method comprising: introducing a vector encoding a siRNA into a cell, the siRNA being introduced in an amount effective to down regulate expression of said synuclein gene, wherein said siRNA is encoded by a nucleotide sequence at least 90% identical to SEQ ID NO:21.
6. The method of claim 1 wherein said cell is a central nervous system cell or a peripheral nervous system cell.
7. A method of treating a neurodegenerative disease or a synucleinopathy in a subject comprising administering to said subject a therapeutically effective amount of a siRNA comprising a nucleotide sequence having at least 90% identity to SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 10, or SEQ ID NO: 11 and having a length of 21-30 nucleotides, wherein the expression of a synuclein gene is down regulated.
8. The method of claim 3 , wherein the vector is an expression vector comprising a promoter operably linked to the siRNA.
9. The method of claim 3 , wherein the vector is a viral vector.
10. The method of claim 8 , wherein the promoter is a constitutive promoter or a regulatable promoter operatively linked to the nucleotide sequence encoding the siRNA.
11. The method of claim 1 , wherein the siRNA comprises a 3′ overhang of at least one unpaired nucleotide.
12. The method of claim 1 , wherein the siRNA comprises a 5′ overhang of at least one unpaired nucleotide.
13. The method of claim 3 , wherein the vector is selected from the group consisting of adenoviral, lentiviral, poliovirus, adeno-associated viral, HSV, and murine Maloney-based viral vector.
14. The method of claim 3 , wherein the vector comprises a Pol III H-1 promoter operatively linked to said siRNA.
15. The method of claim 7 , wherein the neurodegenerative disease is Parkinson's disease.
16. The method of claim 7 , wherein said siRNA forms a hairpin structure comprising a loop.
17. The method of claim 7 , wherein an expression vector encodes the siRNA.