RNAi induced huntingtin gene suppression
The present invention provides for a double stranded RNA comprising a first RNA sequence and a second RNA sequence wherein the first and second RNA sequence are substantially complementary, wherein the first RNA sequence has a sequence length of at least 19 nucleotides and is substantially complementary to SEQ ID NO. 1. Said double stranded RNA is for use in inducing RNAi against Huntingtin exon 1 sequences. The double stranded RNA of to the invention was capable of reducing neuronal cell death and huntingtin aggregates in an animal model.
1. A method of reducing expression of an RNA transcript comprising SEQ ID NO:1 in a neuron, method comprising introducing into the neuron a double stranded RNA comprising a first RNA sequence and a second RNA sequence wherein the first and second RNA sequence are substantially complementary, wherein the first RNA sequence comprises SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7.
2. The method of claim 1 , wherein the double stranded RNA is comprised in a pre-miRNA scaffold, a pri-miRNA scaffold, a shRNA, or an siRNA.
3. The method of claim 1 , wherein the first RNA sequence comprises 19-27 nucleotides.
4. The method of claim 1 , wherein the double stranded RNA is encoded by an expression cassette.
5. The method of claim 4 , wherein the expression cassette comprises a neuron-specific promoter.
6. The method of claim 1 , wherein the first strand and second strand are respectively selected from the group consisting of SEQ ID NOs: 3 and 8; SEQ ID NOs: 4 and 9; SEQ ID NOs: 5 and 10; SEQ ID NOs: 5 and 13; SEQ ID NOs: 5 and 14; SEQ ID NOs: 6 and 11; and SEQ ID NOs: and 7 and 12.
7. The method of claim 1 , wherein the double stranded RNA is comprised in a pre-miRNA scaffold derived from miR-451a or miR-155.
8. The method of claim 1 , wherein the double stranded RNA is introduced into the neuron through transduction by a viral vector.
9. The method of claim 8 , wherein the viral vector is an adeno associated viral (AAV) vector.
10. The method of claim 9 , wherein the AAV vector is a serotype 5 vector.
11. A method of treating a subject with Huntington's disease or a subject that is genetically predisposed for Huntington's disease who does not yet show symptoms of Huntington's disease, comprising administering to a subject with Huntington's disease an effective amount of a viral vector encoding a double stranded RNA comprising a first RNA sequence and a second RNA sequence wherein the first and second RNA sequence are substantially complementary, wherein the first RNA sequence comprises SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or SEQ ID NO:7.
12. The method of claim 11 , wherein the viral vector is an adeno associated viral (AAV) vector.
13. The method of claim 11 , wherein the AAV vector is a serotype 5 vector.
14. The method of claim 11 , wherein the double stranded RNA is comprised in a pre-miRNA scaffold, a pri-miRNA scaffold, a shRNA, or an siRNA.
15. The method of claim 11 , wherein the viral vector is administered directly into the central nervous system (CNS).
16. The method of claim 15 , wherein administration in the CNS comprises intrathecal infusion or injection into at least one of the striatum and thalamus.
17. The method of claim 11 , wherein the double stranded RNA is encoded by an expression cassette in the viral vector.
18. The method of claim 17 , wherein the expression cassette comprises a neuron-specific promoter.