Rna interference based methods and compositions for inhibiting hbv gene expression

CLAIMS 1. A method for attenuating expression of target genes of hepatitis B virus (HBV) in a host cell, which method comprises providing a double stranded RNA (dsRNA) sequence in a host cell in an amount sufficient to attenuate expression of HBV target genes, said dsRNA sequence comprising a nucleotide sequence that hybridizes under stringent conditions to a nucleotide sequence of a HBV target gene.

2. The method of claim 1, wherein the dsRNA sequence is provided in the host cell by introducing a dsRNA sequence into the host cell directly.

3. The method of claim 1, wherein the dsRNA sequence is provided in the host cell by introducing a double stranded DNA (dsDNA) sequence into the host cell and the dsDNA sequence is transcribed into a dsRNA sequence in the host cell.

4. The method of claim 3, wherein the dsRNA sequence is introduced into the host cell via a plasmid comprising the dsDNA sequence.

5. The method of claim 1, wherein the host cell is an isolated cell or a cultured cell.

6. The method of claim 1, wherein the host cell is comprised in a whole mammal.

7. The method of claim 6, which is used to prevent or treat HBV infection in the mammal.

8. The method of claim 7, wherein the mammal is a non-human mammal.

9. The method of claim 7, wherein the mammal is a human.

10. The method of claim 1, wherein the HBV target gene is selected from the group consisting of pre-Surface 1, per-Surface 2 and surface genes (encoding HBsAg); pre-Core and Core genes (encoding HBeAg); Core gene (encoding HBcAg) ; P gene (encoding polymerase); and X gene.

11. The method of claim 1, wherein the nucleotide sequence of the HBV target gene is selected from the group consisting of SEQ ID NO : 5, SEQ ID NO : 6, SEQ ID NO : 7, SEQ ID NO : 8, SEQ ID NO : 9, SEQ ID NO : 10, and SEQ ID NO : 11.

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12. The method of claim 1, wherein the dsRNA sequence comprises a nucleotide sequence that hybridizes under stringent conditions to a nucleotide sequence set forth in SEQ ID NO : 5, SEQ ID NO : 6, SEQ ID NO : 7, SEQ ID NO : 8, SEQ ID NO : 9, SEQ ID NO : 10, and SEQ ID NO : 11.

13. The method of claim 1, wherein the dsRNA sequence comprises nucleotide sequences selected from the group consisting of siHBV-1, siHBV-2, siHBV-3, siHBV-4, siHBV-5, siHBV-6, and siHBV-7 set forth in Table 3.

14. The method of claim 1, wherein the dsRNA sequence does not comprise nucleotide sequences set forth in Table 4.

15. The method of claim 4, wherein the plasmid comprising the dsDNA sequence further comprises a nucleotide sequence encoding a detectable marker in the host cell.

16. The method of claim 15, wherein the detectable marker is a green fluorescent protein (GFP).

17. The method of claim 1, wherein the dsRNA sequence comprises at least about 19 to about 21 nucleotides in length of the target gene sequence or a complementary to the target gene sequence.

18. The method of claim 17, wherein the dsRNA sequence forms a hairpin structure.

19. The method of claim 1, wherein expression of the HBV target gene is attenuated by at least about 80%.

20. A pharmaceutical composition for preventing or treating HBV infection, which pharmaceutical composition comprises: a) a double stranded RNA (dsRNA) sequence in an amount sufficient to attenuate expression of HBV target genes in a host cell, said dsRNA sequence comprising a nucleotide sequence that hybridizes under stringent conditions to a nucleotide sequence of a HBV target gene; or b) a double stranded DNA (dsDNA) sequence in an amount sufficient to attenuate expression of HBV target genes, said dsDNA sequence comprising a nucleotide

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sequence that is transcribed in a host cell to become a dsRNA sequence that hybridizes under stringent conditions to a nucleotide sequence of a HBV target gene.

21. The pharmaceutical composition of claim 20, further comprising a pharmaceutically acceptable carrier or excipient.

22. A kit for preventing or treating HBV infection ; which kit comprises, a pharmaceutical composition of claim 20 and an instruction for using said pharmaceutical composition for preventing or treating HBV infection in a mammal.

23. An isolated oligonucleotide sequence, which: a) hybridizes, under high stringency, with an oligonucleotide sequence, or a complementary strand thereof, that is set forth in Tables 3 ; or b) has at least 90% identity to an oligonucleotide sequence, or a complementary strand thereof, that is set forth in Tables 3.

24. The isolated oligonucleotide sequence of claim 23, which comprises DNA, RNA, PNA or a derivative thereof.

25. The isolated oligonucleotide sequence of claim 23, which comprises a nucleotide sequence, or a complementary strand thereof, that is set forth in Tables 3.

26. An isolated double stranded oligonucleotide sequence, which comprises a pair of complementary oligonucleotide sequences of claim 23.

27. The isolated double stranded oligonucleotide sequence of claim 26, which is dsDNA or dsRNA.

28. A vector, which vector comprises an isolated oligonucleotide sequence of claim 23.

29. The vector of claim 28, which further comprises a nucleotide sequence encoding a detectable marker in the host cell.

30. The vector of claim 29, wherein the detectable marker is a green fluorescent protein (GFP).

31. The vector of claim 28, which is a plasmid.

32. A cell, which cell comprises a vector of claim 28.

33. The cell of claim 32, which is an isolated cell or a cultured cell.

34. The cell of claim 32, which is comprised in a non-human whole mammal.

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35. The cell of claim 32, which stably expresses encoded siRNA and/or other sequence (s).

36. The cell of claim 32, which is a HepG2. 2.15 cell.

37. A pharmaceutical composition for preventing or treating HBV infection, which pharmaceutical composition comprises an isolated oligonucleotide sequence of claim 23.

38. A pharmaceutical composition for preventing or treating HBV infection, which pharmaceutical composition comprises a vector of claim 28.