RNA interference mediated inactivation of hepatitis B virus in a subject
Embodiments of the present invention provide methods for targeted inactivation of viral genomes. In one embodiment, zinc-finger proteins in which DNA binding sites are altered such that they recognize and bind different, desired DNA sequences contained in hepatitis B virus (HBV) and that include nuclease domains are used for inactivation. Other embodiments for targeted inactivation of viral genomes use small nucleic acid molecules, such as short micro-RNA molecules or short hairpin RNA molecules capable of mediating RNA interference (RNAi) against the hepatitis B virus.
1. A method for inhibiting expression of Hepatitis B virus (HBV) in a subject comprising:
delivering to the subject a precursor RNA interference trigger molecule that directs cleavage of a HBV RNA via RNA interference (RNAi) wherein:
a fully processed RNAi trigger molecule is about 19 to 29 nucleotides in length;
one strand of the fully processed RNAi trigger molecule comprises a nucleotide sequence having sufficient complementarity to the HBV RNA for the RNAi trigger molecule to direct cleavage of the HBV RNA via RNA interference;
wherein the fully processed RNAi trigger molecule is selected from SEQ ID NO: 9 through SEQ ID NO: 23; and
inhibiting expression or levels of HBV mRNA by cleavage of HBV RNA molecules in the subject therefore inhibiting HBV infection in the subject.
2. The method of claim 1 wherein the fully processed RNAi trigger molecules are incorporated into a micro RNA (miRNA).
3. The method of claim 1 wherein the fully processed RNAi trigger molecules are incorporated into short hairpin RNA (shRNA).
4. The method of claim 1 , wherein the RNAi triggers target approximately 3.5, 2.1, 2, 4, or 0.7 kilo base pair RNAs of the HBV, or any subset thereof.
5. The method of claim 1 , wherein the inhibition of HBV occurs in a mouse.
6. The method of claim 1 , wherein the inhibition of the virus HBV occurs in a human.
7. The method of claim 1 , wherein the fully processed RNAi trigger molecule is selected from one or more of SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO: 16.