RNAi expression constructs with liver-specific enhancer/promoter
The present invention provides compositions and methods suitable for RNAi specifically in the liver so as to treat diseases or disorders.
1. A DNA directed RNA interference (ddRNAi) expression construct comprising:
one or more enhancer elements selected from the group consisting of ApoE enhancer elements and SynEnh enhancer elements;
one or more liver-specific promoters; and
one or more RNAi constructs that provide three or more RNAi agents;
wherein the RNAi agents target hepatitis virus genes that are desired to be repressed in a liver cell or whole liver, and wherein at least one of the three or more RNAi agents is coded by SEQ ID NO: 42.
2. The ddRNAi expression construct of claim 1 , wherein the expression construct comprises one liver-specific promoter and the one or more RNAi constructs provide three RNAi agents.
3. The ddRNAi expression construct of claim 1 , wherein each of the RNAi agents have different sequences.
4. The ddRNAi expression construct of claim 1 , wherein the RNAi agents are expressed as shRNAs.
5. The ddRNAi expression construct of claim 4 , further comprising nucleotides between a transcription initiation point of the expression construct and a first nucleotide of the one or more RNAi constructs.
6. The ddRNAi expression construct of claim 1 , wherein the RNAi agent is part of a primary microRNA structure.
7. The ddRNAi expression construct of claim 1 , further comprising a reporter gene.
8. The ddRNAi expression construct of claim 1 , further comprising a terminator.
9. The ddRNAi expression construct of claim 8 , further comprising a transcriptional pause site.
10. The ddRNAi expression construct of claim 1 , wherein the other RNAi agents are coded by one or more of SEQ ID NO: 1-56.
11. The ddRNAi expression construct of claim 1 , wherein the one or more promoters are selected from the group consisting of a hAAT promoter, an LCAT promoter, an ApoH promoter, and a transthyretin promoter.
12. A method of modifying the expression of one or more hepatitis virus genes in the liver, comprising delivering to the liver a DNA directed RNA interference (ddRNAi) expression construct comprising:
one or more enhancer elements selected from the group consisting of ApoE enhancer elements and SynEnh enhancer elements;
one or more liver-specific promoters; and
one or more RNAi constructs that provide three or more RNAi agents,
wherein at least one of the three or more RNAi agents is coded by SEQ ID NO: 42.
13. The ddRNAi expression construct of claim 12 , wherein the RNAi agents are expressed as shRNAs.
14. The ddRNAi expression construct of claim 13 , further comprising nucleotides between a transcription initiation point of the expression construct and a first nucleotide of the one or more RNAi constructs.
15. The ddRNAi expression construct of claim 12 , wherein the other RNAi agents are coded by one or more of SEQ ID NO: 1-56.
16. The ddRNAi expression construct of claim 12 , wherein the one or more promoters are selected from the group consisting of a hAAT promoter, an LCAT promoter, an ApoH promoter, and a transthyretin promoter.