Foreign DNA surveillance protein
A method for augmenting expression of a heterologous nucleic acid in a eukaryotic cell or increasing the efficiency of gene expression using any gene expression system is carried out by decreasing expression or activity of an endogenous Interferon-induced protein-16 (IFI16).
1. A method of augmenting expression of a heterologous nucleic acid in a eukaryotic cell comprising:
(a) decreasing expression or activity of endogenous Interferon-induced protein-16 (IFI16) in the cell; and
(b) expressing a heterologous nucleic acid in the cell of step (a), wherein said heterologous nucleic acid comprises a viral vector comprising a non-viral or viral coding sequence;
wherein the steps of (a) and (b) do not occur at the same time.
2. The method of claim 1 , wherein said heterologous nucleic acid coding sequence comprises non-viral DNA.
3. The method of claim 1 , wherein said heterologous nucleic acid coding sequence encodes a mammalian gene product.
4. The method of claim 1 , wherein an IFI16 protein is encoded by a nucleic acid comprising the sequence of SEQ ID NO:5.
5. The method of claim 1 , wherein an IFI16 protein comprises the amino acid sequence of SEQ ID NO:1, 2, 3, or 4.
6. The method of claim 1 , wherein said expression is decreased by contacting said cell with an IFI16-specific siRNA.
7. The method of claim 6 , wherein said IFI16-specific siRNA is selected from the group consisting of GAUCUGUAAUUCAUAGUCA (SEQ ID NO: 6), GGACCAGCCCUAUCAAGAA (SEQ ID NO: 7), GGAGUAAGGUGUCCGAGGA (SEQ ID NO: 8), CAGCGUAACUCCUAAAAUC (SEQ ID NO: 9), and GCUGGUCCUAACCAAACGU (SEQ ID NO: 10).
8. The method of claim 1 , wherein said expression is decreased by contacting said cell with an IFI16-specific shRNA.
9. The method of claim 8 , wherein said IFI16-specific shRNA is selected from the group consisting of
#1 Sense:
(SEQ ID NO: 11)
CCACAAUCUACGAAAUUCA,
Anti-sense:
(SEQ ID NO: 12)
UGAAUUUCGUAGAUUGUGG;
#2 Sense:
(SEQ ID NO: 13)
CCAUCCAGCAGUUUCUUCA,
Anti-sense:
(SEQ ID NO: 14)
UGAAGAAACUGCUGGAUGG;
and
#3 Sense:
(SEQ ID NO: 15)
GGAAGGAGAUAAACUGAAA,
Anti-sense:
(SEQ ID NO: 16)
UUUCAGUUUAUCUCCUUCC.
10. The method of claim 1 , wherein said expression is decreased by contacting said cell with an IFI16-specific antisense nucleic acid.
11. The method of claim 1 , wherein said activity is decreased by contacting said cell with an inhibitor of IFI16 binding to DNA.
12. The method of claim 11 , wherein said inhibitor binds to a HIN domain of IFI16.
13. The method of claim 11 , wherein said inhibitor binds to residues K663, R667, K732, K734, K759, and/or R764 of IFI16 isotype A or a corresponding residue of isotype B or C.
14. The method of claim 6 , wherein said IFI16-regulating nucleic acid sequence comprises an IFI16-specific shRNA.
15. The method of claim 8 , wherein said inhibitor reduces binding of IFI16 to heterologous DNA or reduces IFI16-mediated addition of silencing chromatin to said heterologous DNA.
16. The method of claim 3 , wherein the eukaryotic cell is a mammalian cell.
17. The method of claim 16 , wherein the mammalian cell is a human cell.
18. The method of claim 16 , wherein the mammalian gene product is a different form of a protein than is present in the mammalian cell.
19. The method of claim 18 , wherein the protein that is present in the mammalian cell is functionally defective.
20. The method of claim 18 , wherein the protein that is present in the mammalian cell is non-functional.
21. The method of claim 18 , wherein the mammalian gene product is a normal, functionally active protein.
22. The method of claim 16 , wherein the mammalian gene product is from a different cell type compared to the mammalian cell.
23. The method of claim 16 , wherein the mammalian gene product is from a different species compared to the mammalian cell.
24. The method of claim 1 , wherein the mammalian gene product is naturally expressed by the eukaryotic cell.