Claims
- 1. A method for modulating expression of an endogenous gene in a cell, wherein the method comprises:
a) administering to the cell a nucleic acid molecule comprising a polynucleotide sequence encoding an engineered zinc finger protein, wherein the engineered zinc finger protein comprises a plurality of zinc fingers and further wherein the amino acid sequence of at least one of the zinc fingers is obtained from the information in a database of existing zinc finger proteins and their respective binding affinities; and b) maintaining the cell under conditions in which the engineered zinc finger protein is expressed and binds to a target site in the gene.
- 2. The method of claim 1, wherein the endogenous gene is the VEGF gene.
- 3. The method of claim 1, wherein the endogenous gene is a microbial gene.
- 4. The method of claim 1,wherein the endogenous gene is a viral gene.
- 5. The method of claim 1, wherein the cell is selected from the group consisting of animal cells, plant cells, bacterial cells, protozoal cells and fungal cells.
- 6. The method of claim 5, wherein the cell is a mammalian cell.
- 7. The method of claim 6, wherein the cell is a human cell.
- 8. The method of claim 1, wherein the zinc finger protein further comprises a regulatory domain.
- 9. The method of claim 8, wherein the regulatory domain is an activation domain.
- 10. The method of claim 9, wherein the regulatory domain is selected from the group consisting of VP16, VP64 and the p65 subunit of nuclear factor kappa B.
- 11. The method of claim 8, wherein the regulatory domain is a repression domain.
- 12. A method for modulating expression of an endogenous gene in a cell, wherein the method comprises:
a) administering to the cell a nucleic acid molecule comprising a polynucleotide sequence encoding an engineered zinc finger protein, wherein the engineered zinc finger protein comprises a plurality of zinc fingers and further wherein each zinc finger is obtained from a preexisting zinc finger protein; and b) maintaining the cell under conditions in which the engineered zinc finger protein is expressed and binds to a target site in the gene.
- 13. The method of claim 12, wherein the endogenous gene is the VEGF gene.
- 14. The method of claim 12, wherein the endogenous gene is a microbial gene.
- 15. The method of claim 12, wherein the endogenous gene is a viral gene.
- 16. The method of claim 12, wherein the cell is selected from the group consisting of animal cells, plant cells, bacterial cells, protozoal cells and fungal cells.
- 17. The method of claim 16, wherein the cell is a mammalian cell.
- 18. The method of claim 17, wherein the cell is a human cell.
- 19. The method of claim 12, wherein the zinc finger protein further comprises a regulatory domain.
- 20. The method of claim 19, wherein the regulatory domain is an activation domain.
- 21. The method of claim 20, wherein the regulatory domain is selected from the group consisting of VP16, VP64 and the p65 subunit of nuclear factor kappa B.
- 22. The method of claim 19, wherein the regulatory domain is a repression domain.
- 23. A method for modulating expression of an endogenous gene in a cell, wherein the method comprises:
a) administering to the cell a nucleic acid molecule comprising a polynucleotide sequence encoding an engineered zinc finger protein, wherein the zinc finger protein comprises a plurality of zinc fingers and further wherein the zinc finger protein is partially randomized; and b) maintaining the cell under conditions in which the engineered zinc finger protein is expressed and binds to a target site in the gene.
- 24. The method of claim 23, wherein the zinc finger protein comprises at least one finger, or part of a finger, from a known zinc finger protein randomly combined with another heterologous finger, or part of a finger, from a known zinc finger protein.
- 25. The method of claim 24, wherein the endogenous cellular gene is the VEGF gene.
- 26. The method of claim 24, wherein the endogenous gene is a microbial gene.
- 27. The method of claim 24, wherein the endogenous gene is a viral gene.
- 28. The method of claim 24, wherein the cell is selected from the group consisting of animal cells, plant cells, bacterial cells, protozoal cells and fungal cells.
- 29. The method of claim 28, wherein the cell is a mammalian cell.
- 30. The method of claim 29, wherein the cell is a human cell.
- 31. The method of claim 24, wherein the zinc finger protein further comprises a regulatory domain.
- 32. The method of claim 31, wherein the regulatory domain is an activation domain.
- 33. The method of claim 32, wherein the regulatory domain is selected from the group consisting of VP16, VP64 and the p65 subunit of NF kappa B.
- 34. The method of claim 31, wherein the regulatory domain is a repression domain.
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] This application is a continuation-in-part of copending U.S. patent application Ser. No. 09/229,007, filed Jan. 12, 1999, Ser. No. 09/229,037, filed Jan. 12, 1999, and Ser. No. 09/731,558, filed Dec. 6, 2000 (Ser. No. 09/731,558 being itself a continuation-in-part of U.S. patent application Ser. No. 09/456,100, filed Dec. 6, 1999, now abandoned). The present application claims priority under 35 U.S.C. § 120 to all of the aforementioned applications, the disclosures of which are hereby incorporated by reference in their entireties.
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH AND DEVELOPMENT
[0002] Certain portions of the present disclosure were made with government support under Grant No. 1 R43 DK52251-01, awarded by the National Institutes of Health. The government may have certain rights in this invention.
Continuation in Parts (4)
|
Number |
Date |
Country |
Parent |
09229007 |
Jan 1999 |
US |
Child |
10245415 |
Sep 2002 |
US |
Parent |
09229037 |
Jan 1999 |
US |
Child |
10245415 |
Sep 2002 |
US |
Parent |
09731558 |
Dec 2000 |
US |
Child |
10245415 |
Sep 2002 |
US |
Parent |
09456100 |
Dec 1999 |
US |
Child |
09731558 |
Dec 2000 |
US |