Regulation of endogenous gene expression in cells using zinc finger proteins

Abstract

The present invention provides methods for modulating expression of endogenous cellular genes using engineered zinc finger proteins.

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 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.    
     
     
         2 . The method of  claim 1 , 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.  
     
     
         3 . The method of  claim 2 , wherein the endogenous cellular gene is the VEGF gene.  
     
     
         4 . The method of  claim 2 , wherein the endogenous gene is a microbial gene.  
     
     
         5 . The method of  claim 2 , wherein the endogenous gene is a viral gene.  
     
     
         6 . The method of  claim 2 , wherein the cell is selected from the group consisting of animal cells, plant cells, bacterial cells, protozoal cells and fungal cells.  
     
     
         7 . The method of  claim 6 , wherein the cell is a mammalian cell.  
     
     
         8 . The method of  claim 7 , wherein the cell is a human cell.  
     
     
         9 . The method of  claim 2 , wherein the zinc finger protein further comprises a regulatory domain.  
     
     
         10 . The method of  claim 9 , wherein the regulatory domain is an activation domain.  
     
     
         11 . The method of  claim 10 , wherein the regulatory domain is selected from the group consisting of VP 16, VP64 and the p65 subunit of NF kappa B.  
     
     
         12 . The method of  claim 9 , wherein the regulatory domain is a repression domain.