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 recombinant zinc finger proteins.

What is claimed is: 
     
         1 . A method of modulating expression of an endogenous cellular gene in a cell, the method comprising the steps of:
 administering to the cell a nucleic acid molecule comprising a polynucleotide sequence which encodes a fusion molecule, the fusion molecule comprising an engineered DNA-binding domain and a regulatory domain, wherein the nucleic acid molecule expresses the fusion protein in the cell, thereby modulating expression of the endogenous cellular gene.   
     
     
         2 . The method of  claim 1 , wherein the fusion protein comprises at least two regulatory domains. 
     
     
         3 . The method of  claim 1 , wherein the cell is selected from the group consisting of animal cell, a plant cell, a bacterial cell, a protozoal cell, or a fungal cell. 
     
     
         4 . The method of  claim 3 , wherein the cell is a mammalian cell. 
     
     
         5 . The method of  claim 3 , wherein the cell is a human cell. 
     
     
         6 . The method of  claim 1 , wherein the regulatory domain is selected from the group consisting of a transcriptional repressor, a transcriptional activator, an endonuclease, a methyl transferase, and a histone deacetylase. 
     
     
         7 . The method of  claim 1 , wherein the step of administering the nucleic acid molecule to the cell comprises administering the nucleic acid molecule in a lipid:nucleic acid complex or as naked nucleic acid. 
     
     
         8 . The method of  claim 1 , wherein the nucleic acid molecule is an expression vector comprising the fusion protein-encoding nucleic acid operably linked to a promoter. 
     
     
         9 . The method of  claim 8 , wherein the expression vector is a viral expression vector. 
     
     
         10 . The method of  claim 8 , wherein the expression vector is a retroviral expression vector, an adenoviral expression vector, or an AAV expression vector. 
     
     
         11 . The method of  claim 8 , wherein the promoter is an inducible promoter. 
     
     
         12 . The method of  claim 1 , wherein the target site is upstream of a transcription initiation site of the endogenous cellular gene. 
     
     
         13 . The method of  claim 1 , wherein the target site is adjacent to a transcription initiation site of the endogenous cellular gene. 
     
     
         14 . The method of  claim 1 , wherein the target site is adjacent to an RNA polymerase pause site, wherein the RNA polymerase pause site is downstream of a transcription initiation site of the endogenous cellular gene.