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 non-human animal comprising a polynucleotide encoding a fusion protein comprising a functional domain and an engineered zinc finger polypeptide, the engineered zinc finger polypeptide comprising at least two zinc finger DNA binding domains, wherein at least one zinc finger DNA binding domain comprises a non-naturally occurring recognition helix that has been engineered to bind to a target nucleotide sequence, wherein the engineered zinc finger polypeptide binds to the target nucleotide sequence in a cell in the animal and modulates the expression of a target gene, wherein the target gene comprises an EPO gene or a receptor gene. 
     
     
         2 . The non-human animal of  claim 1 , wherein the polynucleotide is stably integrated into the genome of the cell. 
     
     
         3 . The non-human animal according to  claim 1  in which the target gene is an endogenous gene. 
     
     
         4 . The non-human animal according to  claim 1 , in which the target gene comprises a heterologous gene which is introduced into the cell or an ancestor of that cell. 
     
     
         5 . The non-human animal according to  claim 1 , in which the zinc finger polypeptide binds to a promoter or other control sequence of the target gene to modulate its expression. 
     
     
         6 . The non-human animal according to  claim 1 , wherein the zinc finger polypeptide is operably linked to an inducible promoter. 
     
     
         7 . The non-human animal according to  claim 1 , wherein the functional domain comprises a transcriptional activation domain. 
     
     
         8 . The non-human animal according to  claim 1 , wherein the functional domain comprises a transcriptional repression domain. 
     
     
         9 . The non-human animal according to  claim 1 , wherein the functional domain comprises an endonuclease. 
     
     
         10 . A method of determining the function of a gene, the method comprising the steps of
 (a) providing an animal according to  claim 1 ; and   (b) observing a phenotype of the animal as compared to an animal not comprising the transgene, thereby determining the function of the target gene.   
     
     
         11 . A method of identifying a molecule which modulates the interaction between a zinc finger polypeptide and a target nucleic acid sequence, the method comprising the steps of
 (a) providing an animal according to  claim 1 ;   (b) exposing the animal to a candidate molecule; and   (c) detecting binding or modulation of binding between the zinc finger polypeptide and the target nucleic acid sequence.