Methods and compositions for gene inactivation

Abstract

Disclosed herein are methods and compositions for inactivating CCR-5 genes, using zinc finger nucleases (ZFNs) comprising a zinc finger protein and a cleavage domain or cleavage half-domain. Polynucleotides encoding ZFNs, vectors comprising polynucleotides encoding ZFNs, such as adenovirus (Ad) vectors, and cells comprising polynucleotides encoding ZFNs and/or cells comprising ZFNs are also provided.

1 . A method for inactivating an HIV receptor gene in a subject, the method comprising:
 introducing, into a cell of the subject, a homing endonuclease that binds to a target site in an HIV receptor gene and cleaves the HIV receptor gene.   
     
     
         2 . The method of  claim 1 , wherein the homing endonuclease is engineered to bind to the target site in the HIV receptor gene. 
     
     
         3 . The method of  claim 1 , wherein a polynucleotide encoding the meganuclease is introduced into the cell. 
     
     
         4 . The method of  claim 1 , wherein the homing endonuclease is introduced into the cell ex vivo. 
     
     
         5 . The method of  claim 1  wherein the homing endonuclease is introduced into the cell in vivo. 
     
     
         6 . The method of  claim 3 , wherein the homing endonuclease is introduced into the cell ex vivo. 
     
     
         7 . The method of  claims 3 , wherein the homing endonuclease is introduced into the cell in vivo. 
     
     
         8 . The method of  claim 1 , wherein the HIV receptor is a CCR-5 receptor. 
     
     
         9 . The method of  claim 1 , wherein the HIV receptor is a CXCR4 receptor. 
     
     
         10 . The method of  claim 2  wherein the cell is selected from the group consisting of a stem cell, a T-cell, a macrophage, a dendritic cell and an antigen-presenting cell. 
     
     
         11 . The method of  claim 1 , wherein the homing endonuclease is selected from the group consisting of I-SceI, I-CeuI, PI-PspI, PI-Sce, I-SceIV, I-CsmI, I-PanI, I-SceII, I-PpoI, I-SceIII, I-CreI, I-TevI, I-TevII and I-TevIII. 
     
     
         12 . The method of  claim 9 , wherein the homing endonuclease is SceI or CreI. 
     
     
         13 . A method of treating or preventing HIV infection in a subject, the method comprising:
 inactivating an HIV receptor according to the method of  claim 1  in a cell isolated from the subject; and   introducing the cell into the subject.   
     
     
         14 . A method of treating or preventing HIV infection in a subject, the method comprising:
 introducing the polynucleotide encoding the meganuclease according to  claim 7  into a subject; and   inactivating an HIV receptor.