Genetic modification of cytokine inducible sh2-containing protein (cish) gene

Abstract

The present disclosure is in the field of genome engineering, particularly targeted genetic modification of a CISH gene.

1 .- 10 . (canceled) 
     
     
         11 . A zinc finger protein comprising 6 zinc finger domains each comprising a recognition helix region, wherein the zinc finger protein comprises the recognition helix regions of the proteins designated F1-F6 in the order F1 to F6, and wherein the recognition helix region comprises the proteins:
 F1 (SEQ ID NO: 2), F2 (SEQ ID NO: 3), F3 (SEQ ID NO: 4), F4 (SEQ ID NO: 5), F5 (SEQ ID NO: 6), and F6 (SEQ ID NO: 7);   F1 (SEQ ID NO: 8), F2 (SEQ ID NO: 9), F3 (SEQ ID NO: 10), F4 (SEQ ID NO: 11), F5 (SEQ ID NO: 9), and F6 (SEQ ID NO: 12);   F1 (SEQ ID NO: 13), F2 (SEQ ID NO: 14), F3 (SEQ ID NO: 10), F4 (SEQ ID NO: 15), F5 (SEQ ID NO: 16), and F6 (SEQ ID NO: 17);   F1 (SEQ ID NO: 18), F2 (SEQ ID NO: 19), F3 (SEQ ID NO: 20), F4 (SEQ ID NO: 21), F5 (SEQ ID NO: 18), and F6 (SEQ ID NO: 22);   F1 (SEQ ID NO: 23), F2 (SEQ ID NO: 24), F3 (SEQ ID NO: 25), F4 (SEQ ID NO: 26), F5 (SEQ ID NO: 27), and F6 (SEQ ID NO: 28);   F1 (SEQ ID NO: 29), F2 (SEQ ID NO: 30), F3 (SEQ ID NO: 16), F4 (SEQ ID NO: 31), F5 (SEQ ID NO: 32), and F6 (SEQ ID NO: 33);   F1 (SEQ ID NO: 34), F2 (SEQ ID NO: 35), F3 (SEQ ID NO: 32), F4 (SEQ ID NO: 36), F5 (SEQ ID NO: 37), and F6 (SEQ ID NO: 38); or   F1 (SEQ ID NO: 27), F2 (SEQ ID NO: 28), F3 (SEQ ID NO: 25), F4 (SEQ ID NO: 39), F5 (SEQ ID NO: 32), and F6 (SEQ ID NO: 12).   
     
     
         12 . A fusion protein comprising the zinc finger protein of  claim 11  and a wild-type or engineered cleavage domain or wild-type or engineered cleavage half-domain. 
     
     
         13 . A polynucleotide encoding one or more proteins of  claim 11 . 
     
     
         14 . An isolated cell comprising one or more proteins of  claim 12 . 
     
     
         15 . The isolated cell of  claim 14  or a cell descended therefrom, wherein the cell is selected from the group consisting of a T effector cell, a T regulatory cell and a hematopoietic stem cell. 
     
     
         16 . A kit comprising the protein of  claim 11 . 
     
     
         17 .- 19 . (canceled) 
     
     
         20 . A pharmaceutical composition comprising a zinc finger protein of  claim 11 . 
     
     
         21 . A pharmaceutical composition comprising a polynucleotide of  claim 13 . 
     
     
         22 . A pharmaceutical composition comprising an isolated cell of  claim 14 . 
     
     
         23 . A method of integrating a donor polynucleotide into an endogenous CISH gene comprising:
 (a) contacting one or more cells with a zinc finger nuclease to cleave the endogenous CISH gene, wherein the zinc finger nuclease comprises six zinc finger domains, wherein each zinc finger domain comprises a recognition helix region designated F1-F6 in the order F1 to F6, and wherein the recognition helix region comprises the proteins:   F1 (SEQ ID NO: 2), F2 (SEQ ID NO: 3), F3 (SEQ ID NO: 4), F4 (SEQ ID NO: 5), F5 (SEQ ID NO: 6), and F6 (SEQ ID NO: 7);   F1 (SEQ ID NO: 8), F2 (SEQ ID NO: 9), F3 (SEQ ID NO: 10), F4 (SEQ ID NO: 11), F5 (SEQ ID NO: 9), and F6 (SEQ ID NO: 12);   F1 (SEQ ID NO: 13), F2 (SEQ ID NO: 14), F3 (SEQ ID NO: 10), F4 (SEQ ID NO: 15), F5 (SEQ ID NO: 16), and F6 (SEQ ID NO: 17);   F1 (SEQ ID NO: 18), F2 (SEQ ID NO: 19), F3 (SEQ ID NO: 20), F4 (SEQ ID NO: 21), F5 (SEQ ID NO: 18), and F6 (SEQ ID NO: 22);   F1 (SEQ ID NO: 23), F2 (SEQ ID NO: 24), F3 (SEQ ID NO: 25), F4 (SEQ ID NO: 26), F5 (SEQ ID NO: 27), and F6 (SEQ ID NO: 28);   F1 (SEQ ID NO: 29), F2 (SEQ ID NO: 30), F3 (SEQ ID NO: 16), F4 (SEQ ID NO: 31), F5 (SEQ ID NO: 32), and F6 (SEQ ID NO: 33);   F1 (SEQ ID NO: 34), F2 (SEQ ID NO: 35), F3 (SEQ ID NO: 32), F4 (SEQ ID NO: 36), F5 (SEQ ID NO: 37), and F6 (SEQ ID NO: 38); or   F1 (SEQ ID NO: 27), F2 (SEQ ID NO: 28), F3 (SEQ ID NO: 25), F4 (SEQ ID NO: 39), F5 (SEQ ID NO: 32), and F6 (SEQ ID NO: 12); and   (b) introducing a donor polynucleotide into the one or more cells, such that the donor polynucleotide is integrated into the cleaved endogenous CISH gene.   
     
     
         24 . The method of  claim 23 , wherein the donor polynucleotide comprises a polynucleotide sequence encoding a Chimeric Antigen Receptor (CAR), an engineered or endogenous T cell receptor (TCR), an antibody-coupled T cell receptor (ACTR), or any combination thereof.