Efficient non-meiotic allele introgression in livestock

Abstract

Methods, uses, and animals for introgression of alleles between animals, including SNPs. One embodiment involves introducing a targeted targeting endonuclease system and a HDR template into a cell with a mismatch in the binding of the targeting endonuclease and the targeted site.

1 . A method of modifying a bovine cell, the method comprising:
 contacting the bovine cell with a targeted endonuclease that targets and cuts a gene encoding the prolactin receptor;   contacting the bovine cell with a homology-dependent repair template such that the template integrates into the genome of the bovine cell to encode a truncated prolactin receptor protein.   
     
     
         2 . The method of  claim 1 , wherein the truncated prolactin receptor protein is 461 amino acids in length. 
     
     
         3 . The method of  claim 1 , wherein the targeted endonuclease is selected from a zinc finger nuclease, a TAL effector nuclease (TALEN), and a CRISPR/Cas 9 nuclease. 
     
     
         4 . The method of  claim 3 , wherein the targeted endonuclease is a TAL effector nuclease (TALEN). 
     
     
         5 . The method of  claim 4 , wherein the TALEN has zero mismatches to a targeted region of the gene encoding the prolactin receptor. 
     
     
         6 . The method of  claim 1 , wherein contacting the bovine cell with a targeted endonuclease comprises expressing exogenous mRNA encoding a TAL effector nuclease (TALEN).