Creating knockout and knockin rodents using engineered endonucleases via direct embryo injection

Abstract

Genetically engineered rodents have been generated worldwide for biomedical research. Recently, gene targeting techniques have been developed by using engineered endonucleases such as zinc-fi nger nucleases (ZFN), transcription activator-like effector nucleases (TALEN) and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9. These endonucleases are useful for simple and rapid production of gene knockout/knockin animals without using embryonic stem (ES) cells. This chapter introduces the latest protocols for producing genetically modifi ed rodents using ZFN, TALEN, and CRISPR/Cas9.