Many knock-out/knock-in mouse and rat strains have been produced by genome editing techniques using engineered endonucleases, including zinc finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN), or clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9. Microinjection of engineered endonucleases into pronuclear-stage embryos is required to produce genome-edited rodents and the development of easy, rapid, and high-efficiency methods that do not require special skills such as microinjection is needed. This chapter presents a new technique called Technique for Animal Knockout system by Electroporation (TAKE), which produces genome-edited rodents by direct introduction of engineered endonucleases into intact embryos using electroporation.
Keywords: CRISPR/Cas9; Cas9 protein; Electroporation; Intact embryos; Knock-in; Knock-out; Mouse; Rat; TALEN; ZFN.