title: 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-finger 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 modified rodents using ZFN, TALEN, and CRISPR/Cas9.
authors.affiliation: Institute of Laboratory Animals, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto, 606-8501, Japan, tkaneko@anim.med.kyoto-u.ac.jp.
authors.forename: Takehito
authors.initials: T
authors.lastname: Kaneko
authors.forename: Tomoji
authors.initials: T
authors.lastname: Mashimo
mesh_terms: D000818:Animals; D004624:Embryo Transfer / Q000379:methods; D004622:Embryo, Mammalian; D004720:Endonucleases / Q000235:genetics* / Q000378:metabolism*; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods; D055879:Gene Knock-In Techniques; D055786:Gene Knockout Techniques; D005818:Genetic Engineering / Q000379:methods*; D060005:Genotyping Techniques; D008297:Male; D051379:Mice; D008845:Microinjections / Q000379:methods*; D012333:RNA, Messenger / Q000008:administration & dosage / Q000235:genetics; D051381:Rats; D012377:Rodentia
chemical_list: D012333:RNA, Messenger; D004720:Endonucleases