title: Mutagenesis in Xenopus and Zebrafish using TALENs.
abstract: Transcription activator-like effector nucleases (TALENs) have been proven to be effective for gene specific targeting across species. Here we describe the validated protocol for TALEN assembly as well as methods for generating gene knockout animals of Xenopus and zebrafish. This protocol covers selection of TALEN targeting sites, TALEN assembly with a modified Golden Gate method, injection of TALEN mRNAs into Xenopus and zebrafish embryos as well as the detection of somatic and germ-line transmitted mutations. Finally, the establishment of knockout Xenopus and zebrafish lines is also described. This protocol will facilitate broader applications of TALENs in developmental biology.
authors.affiliation: School of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China.|School of Biomedical Sciences Core Laboratory, Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, 518057, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Liu
authors.affiliation: School of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China. zhaohui@cuhk.edu.hk.|School of Biomedical Sciences Core Laboratory, Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, 518057, China. zhaohui@cuhk.edu.hk.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhao
authors.affiliation: School of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China. chkcheng@cuhk.edu.hk.|School of Biomedical Sciences Core Laboratory, Shenzhen Research Institute, The Chinese University of Hong Kong, Shenzhen, 518057, China. chkcheng@cuhk.edu.hk.
authors.forename: Christopher H K
authors.initials: CH
authors.lastname: Cheng
mesh_terms: D000818:Animals; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism; D055786:Gene Knockout Techniques / Q000379:methods*; D005818:Genetic Engineering / Q000379:methods*; D023281:Genomics; D006801:Humans; D016296:Mutagenesis / Q000235:genetics; D015534:Trans-Activators / Q000235:genetics*; D014981:Xenopus / Q000235:genetics*; D015027:Zebrafish / Q000235:genetics
chemical_list: D015534:Trans-Activators; D004720:Endonucleases
keywords: Gene disruption; Gene knockout; Mutagenesis; TALENs; Xenopus; Zebrafish