title: TALEN construction via "Unit Assembly" method and targeted genome modifications in zebrafish.
abstract: Transcription activator-like effector nucleases (TALENs) are engineered endonucleases composed of a customized transcription activator-like effector (TALE) DNA-binding domain and a FokI DNA cleavage domain. TALENs induce DNA double-strand breaks (DSBs) at their target sites on the chromosome and have been successfully used for genome engineering in many species and cultured cells. Zebrafish is a very popular model organism in both basic and clinical research. Here, we describe the details of construction of customized TALENs using the "Unit Assembly" (UA) method, as well as three applications of zebrafish genome manipulations using TALENs: gene knock-out, large chromosome deletion, and gene knock-in by homologous recombination.
authors.affiliation: Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing 100871, PR China.
authors.forename: Peng
authors.initials: P
authors.lastname: Huang
authors.affiliation: Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing 100871, PR China.
authors.forename: An
authors.initials: A
authors.lastname: Xiao
authors.affiliation: Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing 100871, PR China.
authors.forename: Xiangjun
authors.initials: X
authors.lastname: Tong
authors.affiliation: Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing 100871, PR China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Zu
authors.affiliation: Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing 100871, PR China.
authors.forename: Zhanxiang
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing 100871, PR China. Electronic address: bzhang@pku.edu.cn.
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
mesh_terms: D000818:Animals; D002872:Chromosome Deletion; D003851:Deoxyribonucleases / Q000737:chemistry* / Q000235:genetics; D055879:Gene Knock-In Techniques; D055786:Gene Knockout Techniques; D005818:Genetic Engineering / Q000379:methods*; D018095:Germ-Line Mutation; D059765:Homologous Recombination; D015202:Protein Engineering / Q000379:methods; D015027:Zebrafish / Q000235:genetics
chemical_list: D003851:Deoxyribonucleases
keywords: Chromosomal deletion; Gene targeting; Genome manipulation; Homologous recombination; TALEN; Zebrafish