Content area
Full Text
About the Authors:
Yun Liu
Contributed equally to this work with: Yun Liu, Daji Luo
Affiliations School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong, China, School of Biomedical Sciences Core Laboratory, The Chinese University of Hong Kong Shenzhen Research Institute, Shenzhen, China
Daji Luo
Contributed equally to this work with: Yun Liu, Daji Luo
Affiliations State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China, Department of Genetics, School of Basic Medical Sciences, Wuhan University, Wuhan, China
Hui Zhao
Affiliations School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong, China, School of Biomedical Sciences Core Laboratory, The Chinese University of Hong Kong Shenzhen Research Institute, Shenzhen, China
Zuoyan Zhu
Affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China
Wei Hu
* E-mail: [email protected] (WH); [email protected] (CC)
Affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China
Christopher H. K. Cheng
* E-mail: [email protected] (WH); [email protected] (CC)
Affiliations School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong, China, School of Biomedical Sciences Core Laboratory, The Chinese University of Hong Kong Shenzhen Research Institute, Shenzhen, China
Introduction
TALENs are artificial nucleases that consist of the DNA binding domain from transcription activator like effectors (TALE) and a catalytic domain from FokI nuclease [1]–[2]. TALEs bind to DNA through the repeat domain, with one TALE repeat recognizing one DNA base. The base specificity is determined by the 12th and 13th amino acids called repeat-variable di-residues (RVDs) in each TALE repeat, with RVDs NI, NG, HD and NN recognizing adenine (A), thymine (T), cytosine (C) and guanine (G), respectively [3]–[4]. These RVDs-DNA pairings bring the FokI nuclease to a predetermined genomic locus to create DNA double-strand breaks (DSB). Repair of the DSB through the error-prone non-homologous end-joining pathway leads to small indels at the break site, thus enabling targeted gene disruption. So far, TALENs have been employed to disrupt specific genomic loci in yeast [5], worms [1], [6]–[7], plants [8], zebrafish [9]–[13], medaka [14], rat [15], Xenopus [16], pig [17] as well as in cell lines [18]–[20] and human stem cells [21].
The indel-mutations...