title: Efficient production of a gene mutant cell line through integrating TALENs and high-throughput cell cloning.
abstract: Transcription activator-like effectors (TALEs) are becoming powerful DNA-targeting tools in a variety of mammalian cells and model organisms. However, generating a stable cell line with specific gene mutations in a simple and rapid manner remains a challenging task. Here, we report a new method to efficiently produce monoclonal cells using integrated TALE nuclease technology and a series of high-throughput cell cloning approaches. Following this method, we obtained three mTOR mutant 293T cell lines within 2 months, which included one homozygous mutant line.
authors.affiliation: Department of Biomedical Engineering, College of Engineering, Peking University, Beijing, China.
authors.forename: Changhong
authors.initials: C
authors.lastname: Sun
authors.affiliation: Department of Biomedical Engineering, College of Engineering, Peking University, Beijing, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Fan
authors.affiliation: Suzhou Genoarray Co., Ltd., Suzhou, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Biomedical Engineering, College of Engineering, Peking University, Beijing, China.
authors.forename: Gancheng
authors.initials: G
authors.lastname: Wang
authors.affiliation: Department of Biomedical Engineering, College of Engineering, Peking University, Beijing, China.
authors.forename: Hanshuo
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Biomedical Engineering, College of Engineering, Peking University, Beijing, China State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Molecular Medicine, Peking University, Beijing, China jzxi@pku.edu.cn shzzhshuo@sina.com.
authors.forename: Jianzhong Jeff
authors.initials: JJ
authors.lastname: Xi
mesh_terms: D002460:Cell Line; D019976:Cloning, Organism / Q000379:methods*; D003584:Cytological Techniques / Q000379:methods*; D018390:Gene Targeting / Q000379:methods*; D005818:Genetic Engineering / Q000379:methods*; D057166:High-Throughput Screening Assays; D006801:Humans
keywords: TALEN; gene mutant; high-throughput; stable cell line