title: Site-Specific Integration of Exogenous Genes Using Genome Editing Technologies in Zebrafish.
abstract: The zebrafish (Danio rerio) is an ideal vertebrate model to investigate the developmental molecular mechanism of organogenesis and regeneration. Recent innovation in genome editing technologies, such as zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9) system, have allowed researchers to generate diverse genomic modifications in whole animals and in cultured cells. The CRISPR/Cas9 and TALEN techniques frequently induce DNA double-strand breaks (DSBs) at the targeted gene, resulting in frameshift-mediated gene disruption. As a useful application of genome editing technology, several groups have recently reported efficient site-specific integration of exogenous genes into targeted genomic loci. In this review, we provide an overview of TALEN- and CRISPR/Cas9-mediated site-specific integration of exogenous genes in zebrafish.
authors.affiliation: Laboratory for Developmental Biology, Center for Medical Education and Sciences, Graduate School of Medical Science, University of Yamanashi, 1110 Shimokato, Chuo, Yamanashi 409-3898, Japan. akawahara@yamanashi.ac.jp.
authors.forename: Atsuo
authors.initials: A
authors.lastname: Kawahara
authors.affiliation: Laboratory for Developmental Gene Regulation, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan. y.hisano@riken.jp.
authors.forename: Yu
authors.initials: Y
authors.lastname: Hisano
authors.affiliation: Laboratory for Developmental Biology, Center for Medical Education and Sciences, Graduate School of Medical Science, University of Yamanashi, 1110 Shimokato, Chuo, Yamanashi 409-3898, Japan. satoshio-as@yamanashi.ac.jp.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Ota
authors.affiliation: Laboratory for Developmental Biology, Center for Medical Education and Sciences, Graduate School of Medical Science, University of Yamanashi, 1110 Shimokato, Chuo, Yamanashi 409-3898, Japan. ktaimatsu@yamanashi.ac.jp.
authors.forename: Kiyohito
authors.initials: K
authors.lastname: Taimatsu
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D000072669:Gene Editing / Q000379:methods*; D055879:Gene Knock-In Techniques / Q000379:methods*; D011995:Recombination, Genetic; D015027:Zebrafish / Q000235:genetics*
keywords: CRISPR/Cas9 system; MMEJ; genome editing; knock-in; zebrafish