title: Targeted Mutagenesis in Rice Using TALENs and the CRISPR/Cas9 System.
abstract: Sequence-specific nucleases (SSNs), such as zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 nuclease (Cas9) system, are powerful tools for understanding gene function and for developing novel traits in plants. In plant species for which transformation and regeneration systems using protoplasts are not yet established, direct delivery to nuclei of SSNs either in the form of RNA or protein is difficult. Thus, Agrobacterium-mediated transformation of SSN expression constructs in cultured cells is a practical means of delivering targeted mutagenesis in some plant species including rice. Because targeted mutagenesis occurs stochastically in transgenic cells and SSN-mediated targeted mutagenesis often leads to no selectable phenotype, identification of highly mutated cell lines is a critical step in obtaining regenerated plants with desired mutations.
authors.affiliation: Plant Genome Engineering Research Unit, National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Endo
authors.affiliation: Plant Genome Engineering Research Unit, National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan.
authors.forename: Ayako
authors.initials: A
authors.lastname: Nishizawa-Yokoi
authors.affiliation: Plant Genome Engineering Research Unit, National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan. stoki@affrc.go.jp.|Kihara Institute for Biological Research, Yokohama City University, 641-12 Maioka-cho, Yokohama, 244-0813, Japan. stoki@affrc.go.jp.
authors.forename: Seiichi
authors.initials: S
authors.lastname: Toki
mesh_terms: D060054:Agrobacterium / Q000235:genetics; D064113:CRISPR-Cas Systems; D005822:Genetic Vectors; D020180:Heteroduplex Analysis; D016296:Mutagenesis; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D059645:Mutation Rate; D012275:Oryza / Q000235:genetics*; D030821:Plants, Genetically Modified; D000069896:Transcription Activator-Like Effector Nucleases / Q000235:genetics*
chemical_list: D000069896:Transcription Activator-Like Effector Nucleases
keywords: CRISPR/Cas9; DNA double-strand breaks; Sequence-specific nuclease; TALENs; Targeted mutagenesis