title: Seamless Genome Editing in Rice via Gene Targeting and Precise Marker Elimination.
abstract: Positive-negative selection using hygromycin phosphotransferase (hpt) and diphtheria toxin A-fragment (DT-A) as positive and negative selection markers, respectively, allows enrichment of cells harboring target genes modified via gene targeting (GT). We have developed a successful GT system employing positive-negative selection and subsequent precise marker excision via the piggyBac transposon derived from the cabbage looper moth to introduce desired modifications into target genes in the rice genome. This approach could be applied to the precision genome editing of almost all endogenous genes throughout the genome, at least in rice.
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.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Saika
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: D004251:DNA Transposable Elements; D004167:Diphtheria Toxin / Q000235:genetics; D000072669:Gene Editing / Q000379:methods*; D018390:Gene Targeting / Q000379:methods*; D005819:Genetic Markers; D005822:Genetic Vectors; D018745:Genome, Plant; D012275:Oryza / Q000235:genetics*; D010446:Peptide Fragments / Q000235:genetics; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction
chemical_list: D004251:DNA Transposable Elements; D004167:Diphtheria Toxin; D005819:Genetic Markers; D010446:Peptide Fragments; C029795:diphtheria toxin fragment A; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase
keywords: Gene targeting; Homologous recombination; Marker excision; Positive–negative selection; piggyBac transposon