title: Targeted Mutagenesis in Bombyx mori Using TALENs.
abstract: Bombyx mori is a valuable model organism of high economic importance. Its genome sequence is available, as well as basic genetic and molecular genetic tools and markers. The introduction of genome editing methods based on engineered nucleases enables precise manipulations with genomic DNA, including targeted DNA deletions, insertions, or replacements in the genome allowing gene analysis and various applications. We describe here the use of TALENs which have a simple modular design of their DNA-binding domains, are easy to prepare and proved to be efficient in targeting of a wide range of cleavage sites. Our procedure often allows the production of individuals carrying homozygous mutations as early as in the G1 generation.
authors.affiliation: National Institute of Agrobiological Sciences, 1-2 Owashi, Tsukuba, Ibaraki, 305-8634, Japan.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Takasu
authors.affiliation: National Institute of Agrobiological Sciences, 1-2 Owashi, Tsukuba, Ibaraki, 305-8634, Japan. ttamura@affrc.go.jp.
authors.forename: Toshiki
authors.initials: T
authors.lastname: Tamura
authors.affiliation: Department of Biological Sciences, University of Rhode Island, 120 Flagg Road, Kingston, RI, 02881-0816, USA.
authors.forename: Marian
authors.initials: M
authors.lastname: Goldsmith
authors.affiliation: Biology Centre, Czech Academy of Sciences, Branisovska 31, 370 05, Ceske Budejovice, Czech Republic. zurovec@entu.cas.cz.
authors.forename: Michal
authors.initials: M
authors.lastname: Zurovec
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D012831:Bombyx / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics; D004720:Endonucleases / Q000737:chemistry / Q000235:genetics*; D018390:Gene Targeting / Q000379:methods*; D049750:Genome, Insect; D006801:Humans; D016296:Mutagenesis; D015534:Trans-Activators / Q000235:genetics*
chemical_list: D004268:DNA-Binding Proteins; D015534:Trans-Activators; D004720:Endonucleases
keywords: Engineered nucleases; Golden Gate assembly; NHEJ; Nonhomologous end joining; Silkworm; pBlue-TAL