title: Tailor-made TALEN system for highly efficient targeted gene replacement in the rice blast fungus.
abstract: Genetic manipulation is key to unraveling gene functions and creating genetically modified strains of microbial organisms. Recently, engineered nucleases that can generate DNA double-strand breaks (DSBs) at a specific site in the desired locus within genome are utilized in a rapidly developing genome editing technology via DSBs repair. However, the use of engineered nucleases in filamentous fungi has not been validated. In this study, we demonstrated that tailor-made transcriptional activator-like effector nucleases (TALENs) system, Platinum-Fungal TALENs (PtFg TALENs), could improve the efficiency of homologous recombination-mediated targeted gene replacement by up to 100% in the rice blast fungus Pyricularia oryzae. This high-efficiency PtFg TALEN has great potential for basic and applied biological applications in filamentous fungi.
authors: Graduate School of Agriculture, Meiji University, 1-1-1 Higashi-Mita, Tama-ku, Kawasaki, Kanagawa, 214-8571, Japan. arazoet@gmail.com.
authors: Takayuki
authors: T
authors: Arazoe
authors: Graduate School of Agriculture, Meiji University, 1-1-1 Higashi-Mita, Tama-ku, Kawasaki, Kanagawa, 214-8571, Japan.
authors: Tetsuo
authors: T
authors: Ogawa
authors: Graduate School of Agriculture, Meiji University, 1-1-1 Higashi-Mita, Tama-ku, Kawasaki, Kanagawa, 214-8571, Japan.
authors: Kennosuke
authors: K
authors: Miyoshi
authors: Graduate School of Agriculture, Meiji University, 1-1-1 Higashi-Mita, Tama-ku, Kawasaki, Kanagawa, 214-8571, Japan.
authors: Tohru
authors: T
authors: Yamato
authors: Graduate School of Agriculture, Meiji University, 1-1-1 Higashi-Mita, Tama-ku, Kawasaki, Kanagawa, 214-8571, Japan.
authors: Shuichi
authors: S
authors: Ohsato
authors: Graduate School of Science, Hiroshima University, Higashi-Hiroshima, Japan.
authors: Tetsushi
authors: T
authors: Sakuma
authors: Graduate School of Science, Hiroshima University, Higashi-Hiroshima, Japan.
authors: Takashi
authors: T
authors: Yamamoto
authors: Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Tokyo, Japan.
authors: Tsutomu
authors: T
authors: Arie
authors: Graduate School of Agriculture, Meiji University, 1-1-1 Higashi-Mita, Tama-ku, Kawasaki, Kanagawa, 214-8571, Japan.
authors: Shigeru
authors: S
authors: Kuwata
mesh_terms: D018390:Gene Targeting / Q000379:methods*; D005800:Genes, Fungal; D005827:Genetics, Microbial / Q000379:methods*; D059765:Homologous Recombination; D020082:Magnaporthe / Q000235:genetics*; D008967:Molecular Biology / Q000379:methods*; D012275:Oryza / Q000382:microbiology; D010935:Plant Diseases / Q000382:microbiology
keywords: TALEN; blast fungus; engineered nuclease; filamentous fungi; gene targeting; genome editing; rice