title: Development of a genome editing technique using the CRISPR/Cas9 system in the industrial filamentous fungus Aspergillus oryzae.
abstract: OBJECTIVE
  To develop a genome editing method using the CRISPR/Cas9 system in Aspergillus oryzae, the industrial filamentous fungus used in Japanese traditional fermentation and for the production of enzymes and heterologous proteins.
  
  
  RESULTS
  To develop the CRISPR/Cas9 system as a genome editing technique for A. oryzae, we constructed plasmids expressing the gene encoding Cas9 nuclease and single guide RNAs for the mutagenesis of target genes. We introduced these into an A. oryzae strain and obtained transformants containing mutations within each target gene that exhibited expected phenotypes. The mutational rates ranged from 10 to 20 %, and 1 bp deletions or insertions were the most commonly induced mutations.
  
  
  CONCLUSIONS
  We developed a functional and versatile genome editing method using the CRISPR/Cas9 system in A. oryzae. This technique will contribute to the use of efficient targeted mutagenesis in many A. oryzae industrial strains.
authors.affiliation: Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Katayama
authors.affiliation: Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Tanaka
authors.affiliation: Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
authors.forename: Tomoya
authors.initials: T
authors.lastname: Okabe
authors.affiliation: Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Nakamura
authors.affiliation: Department of Animal Resource Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
authors.forename: Wataru
authors.initials: W
authors.lastname: Fujii
authors.affiliation: Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Kitamoto
authors.affiliation: Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan. amarujun@mail.ecc.u-tokyo.ac.jp.
authors.forename: Jun-Ichi
authors.initials: J
authors.lastname: Maruyama
mesh_terms: D001236:Aspergillus oryzae / Q000235:genetics*; D064113:CRISPR-Cas Systems; D000072669:Gene Editing / Q000379:methods*; D016681:Genome, Fungal; D016296:Mutagenesis; D059645:Mutation Rate
keywords: Aspergillus oryzae; CRISPR/Cas9 system; Filamentous fungi; Genome editing