title: Implementation of the CRISPR-Cas9 system in fission yeast.
abstract: Application of the CRISPR-Cas9 genome editing system in the model organism Schizosaccharomyces pombe has been hampered by the lack of constructs to express RNA of arbitrary sequence. Here we present expression constructs that use the promoter/leader RNA of K RNA (rrk1) and a ribozyme to produce the targeting guide RNA. Together with constitutive expression of Cas9, this system achieves selection-free specific mutagenesis with efficiencies approaching 100%. The rrk1 CRISPR-Cas9 method enables rapid and efficient genome manipulation and unlocks the CRISPR toolset for use in fission yeast.
authors.affiliation: Department of Molecular Biology and Biochemistry, Rutgers, the State University of New Jersey, Piscataway, New Jersey 08854, USA.
authors.forename: Jake Z
authors.initials: JZ
authors.lastname: Jacobs
authors.affiliation: Department of Molecular Biology and Biochemistry, Rutgers, the State University of New Jersey, Piscataway, New Jersey 08854, USA.
authors.forename: Keith M
authors.initials: KM
authors.lastname: Ciccaglione
authors.affiliation: Department of Molecular Biology and Biochemistry, Rutgers, the State University of New Jersey, Piscataway, New Jersey 08854, USA.
authors.forename: Vincent
authors.initials: V
authors.lastname: Tournier
authors.affiliation: Department of Molecular Biology and Biochemistry, Rutgers, the State University of New Jersey, Piscataway, New Jersey 08854, USA.
authors.forename: Mikel
authors.initials: M
authors.lastname: Zaratiegui
mesh_terms: D064130:CRISPR-Associated Proteins / Q000378:metabolism*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D018604:Epitope Mapping; D015966:Gene Expression Regulation, Fungal; D005822:Genetic Vectors / Q000378:metabolism; D016296:Mutagenesis / Q000235:genetics; D009154:Mutation / Q000235:genetics; D011401:Promoter Regions, Genetic; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000235:genetics*
chemical_list: D064130:CRISPR-Associated Proteins; D012331:RNA, Fungal; D000094704:RNA, Guide, CRISPR-Cas Systems