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Nat Commun. 2014 Oct 29;5:5344. doi: 10.1038/ncomms6344.

Implementation of the CRISPR-Cas9 system in fission yeast.

Author information

1
Department of Molecular Biology and Biochemistry, Rutgers, the State University of New Jersey, Piscataway, New Jersey 08854, USA.

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.

PMID:
25352017
PMCID:
PMC4215166
DOI:
10.1038/ncomms6344
[Indexed for MEDLINE]
Free PMC Article

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