Implementation of the CRISPR-Cas9 system in fission yeast

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 RNA (rrk1) and a ribozyme to produce the ta...

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Detalles Bibliográficos
Autores principales: Jacobs, Jake Z., Ciccaglione, Keith M., Tournier, Vincent, Zaratiegui, Mikel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2014
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4215166/
https://www.ncbi.nlm.nih.gov/pubmed/25352017
http://dx.doi.org/10.1038/ncomms6344
Descripción
Sumario: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 RNA (rrk1) and a ribozyme to produce the targeting guide RNA. Tobether with constitutive expressio nof 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.

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