Unexpected insertion of carrier DNA sequences into the fission yeast genome during CRISPR–Cas9 mediated gene deletion View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2019-12

AUTHORS

Sophie Longmuir, Nabihah Akhtar, Stuart A. MacNeill

ABSTRACT

OBJECTIVES: The fission yeast Schizosaccharomyces pombe is predicted to encode ~ 200 proteins of < 100 amino acids, including a number of previously uncharacterised proteins that are found conserved in related Schizosaccharomyces species only. To begin an investigation of the function of four of these so-called microproteins (designated Smp1-Smp4), CRISPR-Cas9 genome editing technology was used to delete the corresponding genes in haploid fission yeast cells. RESULTS: None of the four microprotein-encoding genes was essential for viability, meiosis or sporulation, and the deletion cells were no more sensitive to a range of cell stressors than wild-type, leaving the function of the proteins unresolved. During CRISPR-Cas9 editing however, a number of strains were isolated in which additional sequences were inserted into the target loci at the Cas9 cut site. Sequencing of the inserts revealed these to be derived from the chum salmon Oncorhynchus keta, the source of the carrier DNA used in the S. pombe transformation. More... »

PAGES

191

References to SciGraph publications

  • 2018-05-15. PomBase: The Scientific Resource for Fission Yeast in EUKARYOTIC GENOMIC DATABASES
  • 2014-12. Implementation of the CRISPR-Cas9 system in fission yeast in NATURE COMMUNICATIONS
  • Journal

    TITLE

    BMC Research Notes

    ISSUE

    1

    VOLUME

    12

    Author Affiliations

    Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1186/s13104-019-4228-x

    DOI

    http://dx.doi.org/10.1186/s13104-019-4228-x

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1113099420

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/30925937


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