Unexpected insertion of carrier DNA sequences into the fission yeast genome during CRISPR–Cas9 mediated gene deletion
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.
Citation
Longmuir , S , Akhtar , N & MacNeill , S A 2019 , ' Unexpected insertion of carrier DNA sequences into the fission yeast genome during CRISPR–Cas9 mediated gene deletion ' , BMC Research Notes , vol. 12 , 191 . https://doi.org/10.1186/s13104-019-4228-x
Publication
BMC Research Notes
Status
Peer reviewed
ISSN
1756-0500Type
Journal article
Rights
© The Author(s) 2019. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licen ses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Description
Funding: This work was funded by the School of Biology, University of St Andrews, and by the Nuffield Foundation.Collections
Items in the St Andrews Research Repository are protected by copyright, with all rights reserved, unless otherwise indicated.