Journal
YEAST
Volume 33, Issue 10, Pages 549-557Publisher
WILEY
DOI: 10.1002/yea.3178
Keywords
Schizosaccharomyces pombe; fluoride channel; plasmid; CRISPR; Cas9
Categories
Funding
- Pilot Grant from the Yale Comprehensive Cancer Center
- American Cancer Society [IRG-58-012-57]
- NIH/NIGMS [R01GM115636]
Ask authors/readers for more resources
Fission yeast is a powerful model organism that has provided insights into important cellular processes thanks to the ease of its genome editing by homologous recombination. However, creation of strains with a large number of targeted mutations or containing plasmids has been challenging because only a very small number of selection markers is available in Schizosaccharomyces pombe. In this paper, we identify two fission yeast fluoride exporter channels (Fex1p and Fex2p) and describe the development of a new strategy using Fex1p as a selection marker for transformants in rich media supplemented with fluoride. To our knowledge this is the first positive selection marker identified in S. pombe that does not use auxotrophy or drug resistance and that can be used for plasmids transformation or genomic integration in rich media. We illustrate the application of our new marker by significantly accelerating the protocol for genome edition using CRISPR/Cas9 in S. pombe. Copyright (c) 2016 John Wiley & Sons, Ltd.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available