A CRISPR–Cas9-based gene drive platform for genetic interaction analysis in Candida albicans

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A CRISPR–Cas9-based gene drive platform for genetic interaction analysis in Candida albicans
Title:
A CRISPR–Cas9-based gene drive platform for genetic interaction analysis in Candida albicans
Journal Title:
Nature Microbiology
Publication Date:
23 October 2017
Citation:
Shapiro, R. S., Chavez, A., Porter, C. B. M., Hamblin, M., Kaas, C. S., DiCarlo, J. E., … Collins, J. J. (2017). A CRISPR–Cas9-based gene drive platform for genetic interaction analysis in Candida albicans. Nature Microbiology, 3(1), 73–82. doi:10.1038/s41564-017-0043-0
Abstract:
Candida albicans is the leading cause of fungal infections; yet, complex genetic interaction analysis remains cumbersome in this diploid pathogen. Here, we developed a CRISPR–Cas9-based ‘gene drive array’ platform to facilitate efficient genetic analysis in C. albicans. In our system, a modified DNA donor molecule acts as a selfish genetic element, replaces the targeted site and propagates to replace additional wild-type loci. Using mating-competent C. albicans haploids, each carrying a different gene drive disabling a gene of interest, we are able to create diploid strains that are homozygous double-deletion mutants. We generate double-gene deletion libraries to demonstrate this technology, targeting antifungal efflux and biofilm adhesion factors. We screen these libraries to identify virulence regulators and determine how genetic networks shift under diverse conditions. This platform transforms our ability to perform genetic interaction analysis in C. albicans and is readily extended to other fungal pathogens.
License type:
Publisher Copyright
Funding Info:
We thank G. Fink, J. Berman, M. Hickman, V. Vyas and A. Baryshnikova for helpful discussions. We also thank V. Vyas, J. Köhler and L. Cowen for strains. This work was supported by the Paul G. Allen Frontiers Group, a Banting postdoctoral fellowship from the Canadian Institutes of Health Research, National Cancer Institute grantno. 5T32CA009216-34, US National Institutes of Health National Human Genome Research Institute grant no. RM1 HG008525 and the Wyss Institute for Biologically Inspired Engineering.This work was supported by the IMCB core budget
Description:
ISSN:
2058-5276
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