A CRISPR–Cas9-based gene drive platform for genetic interaction analysis in Candida albicans
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nihms906721.pdf
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Author(s)
Shapiro, Rebecca S.
Chavez, Alejandro
Porter, Caroline B. M.
Hamblin, Meagan
Kaas, Christian S.
DiCarlo, James E.
Zeng, Guisheng
Xu, Xiaoli
Revtovich, Alexey V.
Kirienko, Natalia V.
Date Issued
October 2017
Journal
Nature Microbiology
Publisher
Nature Publishing Group
Citation
Shapiro, Rebecca S. et al. “A CRISPR–Cas9-Based Gene Drive Platform for Genetic Interaction Analysis in Candida Albicans.” Nature Microbiology 3, 1 (October 2017): 73–82 © 2017 The Author(s)
Version
Author's final manuscript
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.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Institute for Medical Engineering & Science
Massachusetts Institute of Technology. Synthetic Biology Center
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Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
https://doi.org/10.1038/S41564-017-0043-0
