A CRISPR–Cas9-based gene drive platform for genetic interaction analysis in Candida albicans
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 mo...
| Main Authors: | , , , , , , , , , , , , |
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| Other Authors: | |
| Format: | Article |
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Nature Publishing Group
2018
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| Online Access: | http://hdl.handle.net/1721.1/117583 |
| _version_ | 1826203274007543808 |
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| author | 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. Wang, Yue Church, George M. Collins, James J. |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering 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. Wang, Yue Church, George M. Collins, James J. |
| author_sort | Shapiro, Rebecca S. |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T12:34:02Z |
| format | Article |
| id | mit-1721.1/117583 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T12:34:02Z |
| publishDate | 2018 |
| publisher | Nature Publishing Group |
| record_format | dspace |
| spelling | mit-1721.1/1175832024-06-27T14:37:08Z A CRISPR–Cas9-based gene drive platform for genetic interaction analysis in Candida albicans 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. Wang, Yue Church, George M. Collins, James J. Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Institute for Medical Engineering & Science Massachusetts Institute of Technology. Synthetic Biology Center 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. National Human Genome Research Institute (U.S.) (Grant RM1HG008525) 2018-08-28T15:42:50Z 2018-08-28T15:42:50Z 2017-10 2017-06 2018-08-27T18:19:02Z Article http://purl.org/eprint/type/JournalArticle 2058-5276 http://hdl.handle.net/1721.1/117583 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) http://dx.doi.org/10.1038/S41564-017-0043-0 Nature Microbiology 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. application/pdf Nature Publishing Group PMC |
| spellingShingle | 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. Wang, Yue Church, George M. Collins, James J. 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 |
| title_full | A CRISPR–Cas9-based gene drive platform for genetic interaction analysis in Candida albicans |
| title_fullStr | A CRISPR–Cas9-based gene drive platform for genetic interaction analysis in Candida albicans |
| title_full_unstemmed | A CRISPR–Cas9-based gene drive platform for genetic interaction analysis in Candida albicans |
| title_short | A CRISPR–Cas9-based gene drive platform for genetic interaction analysis in Candida albicans |
| title_sort | crispr cas9 based gene drive platform for genetic interaction analysis in candida albicans |
| url | http://hdl.handle.net/1721.1/117583 |
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