A Candida albicans CRISPR system permits genetic engineering of essential genes and gene families

Candida albicans is a pathogenic yeast that causes mucosal and systematic infections with high mortality. The absence of facile molecular genetics has been a major impediment to analysis of pathogenesis. The lack of meiosis coupled with the absence of plasmids makes genetic engineering cumbersome, e...

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Main Authors: Vyas, Valmik K., Barrasa, M. Inmaculada, Fink, Gerald R
Other Authors: Massachusetts Institute of Technology. Department of Biology
Format: Article
Language:en_US
Published: American Association for the Advancement of Science (AAAS) 2015
Online Access:http://hdl.handle.net/1721.1/96395
https://orcid.org/0000-0003-3704-2899
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author Vyas, Valmik K.
Barrasa, M. Inmaculada
Fink, Gerald R
author2 Massachusetts Institute of Technology. Department of Biology
author_facet Massachusetts Institute of Technology. Department of Biology
Vyas, Valmik K.
Barrasa, M. Inmaculada
Fink, Gerald R
author_sort Vyas, Valmik K.
collection MIT
description Candida albicans is a pathogenic yeast that causes mucosal and systematic infections with high mortality. The absence of facile molecular genetics has been a major impediment to analysis of pathogenesis. The lack of meiosis coupled with the absence of plasmids makes genetic engineering cumbersome, especially for essential functions and gene families. We describe a C. albicans CRISPR system that overcomes many of the obstacles to genetic engineering in this organism. The high frequency with which CRISPR-induced mutations can be directed to target genes enables easy isolation of homozygous gene knockouts, even without selection. Moreover, the system permits the creation of strains with mutations in multiple genes, gene families, and genes that encode essential functions. This CRISPR system is also effective in a fresh clinical isolate of undetermined ploidy. Our method transforms the ability to manipulate the genome of Candida and provides a new window into the biology of this pathogen.
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spelling mit-1721.1/963952022-09-27T14:59:30Z A Candida albicans CRISPR system permits genetic engineering of essential genes and gene families Vyas, Valmik K. Barrasa, M. Inmaculada Fink, Gerald R Massachusetts Institute of Technology. Department of Biology Whitehead Institute for Biomedical Research Fink, Gerald R. Candida albicans is a pathogenic yeast that causes mucosal and systematic infections with high mortality. The absence of facile molecular genetics has been a major impediment to analysis of pathogenesis. The lack of meiosis coupled with the absence of plasmids makes genetic engineering cumbersome, especially for essential functions and gene families. We describe a C. albicans CRISPR system that overcomes many of the obstacles to genetic engineering in this organism. The high frequency with which CRISPR-induced mutations can be directed to target genes enables easy isolation of homozygous gene knockouts, even without selection. Moreover, the system permits the creation of strains with mutations in multiple genes, gene families, and genes that encode essential functions. This CRISPR system is also effective in a fresh clinical isolate of undetermined ploidy. Our method transforms the ability to manipulate the genome of Candida and provides a new window into the biology of this pathogen. National Institutes of Health (U.S.) (GM035010) 2015-04-07T16:50:04Z 2015-04-07T16:50:04Z 2015-04 2015-02 Article http://purl.org/eprint/type/JournalArticle 2375-2548 http://hdl.handle.net/1721.1/96395 Vyas, Valmik K., M. Inmaculada Barrasa, and Gerald R. Fink. “A Candida Albicans CRISPR System Permits Genetic Engineering of Essential Genes and Gene Families.” Science Advances 1, no. 3 (April 1, 2015): e1500248–e1500248. https://orcid.org/0000-0003-3704-2899 en_US http://dx.doi.org/10.1126/sciadv.1500248 Science Advances Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc/4.0/ application/pdf American Association for the Advancement of Science (AAAS) Science Advances
spellingShingle Vyas, Valmik K.
Barrasa, M. Inmaculada
Fink, Gerald R
A Candida albicans CRISPR system permits genetic engineering of essential genes and gene families
title A Candida albicans CRISPR system permits genetic engineering of essential genes and gene families
title_full A Candida albicans CRISPR system permits genetic engineering of essential genes and gene families
title_fullStr A Candida albicans CRISPR system permits genetic engineering of essential genes and gene families
title_full_unstemmed A Candida albicans CRISPR system permits genetic engineering of essential genes and gene families
title_short A Candida albicans CRISPR system permits genetic engineering of essential genes and gene families
title_sort candida albicans crispr system permits genetic engineering of essential genes and gene families
url http://hdl.handle.net/1721.1/96395
https://orcid.org/0000-0003-3704-2899
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