Research ArticleMOLECULAR GENETICS

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

Science Advances  03 Apr 2015:
Vol. 1, no. 3, e1500248
DOI: 10.1126/sciadv.1500248

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Abstract

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.

Keywords
  • Candida albicans
  • CRISPR
  • Cas9
  • genetics
  • mutagenesis
  • fungal pathogenesis
  • CaCas9
  • CTG clade

This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license, which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited.

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