title: ACandida albicansCRISPR system permits genetic engineering of essential genes and gene families.
abstract: Candida albicansis 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 aC. albicansCRISPR 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 ofCandidaand provides a new window into the biology of this pathogen.
authors.affiliation: Whitehead Institute for Biomedical Research, Cambridge, MA 02142, USA.
authors.forename: Valmik K
authors.initials: VK
authors.lastname: Vyas
authors.affiliation: Whitehead Institute for Biomedical Research, Cambridge, MA 02142, USA.
authors.forename: M Inmaculada
authors.initials: MI
authors.lastname: Barrasa
authors.affiliation: Whitehead Institute for Biomedical Research, Cambridge, MA 02142, USA ; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142, USA.
authors.forename: Gerald R
authors.initials: GR
authors.lastname: Fink