Method for CRISPR/Cas9 Mutagenesis in Candida albicans

Neta Dean; Henry Ng

doi:10.21769/BioProtoc.2814

Method for CRISPR/Cas9 Mutagenesis in Candida albicans

Bio Protoc. 2018 Apr 20;8(8):e2814. doi: 10.21769/BioProtoc.2814.

Authors

Neta Dean¹, Henry Ng²

Affiliations

¹ Department of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, New York, USA.
² Department of Molecular Biology, University of California, San Francisco, CA, USA.

Abstract

Candida albicans is the most prevalent and important human fungal pathogen. The advent of CRISPR as a means of gene editing has greatly facilitated genetic analysis in C. albicans. Here, we describe a detailed step-by-step procedure to construct and analyze C. albicans deletion mutants. This protocol uses plasmids that allow simple ligation of synthetic duplex 23mer guide oligodeoxynucleotides for high copy gRNA expression in C. albicans strains that express codon-optimized Cas9. This protocol allows isolation and characterization of deletion strains within nine days.

Keywords: CRISPR; Candida albicans; Cas9; Fungal genetics; Yeast; gRNA.