Downregulation of Essential Genes in the Fungal Pathogen Candida auris

The recent global emergence of the fungal pathogen Candida auris has caused significant concern given that this pathogen often exhibits resistance to multiple antifungal drug classes. In order to effectively combat C. auris infections, there is a dire need to expand our current antifungal arsenal. Essential proteins often serve as targets for antimicrobial compounds, and thus being able to study essential genes in a pathogen of interest is a critical first step in drug development. To identify and characterize essential genes in microorganisms, researchers must be able to manipulate microbial genomes using a variety of molecular biology techniques. Given the haploid genome of C. auris, genetic alterations have largely been achieved by gene deletion through homologous recombination using a drug resistance marker. However, this approach is not feasible to study essential gene function. Here, we describe a method for the study of essential genes using a tetracycline-repressible promoter replacement system, which can be used to genetically repress essential genes in C. auris and, thus, study their function. This method provides a powerful approach to assess and characterize essential gene function in an emerging fungal pathogen.