High-throughput functional profiling of the human fungal pathogen Candida albicans genome

Candida albicans is a major fungal pathogen of humans. Although its genome has been sequenced more than two decades ago, there are still over 4300 uncharacterized C. albicans genes. We previously generated an ORFeome as well as a collection of destination vectors to facilitate overexpression of C. albicans ORFs. Here, we report the construction of similar to 2500 overexpression mutants and their evaluation by in vitro spotting on rich medium and in a liquid pool experiment in rich medium, allowing the identification of genes whose overexpression has a fitness cost. The candidates were further validated at the individual strain level. This new resource allows large-scale screens in different growth conditions to be performed routinely. Altogether, based on the concept of identifying functionally related genes by cluster analysis, the availability of this overexpression mutant collection will facilitate the character-ization of gene functions in C. albicans. (c) 2023 The Authors. Published by Elsevier Masson SAS on behalf of Institut Pasteur. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Automated summary

Despite the complete sequencing of the genome of Candida albicans over two decades ago, there are still over 4300 uncharacterized genes. In this study, a collection of over 2500 overexpression mutants was constructed and evaluated to identify genes with fitness costs when overexpressed. This new resource enables routine large-scale screens under different growth conditions and facilitates the characterization of gene functions in C. albicans.