Related Experiment Video

Updated: May 13, 2026

Small-Scale Plasma Membrane Preparation for the Analysis of Candida albicans Cdr1-mGFPHis
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Small-Scale Plasma Membrane Preparation for the Analysis of Candida albicans Cdr1-mGFPHis

Published on: June 13, 2021

Pooled CRISPRi screening reveals fungal-specific drug target candidates

Lauren F Wensing1, Philippe C Després1,2,3, Desiree Francis1

  • 1Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada.

Nature Microbiology
|May 11, 2026
PubMed

Insights

This study introduces pooled CRISPR interference (CRISPRi) screening for large-scale functional genomics in Candida albicans. This method efficiently identifies essential genes and drug targets, aiding the development of new antifungal therapies.

Area of Science:

  • Mycology
  • Genomics
  • Antimicrobial Resistance

Background:

  • Drug-resistant fungal infections and intrinsically resistant pathogens present significant clinical challenges.
  • Developing effective antifungals is difficult due to the close evolutionary relationship between fungi and mammals, complicating efforts to avoid toxic off-target effects.
  • Traditional methods for functional genomics in Candida albicans are labor-intensive and low-throughput, limiting the pace of discovery.

Purpose of the Study:

  • To adapt pooled CRISPR interference (CRISPRi) screening for high-throughput functional genomic analysis in Candida albicans.
  • To identify essential and dosage-sensitive genes in C. albicans, particularly those conserved in fungi but lacking human homologues.
  • To investigate the impact of environmental conditions and genetic backgrounds on gene essentiality and drug resistance.

Main Methods:

  • Adaptation and implementation of pooled CRISPR interference (CRISPRi) screening in the fungal pathogen Candida albicans.
  • Assessment of repression sensitivity for 130 essential genes conserved across fungi.
  • Systematic screening across ten diverse environmental conditions and two drug-resistant clinical isolates.

Main Results:

  • Identification of highly dosage-sensitive genes across multiple essential pathways in C. albicans.
  • Discovery of environment-dependent gene sensitivity, highlighting the role of specific conditions in fungal gene function.
  • Demonstration that many identified fitness defects are conserved across different genetic backgrounds, including drug-resistant clinical isolates.

Conclusions:

  • Pooled CRISPRi screening is a powerful and rapid tool for large-scale functional genomics in Candida albicans.
  • This approach facilitates the identification of novel antifungal targets and the understanding of drug resistance mechanisms.
  • The findings provide a foundation for developing targeted antifungal therapies with reduced toxicity.

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