RNAi as a Tool to Study Virulence in the Pathogenic Yeast Candida glabrata
Journal article, 2019

The yeast Candida glabrata is a major opportunistic pathogen causing mucosal and systemic infections in humans. Systemic infections caused by this yeast have high mortality rates and are difficult to treat due to this yeast's intrinsic and frequently adapting antifungal resistance. To understand and treat C. glabrata infections, it is essential to investigate the molecular basis of C. glabrata virulence and resistance. We established an RNA interference (RNAi) system in C. glabrata by expressing the Dicer and Argonaute genes from Saccharomyces castellii (a budding yeast with natural RNAi). Our experiments with reporter genes and putative virulence genes showed that the introduction of RNAi resulted in 30 and 70% gene-knockdown for the construct-types antisense and hairpin, respectively. The resulting C. glabrata RNAi strain was used for the screening of a gene library for new virulence-related genes. Phenotypic profiling with a high-resolution quantification of growth identified genes involved in the maintenance of cell integrity, antifungal drugs, and ROS resistance. The genes identified by this approach are promising targets for the treatment of C. glabrata infections.

RNA interference

RNAi

virulence factors

gene library

macrophages

antifungal drugs

Candida glabrata

pathogenic yeast

Author

Olena Ishchuk

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Lund University

Khadija Mohamed Ahmad

Lund University

Katarina Koruza

Lund University

Klara Bojanovic

Lund University

Marcel Sprenger

Hans-Knoll-Institute (HKI)

Lydia Kasper

Hans-Knoll-Institute (HKI)

Sascha Brunke

Hans-Knoll-Institute (HKI)

Bernhard Hube

Hans-Knoll-Institute (HKI)

Friedrich Schiller University Jena

Torbjorn Sall

Lund University

Thomas Hellmark

Lund University

Birgitta Gullstrand

Lund University

Christian Brion

University of Strasbourg

Kelle Freel

University of Strasbourg

Joseph Schacherer

University of Strasbourg

Birgitte Regenberg

University of Copenhagen

Wolfgang Knechtts

Lund University

Jure Piskur

Lund University

Frontiers in Microbiology

1664-302X (ISSN)

Vol. 10 1679

Subject Categories

Infectious Medicine

Microbiology

Microbiology in the medical area

DOI

10.3389/fmicb.2019.01679

PubMed

31396189

More information

Latest update

12/16/2020