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
1664302x (eISSN)
Vol. 10 1679Subject Categories
Infectious Medicine
Microbiology
Microbiology in the medical area
DOI
10.3389/fmicb.2019.01679
PubMed
31396189