CSB Seminar Series – Rebecca Shapiro – University of Guelph

CSB Departmental Seminar

Rebecca Shapiro

Department of Molecular and Cellular Biology, University of Guelph

TITLE: CRISPR-based functional genomic tools and applications to understanding antifungal resistance and fungal virulence

ABSTRACT: Opportunistic Candida pathogens are a leading cause of fungal infections, and new functional genomic tools enable our ability to better study the biology of these important pathogens. Here, we describe the development and optimization of CRISPR-based tools for functional genomic analysis in Candida albicans. We have developed two powerful technologies: CRISPR interference (CRISPRi) and CRISPR activation (CRISPRa), for applications in Candida species. We demonstrate the ability of these systems to robustly repress or induce gene expression in C. albicans, including in drug-resistant clinical isolates, and further demonstrate the application of these systems to characterize factors involved in resistance to antifungal drugs and fungal virulence. Since, unlike classic CRISPR systems, these platforms do not require a DNA repair construct, the simplicity of this system lends itself to high-throughput strain generation. Using a highly efficient, high-throughput cloning strategy, we are able to efficiently and rapidly generate large numbers of fungal mutant strains that over- or under-express any gene of interest, providing a powerful new tool for functional genomic analyses in fungal pathogens. We are applying these CRISPR-based functional genomic libraries to study antifungal drug resistance on a large scale, and understand mechanisms of fungal-host colonization and virulence.

HOST: Shelley Lumba

LOCATION: Cell and Systems Biology, 25 Harbord Street, Suite 432

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