We are developing a novel technology platform for simplified genome editing in intractable fungal pathogens. Focusing on the pervasive fungal pathogen Candida albicans, we are developing and optimizing a sophisticated CRISPR-Cas9-based system to create large-scale genetic deletion libraries. Using this strategy, we are able to rapidly and efficiently generate homozygous deletion mutants in this otherwise genetically intractable diploid pathogen. We are now exploiting this platform to create genetic deletion libraries, to study the regulation of fungal pathogenesis. We are using this system to analyze cellular signaling pathways involved in complex virulence processes, including cellular morphogenesis, biofilm formation, and antifungal drug resistance.