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• W3159454241 abstract "Abstract Candida auris is an emerging healthcare-associated pathogen of global concern. Although this organism does not display the same morphological plasticity as the related fungal pathogen Candida albicans , recent reports have identified numerous C. auris isolates that grow in cellular aggregates or filaments. However, the genetic circuitry governing C. auris morphology remains largely uncharacterized. Here, we developed an Agrobacterium-mediated transformation system to generate mutants exhibiting aggregating or filamentous cell morphologies. Aggregating strains were associated with disruption of homologs of Saccharomyces cerevisiae chitinase and chitin synthase regulatory proteins, including components of the Regulation of ACE2 Morphogenesis (RAM) pathway, while disruption of a homolog of the S. cerevisiae ELM1 gene resulted in a novel filamentous strain of C. auris . To facilitate targeted genetic manipulation, we developed a transiently expressed Cas9 and sgRNA expression system for use in C. auris . Transformation using this system significantly increased the efficiency of homologous recombination and targeted integration of a reporter cassette in all four clades of C. auris . Using this system, we generated targeted deletion mutants to confirm the roles of RAM and Elm1 proteins in regulating C. auris morphogenesis. Overall, our findings provide novel insights into the genetic regulation of aggregating and filamentous morphogenesis in C. auris . Furthermore, the genetic manipulation tools described here will allow for inexpensive and efficient manipulation of the C. auris genome. Importance Candida auris is an emerging and often multi-drug resistant fungal pathogen responsible for outbreaks globally. Current difficulties in performing genetic manipulation in this organism remain a barrier to understanding C. auris biology. Homologous recombination approaches can result in less than 1% targeted integration of a reporter cassette, emphasizing the need for new genetic tools specific for manipulating C. auris . Here, we adapted Agrobacterium-mediated transformation and a transient Cas9 and sgRNA expression system for use in forward and reverse genetic manipulation of C. auris . We demonstrated the efficacy of each system by uncovering genes underlying cellular morphogenesis in C. auris . We identified a novel filamentous mutant of C. auris , demonstrating that this organism has maintained the capacity for filamentous growth. Our findings provide additional options for improving the genetic tractability of C. auris , which will allow for further characterization of this emerging pathogen." @default.
• W3159454241 created "2021-05-10" @default.
• W3159454241 creator A5042778763 @default.
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• W3159454241 date "2021-04-30" @default.
• W3159454241 modified "2023-10-10" @default.
• W3159454241 title "Forward and Reverse Genetic Dissection of Morphogenesis Identifies Filament-Competent<i>Candida auris</i>Strains" @default.
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• W3159454241 doi "https://doi.org/10.1101/2021.04.29.442079" @default.
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