Construction of Inactivation Mutant for Candida albicans CaHSL1 with the Novel CRISPR/Cas9 Approach and Its Phenotype Analysis

Xu Huihui1, Jiang Linghuo1,2*, Malcolm Whiteway3

1School of Biotechnology,/the National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China; 2School of Agricultural Engineering and Food Science, Shandong University of Technology, Zibo 255000, China; 3Department of Biology, Concordia University, Montreal, Quebec, Canada H4B1R6

Abstract: Calcium-homeostasis and calcium signaling pathway are important for the morphogenesis， drug tolerance and virulence of Candida albicans. In both C. albicans and Saccharomycese cerevisiae， the plasma membrane protein Rch1p is enriched at the bud neck between the mother cell and the daughter cell， while Hsl1p related to the control of cell cycle is also localized to the bud neck. In this study， we show that conditional repression of CaHSL1 expression or inactivation of CaHSL1 expression through the novel approach CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 leads to calcium sensitivity of cells. This calcium sensitivity was suppressed by the block of calcium signaling through cyclosporin A， the specific inhibitor of calcineurin. This suggests that calcium homeostasis and calcium signaling might be related to cell cycle control. In addition， the CRISPR/Cas9 mutant for CaHSL1 is sensitive to sodium dodecyl sulfate， fluconazole， ketoconazole， congo red， hygromycin B， caspofungin and anidualafungin， indicating that CaHsl1p is involved in the response of C.albicans cells to cell membrane and cell wall stresses.

CSTR: 32200.14.cjcb.2018.04.0010