Analysis of the genes contribution to azole resistance in section with the CRISPR-Cas9 technique.

Cyp51 contribution to azole resistance has been broadly studied and characterized in , whereas it remains poorly investigated in other clinically relevant species of the genus, such as those of section In this work, we aimed to analyze the impact of genes ( and ) on the voriconazole (VRC) response and resistance of and We generated CRISPR-Cas9 and knock-out mutants from strains with different genetic backgrounds and diverse patterns of azole susceptibility. Single gene deletions of genes resulted in 2 to 16-fold decrease of the VRC Minimum Inhibitory Concentration (MIC) values, which were below the VRC Epidemiological Cutoff Value (ECV) established by the Clinical and Laboratory Standards Institute (CLSI) irrespective of their parental strains susceptibilities. Gene expression studies in the tested species confirmed that participates more actively than in the transcriptional response of azole stress. However, ergosterol quantification revealed that both enzymes comparably impact the total ergosterol content within the cell, as basal and VRC-induced changes to ergosterol content was similar in all cases. These data contribute to our understanding on azole resistance, especially in non- species.