Project description:Candida auris reference strain B11221 was spread on YPD plate supplemented with 8 μg/ml tunicamycin. Randomly 25 adaptors (T2080-T2104) were chosen. These adaptors and the parent were sequenced.
Project description:Candidozyma auris, previously known as Candida auris, is a recently found pathogenic yeast that causes systemic infections, showing a high mortality rate. Moreover, this species is highly resistant to the commonly used antifungal drugs and some of the strains are multiresistant. This fungus is also able to cause outbreaks in hospital settings. All of this has cause an alarm in the health care system. Therefore, finding alternative treatments for C. auris is critical. In this sense, our research group developed a monoclonal antibody (Ca37) against Candida albicans alcohol dehydrogenase enzyme (Adh) that successfully reduced the growth of the fungus in vitro and also showed a protective effect in vivo. Due to the high homology between both fungal alcohol dehydrogenases, we wanted to test the effect of Ca37 over C. auris. In order to do this, first we assessed if the monoclonal antibody also recognized C. auris Adh by sequencing the spots detected by two dimensional western blot when using the monoclonal antibody as first antibody. Two spots were detected in this western blot. The sequencing showed a mixture of proteins, but in both cases, Adh was one of the identified proteins.
Project description:Creation of a new library entries for Candida auris using MALDI Biotyper. Candida auris has a high genetic variability in the world, the identification of Colombian isolates is difficult using the main Bruker library. A new in-house library was created using Colombian isolated and was validated using 300 isolated strains
Project description:Candida auris (Candidozyma auris) is a yeast pathogen that poses a public health threat because of its ability to develop antifungal resistance. Notably, most C. auris isolates are resistant to fluconazole. The efflux pump Cdr1 is a key contributor of azole resistance in C. auris. In C. albicans, Cdr1 is regulated by the transcription factor Tac1, which has two orthologs in C. auris (Tac1a and Tac1b). While the role of Tac1b has been described, little is known about Tac1a. In this study, we characterized the respective roles of Tac1a and Tac1b in azole resistance. To investigate their transcriptional regulation and binding targets, we performed RNA sequencing and ChEC-seq (Chromatin endogenous cleavage sequencing) for both transcription factors. RNA sequencing was carried out by comparing hyperactivated mutants (TAC1a-HA or TAC1b-HA) with the wild-type strain IV.1. ChEC-seq was performed in strains carrying endogenous C-terminal MNase fusions (TAC1a-MNase and TAC1b-MNase) and in a control strain expressing ectopically MNase (Free MNase). Both Tac1a and Tac1b mediate azole resistance in C. auris via regulation of Cdr1, with overlapping downstream targets and evidence of autoregulation.
Project description:Comparative transcriptomic profiling of Candida auris White and Brown cells from two clinical strains (1133/P/13R and 462/P/14) and their HYR3, WOR1, and MSN4 null mutants
Project description:Candida auris occupies similar niches in various infections as Pseudomonas aeruginosa; however, the details of their interspecies communication remain largely unknown. To gain deeper insights into this bacterial–fungal relationship, phenotypic and transcriptomic analyses were conducted in the presence of the primary P. aeruginosa quorum-sensing molecule, 3-oxo-C12-homoserine lactone (HSL), against C. auris, with the results compared to those of C. albicans. We demonstrated a significant HSL-induced reduction in adhesion of C. auris cells at 100- and 200-μM concentrations. Furthermore, HSL exposure reduced intracellular iron and zinc levels and modulated C. auris metabolism toward beta-oxidation, which may be associated with the observed reduction in in vivo virulence at lower HSL concentrations compared with C. albicans. RNA-sequencing transcriptome analysis revealed 67 and 306 upregulated genes, as well as 111 and 168 downregulated genes, in response to 100 and 200 μM HSL, respectively. We identified 45 overlapping upregulated and 25 overlapping downregulated genes between the two HSL concentrations. Our findings indicate that HSL-induced effects are not specific to C. albicans; additionally, several characteristics are present in C. auris but not in C. albicans following HSL exposure. Similar to other Candida-derived C12 compounds (e.g., farnesol), HSL reduces several C. auris survival strategies, which may significantly influence the nature of P. aeruginosa–C. auris co-habitation.
Project description:Candida auris clade III isolate B12039 was spread on YPD plate supplemented with 128 µg/ml fluconazole. Randomly 39 adaptors were chosen for further analysis. We did sequencing of them as as well as the parent.
