Project description:Purpose: Evaluate the effect of terbinafine in T. rubrum gene expression using a co-culture model in conjunction with RNA-seq Methods: T. rubrum and HaCat cells line were co-cultured in RPMI medium supplemented with 5% of fetal bovine serum and treated with 0.0162 µM of terbinafine.The co-culture was incubated for 24 h at 37ºC in a humidified atmosphere containing 5% CO2 , followed RNA by extraction of both organisms, libraries construction and sequence. Results: Our data demonstrated the modulation of several T. rubrum genes involved in the biosynthesis of ergosterol in addition to the target gene of terbinafine that is already known, genes encoding MFS- and ABC-type membrane transporters that are associated with the phenomenon of multidrug resistance, genes that have been discussed in the literature as possible novel targets for antifungal compounds and genes which do not have a known function in T. rubrum yet, but have been modulated in response to the drug. Conclusions: RNA-seq sequencing of the T. rubrum co-culture in the presence of terbinafine showed that several genes of the ergosterol biosynthesis cascade were repressed. We highlight the induction of transmembrane transporters, which may be associated with the efflux of this allylamine. In addition, other genes that could be involved in the pathogenicity of T. rubrum were also modulated in the presence of the drug. In addition, we observed the modulation of hypothetical genes with still unknown functions in T. rubrum but that possess orthologs in other dermatophytes.
Project description:Conidia are considered to be the primary cause of skin and nail infections by Trichophyton rubrum. A cDNA microarray containing 10112 ESTs was developed and used to estimate the relative gene expression levels and changes of gene expression during conidial germination in time-course experiments. Keywords: time course
Project description:A cDNA microarray was constructed from the expressed sequence tags (ESTs) of different developmental stages, and transcriptional profiles of the responses to 5-Flucytosine were determined. Keywords: Expression profiling by array The expression profiles of Trichophyton rubrum treated with 5-Flucytosine were compared to those of Trichophyton rubrum without drug. Each treatment has three replicates.