Premium
cyp51A gene silencing using RNA interference in azole‐resistant A spergillus fumigatus
Author(s) -
Mousavi Bita,
Hedayati Mohammad T.,
TeimooriToolabi Ladan,
Guillot Jacques,
Alizadeh Ahad,
Badali Hamid
Publication year - 2015
Publication title -
mycoses
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.13
H-Index - 69
eISSN - 1439-0507
pISSN - 0933-7407
DOI - 10.1111/myc.12417
Subject(s) - gene silencing , microbiology and biotechnology , rna interference , azole , chemistry , gene , aspergillus fumigatus , rna , biology , biochemistry , antifungal
Summary An increasing number of reports have described the emergence of acquired resistance of Aspergillus fumigatus to azole compounds. The primary mechanism of resistance in clinical isolates is the mutation of the azole drug target enzyme, which is encoded by the cyp51A gene. The aim of this study was to evaluate the impact of silencing the cyp51A gene in azole‐resistant A. fumigatus isolates. A 21‐nucleotide small‐interfering RNA (si RNA ) was designed based on the cDNA sequence of the A. fumigatus cyp51A gene. After silencing the cyp51A gene in germinated conidia (15, 20, 25 and 50 nM), azole‐resistant A. fumigatus was cultured on broth media and gene expression was analysed by measuring the cyp51A mRNA level using RT ‐ PCR assay. Hyphae were successfully transfected by si RNA and expression of the cyp51A gene was significantly reduced by si RNA at the concentration of 50 nM ( P ≤ 0.05). In addition, at this si RNA concentration, the minimum inhibitory concentration of itraconazole for the treated cells was decreased, compared with that for untreated control cells, from 16 to 4 μg/ml.