Author/Authors :
Moazeni, Maryam tehran university of medical sciences tums - school of public health - division of molecular biology, department of medical mycology and parasitology, تهران, ايران , Khorramizadeh, Mohammad Reza tehran university of medical sciences tums - school of advanced technologies in medicine - department of medical biotechnology, تهران, ايران , Teimoori-Toolabi, Ladan pasteur institute of iran - biotechnology research center - department of molecular medicine, ايران , Noorbakhsh, Fatemeh islamic azad university, varamin-pishva branch - department of biology, ايران , Fallahi, Ali Akbar tehran university of medical sciences tums - school of public health - division of molecular biology, department of medical mycology and parasitology, تهران, ايران , Rezaie, Sassan tehran university of medical sciences tums - school of public health, School of Advanced Technologies in Medicine - division of molecular biology, department of medical mycology and parasitology, department of Medical Biotechnology, تهران, ايران
Abstract :
Background: The most important virulence factor which plays a central role in Candida albicans pathogenesis is the ability of this yeast to alternate between unicellular yeast and filamentous hyphal forms. Efgl protein is thought to be the main positive regulating transcription factor, which is responsible for regulating hyphal-specific gene expression under most conditions. ALS3 is one of the Efgl-associated genes encoding a multi-functional adhesive polypeptide, which mediates adherence to diverse host substrates. In this study, the EFG1 gene was knocked down by using synthetic siRNA in C. albicans and the regulation in ALS3 as one of the Efgl -dependent genes was investigated. Method: The 19-nucleotide siRNA was designed based on cDNA sequence of EFG1 gene in C. albicans. Transfection was performed using modified- plyethylen glycol/LiAc method. To quantify the level of EFG1 and the hyphal-specific ALS3 gene expression, the cognate EFG1 and ALS3 mRNA were measured in C. albicans by quantitative real-time RT-PCR. Results: Fluorescent microscopy pictures indicated that transfection was performed successfully. Also, according to relative expression software tool, expression of EFG1 gene was decreased significantly with 500 nM siRNA as well as 1 μM siRNA (P 0.05). However, more significant down-regulations were observed in the expression of ALS3 in both concentrations of 500 nM and 1 μM siRNA (P 0.05). Conclusion: In conclusion, we demonstrated the down-regulation of ALS3 gene as a consequent of applying EFGI-specific siRNA in C. albicans. This may lead us to design anti-fungal-specific agents in order to face with C. albicans-associated infections.
Keywords :
ALS3 , Candida albicans , EFG1 , RNAi
