RT info:eu-repo/semantics/article
T1 RNA interference (RNAi) for the silencing of extracellular serine proteases genes in Acanthamoeba: molecular analysis and effect on pathogenicity.
A1 Ortega Rivas, Antonio
A1 Lorenzo Morales, Jacob
A1 Foronda, Pilar
A1 Abreu Acosta, Néstor
A1 Ballart, David
A1 Martínez Carretero, Enrique
A1 Valladares Hernández, Basilio
A2 Didácticas Específicas
A2 Instituto de Enfermedades Tropicales y Salud Pública de Canarias
K1 Acanthamoeba
K1 RNAi
K1 Pathogenecity
K1 Serine protease
AB Silencing of extracellular serine protease genes was undertaken by interference RNA (RNAi). Chemically synthesized, small interfering RNA (siRNA) were highly specific and efficient in silencing the catalytic domain of extracellular serine proteases of Acanthamoeba. In order to confirmthe silencing phenomenon, the extracellular serine protease activities in RNAi-treated parasites were compared to non-treated parasites, usingzymographyprofiles,Acanthamoeba-conditionedmedium(ACM)proteaseactivity,cytotoxicityassaysandextracellularserine protease mRNA levels analysis. Zymography profiles showed a decrease in the extracellular protease levels in the moderate pathogenic and pathogenic strains, after treatment with siRNA. These results were supported after the ACM protease activity and CPE assays were performed in all studied isolates, showing a lower protease activity or cytotoxicity both in the pathogenic and moderate pathogenic strains treated with RNAi. These results support that extracellular serine proteases are directly involved in the pathogenesis and virulence of Acanthamoeba.
YR 2005
FD 2005
LK http://riull.ull.es/xmlui/handle/915/38858
UL http://riull.ull.es/xmlui/handle/915/38858
LA en
DS Repositorio institucional de la Universidad de La Laguna
RD 27-dic-2024