Targeting The 5'-Non-translated Region Of Hepatitis C Virus By Intracellularly Expressed Small Interfering RNA

In this study, we inserted the 5'-untranslated region (UTR) of Hepatitis C virus (HCV) genome into the upstream of the reporter genes of enhanced green fluorescent protein (eGFP) and luciferase, and we also constructed the expression vector that can express the short interfering RNAs (siRNAs) against the HCV 5'-UTR. Then the 5'-UTR-eGFP/luciferase and the siRNA-producing plasmid were cotransfected into Hela cells, and the inhibition effect was detected by the intensity of the fluorescence and luminescence. It showed that the light density from the cotransfected cells was obviously weaker than the negative control both in quality and in quantities, and the density of the cotransfected cells had no difference with control plasmid as detected by nucleus staining. This work demonstrated that certain siRNA can target the 5'-UTR of HCV, and no toxic effect had been observed in the cells. This work is the basis for future research in which RNAi activity is supposed to be utilized in the gene therapy with the HCV infection. The siRNA is expressed intracellularly by vectors instead of chemical synthesis, and a new method can be used as a model to quickly and safely screen effective siRNAs targeting HCV.