Targeting the 5 untranslated Region ofHepatitis C virus by Intracellularly Expressed Small Interfering RNA

Abstract：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．an d we also constructed the expression vector that Can express the short interfering RNAs (siRNA) against the HCV 5 ．UTR．The 5 ．UTR．eGFP／luciferase an d the siRNA-prod ucing plasmid were cotran sfected into Hela cells，an d the inhibition efect was detected by the intensity of the fluorescence an d luminescence．Th e vesults showed that the light from the cotransfected cells was obviously weaker than the negative control both in quality an d in quan tities，while density of the cotran sfected cells had no diference with the control plasmid as detected by nucleus staining．TlliS work demonstratedthat certain siRNA Can targetthe 5 UTR ofHCV whilenotoxic efecthadbeenobserved in the cells．TlliS work is the basis for future research in which RNAi activity is supposed to be utilized in the gene therapy with the the HCV infection．The siRNA is expressed intracellularly by vectors instead of chemical synthesis，an d a new method Can be used as a model to quickly an d safely screen effcctive siI AstargetingHCV