OBJECTIVE: To determine the effect of internal ribosome entry site (IRES)specific inhibitor RNA (IRNA) on viral gene expression mediated by HCV 5' noncoding region (NCR) in human hepatic carcinoma cell line. METHODS: Cotransfection of pCMVNCRluc containing 5'NCR-luc fusion genes and eukaryotic vectors of IRNA into human hepatic carcinoma cells (HHCC) was performed and the luciferase activity of reporter gene was measured by luminometer. RESULTS: pCMVNCRluc was efficiently expressed in HHCC. IRES specific IRNA significantly inhibited the expression of luc mediated by HCV IRES by 50% to 90%, but the mutant IRNA lost the activity of inhibition. CONCLUSION: IRNA can inhibit HCV IRES mediated gene expression in vivo.
OBJECTIVE: To determine the effect of internal ribosome entry site (IRES)specific inhibitor RNA (IRNA) on viral gene expression mediated by HCV 5' noncoding region (NCR) in humanhepatic carcinoma cell line. METHODS: Cotransfection of pCMVNCRluc containing 5'NCR-luc fusion genes and eukaryotic vectors of IRNA into humanhepatic carcinoma cells (HHCC) was performed and the luciferase activity of reporter gene was measured by luminometer. RESULTS: pCMVNCRluc was efficiently expressed in HHCC. IRES specific IRNA significantly inhibited the expression of luc mediated by HCV IRES by 50% to 90%, but the mutant IRNA lost the activity of inhibition. CONCLUSION: IRNA can inhibit HCV IRES mediated gene expression in vivo.