کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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6108686 | 1211191 | 2011 | 10 صفحه PDF | دانلود رایگان |

Background & AimsIn this study, adenoviral vectors encoding an antisense RNA complementary to the 5â² non-coding region (5â²NCR) of the HCV-genome were generated to inhibit HCV-RNA gene expression in cell culture and in vivo.MethodsFirst and second-generation (with E4-deletion) adenoviruses encoding the HCV5â²NCR in antisense direction (Ad-NCRas and Ad-E4del-NCRas) were generated. Inhibition of HCV gene expression was analyzed in hepatoma cells stably transfected with the HCV5â²NCR cDNA fused to the firefly luciferase gene (NCRluc), as well as in the HCV subgenomic replicon (genotypes 1b and 2a) and the fully infectious HCV JFH-1 cell culture systems. For in vivo experiments, an adenovirus encoding the NCRluc-gene was injected intravenously to achieve a NCR-dependent luciferase-expression in the liver of C3H/HeNcrl-mice.ResultsForty eight hours after transduction with GFP-encoding adenoviruses, >85% of HepG2-, CCL13-and Huh7-cells expressed GFP. Surprisingly, GFP-expression of E4-deleted adenoviruses was considerably reduced at the same MOI. Using antisense first-generation adenoviruses (Ad-NCRas), a significant inhibition of the 5â²NCR-dependent HCV-gene expression (54 ± 19% in HepG2-cells and 66.2 ± 15% in Huh7-cells) was achieved 48 h after transduction. In Huh7-cells containing the HCV subgenomic replicons and in infectious HCV JFH-1 cell cultures, adenovirus-mediated transcription of antisense 5â²NCR significantly blocked HCV-replication (40% and 76%, respectively). Corresponding to low transgene expression, the maximal inhibition reached with Ad-delE4-NCRas was 30%. In vivo, antisense adenoviral vectors also showed a significant inhibition (40%) of NCR-dependent luciferase expression compared to control adenoviruses (p <0.05).ConclusionsThe data indicate that HCV gene expression can be inhibited by antisense RNA encoding adenoviruses in the tested settings.
Journal: Journal of Hepatology - Volume 55, Issue 1, July 2011, Pages 19-28