Objective: To observe whether or not the small-interfering RNA (siRNA) that conjugated with human immunodeficiency virus type 1 (HIV-1) TAT47–57 peptides can enter Huh-7 cells and efficiently silence hepatitis C virus (HCV) infection in cell culture. Methods: siRNA targeting the highly conserved stem loop IV of the HCV 5′ untranslated region (5′UTR) was conjugated to TAT47–57 peptides via the crosslinker sulfosuccinimidyl-4-(p-maleimidophenyl)-butyrate, and then the conjugates were added to the Huh-7 cell culture. Firefly luciferase activity and HCV RNA were asssessed using a luciferase assay reagent and real-time reverse transcript polymerase chain reaction, respectively. Results: The expression of firefly luciferase in HCV replicons and the concentration of HCV RNA were downregulated by siRNA-TAT47–57, and siRNA-TAT47–57 mediated RNA interfering activity which was directly correlated with increasing concentrations of the siRNA-TAT47–57 conjugate used. Conclusion: Cell-penetrating peptides such as HIV-1 TAT are an effective method for the delivery of siRNA targeted at 5′UTR of HCV in mammalian cells.

Keywords:
Delivery systems, Gene silencing, Hepatitis C virus, HIV-1, Small-interfering RNA, TAT peptide
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© 2009 S. Karger AG, Basel
2009
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