Stably Silencing of CDB1 Expression by Small Interfering RNAs Targeting 31-NTR Inhibits HCV Infection

Background and aims: hepatitis c virus (hcv) entry, as the first key step during virus infection, includes cellattachment, interactions of virus envelope proteins with receptors or co-receptors on cell surface, membrane fusion andso on. human c081 is identified as an important receptor for hcv, which is known to interact with hcv e2 protein. theobjectives of this study were to further determine the association between c081 and hcv cell entry and provideinformation for the exploring of new anti-hcv agents.methods: in this study, the recombinant plasmid pegfp-c081 was firstly constructed using green fluorescence protein(gfp) as reporter gene. six small interfering rnas (sirna) targeting the open reading frame (orf) or 3'-nontranslatedregion (3'-ntr) of c081 genome were transcribed in vitro and transfected into pegfp-c081 expressing cells for thescreening of silence effect. the most effective sirna was selected to construct the short hairpin rna (shrna) expressingplasmid pgcsi-c081, which was stably transfected into huh7.s cells. after screening by g418, two cellclones with the c081 expression levels mostly reduced were infected with hcv pseudoparticles (hcvpp) or cell culturederived infectious hcv (hcvcc), while the cells stably transfected with an irrelevant sirna were used as negativecontrol.results: our results showed that the huh7.s cells where c081 was silenced were completely resistant to infection byhcvpp or hcvcc, while those cells stably transfected with an irrelevant sirna were sensitive to hcv infection.conclusions: these data underscore the importance of c081 as a receptor for hcv, and provide an approach forinvestigating the association between c081 and hcv cell entry, which may be of potential value in the development ofnovel prophylactic or therapeutic agents for hcv infection.