FIG. 5.

Sequence specificity of esiRNAs. (A) Effect of genotype 1a-, 1b-, and 2a-specific esiRNAs on the replication of an HCV genotype 1b replicon. About 2 × 10⁵ cells of Huh-7 clone 9B (containing the replicon I₃₈₉/NS3-3′/LucUbiNeo-ET) were transfected with 1.5 μg of esiRNAs, as indicated, or were left untreated. Two days later, cells were lysed and luciferase activities were determined, at least in duplicate. RLU, relative light units. (B) Effect of genotype 1a- and 1b-specific esiRNAs on the replication of a chimeric HCV replicon with genotype 1a coding sequences. About 2 × 10⁵ cells of the Huh-7 clone A-3/3 (containing the replicon I₃₈₉/NS3-3′/H77/DR) were transfected with 1 μg of esiRNAs, as indicated, or were left untreated. Half of the cells were harvested 2 days later. The other half was passaged (split ratio, 1:4), seeded into new cell culture dishes, and transfected a second time or left untreated. Two days later, these cells were also harvested, RNAs were prepared, and the replicon copy numbers per microgram of total RNA were determined by quantitative RT-PCR. Hatched columns, viral RNA concentration in control cells; black columns, viral RNA concentration after the transfection of HCV-specific esiRNAs. The figure shows the results from a single representative experiment.