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. 2009 May;15(5):898–910. doi: 10.1261/rna.1268209

FIGURE 7.

FIGURE 7.

Silencing of mutant HCV sequences by tsiRNAs. (A) Construction of target vectors. Wild-type or mutant E2 and NS3 siRNA-binding sequences were fused and cloned into XhoI and NotI sites downstream from the RLuc gene of the psiCHECK-2 vector. (E2N) Wild-type siE2 and siN-target sequence; (Nm) wild-type siE2 and mutant siN-target sequence; (E2m) mutant siE2 and wild-type siN-target sequence; (E2Nm) mutant siE2 and siN-target sequence. (B) Specific inhibition of HCV target sequences by pGD-siRNA vectors, pGD-siE2 and pGD-siN, relative to a control pGD-siC. (C–F) RLuc/FLuc activity in cells co-transfected with a series of HCV tsiRNA-expressing variants in the presence of psiCHECK-E2N, psiCHECK-Nm, psiCHECK-E2m, or psiCHECK-E2Nm 2 d post-transfection. Relative luciferase activity of cells treated with pGD-tsiC was set at 100%. The values represent the means (+SD) from three replicate transfections.