FIG. 4.

Polymerase II-derived transcripts are functional templates for minus-strand RNA synthesis. (A) Schematic diagram of polymerase II-directed HCV minigenome reporter constructs. The HCV minigenome containing the antisense sequence of Renilla luciferase and the EMCV IRES flanked by the 5′ end (1 to 377) and 3′ partial NS5B coding region (9067 to 9371)-connected 3′ UTR was placed immediately next to the cytomegalovirus transcription start site or combined with a hammerhead ribozyme and was followed by a minimal poly(A) or HDV ribozyme upstream of the unmodified bovine growth hormone poly(A). (B) Huh-NNRZ, Huh-7, and HepG2 cells were transfected with the indicated reporter vectors. Relative Renilla luciferase activities in the lysates were determined as described for Fig. 1. The columns and bars represent the means and standard deviations of two separate triplicate transfections.