Self-replicating RNA molecule from hepatitis C virus

摘要

A Hepatitis C virus (HCV) self-replicating polynucleotide molecule is described. It comprises a 5'-non translated region (NTR) wherein guanine at position 1 is substituted for adenine. The molecule is further described to comprise a 5'-NTR consisting of ACCAGC (SEQ ID NO. 8); a HCV polyprotein region coding for a HCV polyprotein; and a 3'-NTR region. A host cell transfected with the self-replicating polynucleotide molecule is an eukaryotic cell line, in particular a hepatic cell line. If the cell is a human cell it is located ex vivo. A RNA replication assay comprising the following steps is disclosed. The host cell is incubated under conditions suitable for RNA replication; the total cellular RNA from the cells is isolated; and the RNA is analysed to measure the amount of HCV RNA replicated. A method for testing a compound for inhibiting HCV replication involves the steps as described above for RNA replication assay. A further step is to compare the levels of HCV RNA cells in the absence and presence of the inhibitor. Reduced RNA levels indicates the ability of the compound to inhibit replication.