A study of ribozyme against hepatitis C virus in vitro

Objective To determine the nucleotide sites of hepatitis C virus (HCV), whi ch can be cleaved by ribozymes, and to obtain a highly effective, nontoxic and inexpensive antisense ribozyme specific for HCV. Methods Two effective ribozymes (HCRZNL and HCRZC), targeted to HCV 5' NC r egion and C region, were designed and synthesized. Eukaryotic expression ve ctors, pSV2-gpt, CD-SR alpha, containing either HCRZNC or HCRZC were constr ucted and transfected into MT-2 cells, which had been infected by HCV. Quan titative polymerase chain reaction (PCR) and hybridization were used to det ermine the effect of inhibition of HCV by ribozymes. Results HCRZNC and HCRZC suppressed the replication of HCV by 54.7% and 62. 1%, respectively. Furthermore, when both ribozymes were cotransfected into cells, they suppressed replication by 78.8%. Conclusion The specific antisense ribozymes, have strong inhibitory effects on the replication of HCV in cultured cells, and have better effect when u sed together.