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Objective To determine the nucleotide sites of hepatitis C virus (HCV), which 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 region and C region, were designed and synthesized Eukaryotic expression vectors, pSV2 gpt, CD SR α, containing either HCRZNC or HCRZC were constructed and transfected into MT 2 cells, The Institute of Infectious Diseases, Zhejiang Medical University, Hangzhou 310003, China (Chen Z, Liu KZ and Dou J) The Institute of Microbiology, Lubeck Medical University, Germany (Dennin RH and Reinhard U) which had been infected by HCV Quantitative polymerase chain reaction (PCR) and hybridization were used to determine 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 used together
Objective To determine the nucleotide sites of hepatitis C virus (HCV), which 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 region and C region, were designed and synthesized Eukaryotic expression vectors, pSV2 gpt, CD SR α, containing either HCRZNC or HCRZC were constructed and transfected into MT 2 cells, The Institute of Infectious Diseases, Zhejiang Medical University, Hangzhou 310003, China (Chen Z, Liu KZ and Dou J) The Institute of Microbiology, Lubeck Medical University, Germany (Dennin RH and Reinhard U) which had been infected by HCV Quantitative polymerase chain reaction (PCR) and hybridization were used to determine 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 cotransfecte d 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 used together