Establishment Of Transgenic Cell Lines For HCV 5'NCR, C By Microinjection And Inhibition Of HCV Gene Expression By Hammerhead Ribozymes

Hepatitis C virus (HCV) is the major cause of posttransfusionn hepatitis but there is no specific and effective therapy for it so far. Gene therapy may be a novel potential strategy against the virus. Human and champanzee can be exclusively infected by HCV. The lack of available animal model and efficient cell culture system permissive for HCV infection and replication has limited the systematic study of the evaluation of antiviral agents aimed at blocking HCV gene expression. In this context, the establishment of continuous transgenic cell lines then becomes a key step for the study of gene therapeutic strategies.Microinjection technique is superior to the other methods for cell transfection in two aspects. Firstly, the cell micromanipulation of one-by-one transfection facilitates the clonization of cell. Secondly, exotic gene fragments can be directly transmitted into the nuclei of target cells and escape from digestion of cellular nuclease and then be prone to integrating into DNA of the cells in the way of single copy. In our studies, microinjection method was used to establish continuous transgenic cell lines for HCV 5'NCR and C gene in order to investigate further gene therapeutic strategies against hepatitis C.