Establishment of a cell model supporting HCV 1b subgenomic replication

Objective To establish a cell model for supporting HCV 1b subgenomic replicon from Chinese population.Methods Plasmid containing HCV 1b subgenomic replicon was constructed. HCV subgenomic replicon RNA was obtained by transcription and then delivered into Huh-7.5.1 cell by electroporation. After screening the cells in the medium containing G418,we detected the HCV RNA in the obtained cell colonies by RT-PCR and the HCV NS5B protein by western blot. The cell colonies which contain HCV RNA were treated by IFN in order to observe the diminished level of HCV RNA.Results In a period of 4~6 weeks after transfection,visible colonies were stained by crystal violet. The HCV subgenomic replicon RNA NS4B was detected by RT-PCR and the HCV NS5B protein was detected by western blot. After treatment by IFN,HCV RNA diminished obviously. Conclusion We have successfully established a cell model for supporting HCV 1b subgenomic replicon of Chinese population replication in vitro. This cell model is a useful tool for the study on HCV pathogenesis,the screening of antiviral drugs and the development of vaccines.