proteinscience影响因子

Establishment of a Screening System for Selection of siRNA Target Sites Directed against Hepatitis B Virus Surface GeneXiu-Min ZHOU1,3, Ju-Sheng LIN1, Yi SHI3, De-An TIAN2, Huan-Jun HUANG2, He-Jun ZHOU1, and You-Xin JIN3*1Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China; 2Department of Gastroenterology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China; 3State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, ChinaAbstract To study the inhibitory effects of plasmid-derived small interfering RNA (siRNA) and synthetic siRNA on the expression of the hepatitis B virus surface (HBs) gene, three plasmid-derived siRNAs and one synthetic siRNA that complement the coding region of the HBs gene were The HBs expression plasmid pHBs-EGFP was also HeLa cells were co-transfected with pHBs-EGFP and the above siRNA The HBs mRNA quantities were measured by reverse-transcription PCR, and the level of HBs-EGFP fusion protein was quantified by fluorescent The concentrations of the hepatitis B virus surface antigen (HBsAg) derived from the culture supernatant of transfected HepG15 cells were measured by an enzyme-linked immunosorbent assay (ELISA) The results showed that the three plasmid-derived siRNAs and the synthetic siRNA can effectively reduce the quantities of HBs mRNA and The plasmid-derived siRNA psiRNA1 was found to be the most effective inhibitor of HBs It can inhibit HBs-EGFP expression by 3% and suppress HBs mRNA by 6% To further substantiate the above observations, psiRNA1 was transfected into HepG15 cells (an HBV secreting cell line) The transfections resulted in almost complete blockage of HBsAg production, whereas control vector-transfected cells secreted high levels of HBsAg 7 days post- In conclusion, our data suggests that RNA interference (RNAi) is an efficient approach for reducing the level of HBs transcripts and proteins and for suppressing HBsAg Key words RNA interference (RNAi); small interference RNA (siRNA); hepatitis B virus surface gene (HBs gene)