[Inhibition of HBV core antigen gene expression in human embryonic kidney cell line AD293 by plasmid-based RNAi].
Ning Xu, Zhenggang Yang, Haihong Zhu, Hangping Yao, Xiaoli S. Hou, Wei Wu
Abstract
Ning Xu, Zhenggang Yang, Haihong Zhu, Hangping Yao, Xiaoli S. Hou, Wei Wu
Abstract
OBJECTIVE: To inhibit HBV core antigen gene expression with plasmid-based RNAi. METHODS: The shRNA expression vector targeting HBV core antigen gene was designed and constructed. Human embryonic kidney cell line AD293 was co-transfected with HBcAg-EGFP fusion protein expression vector and shRNA expression vector transiently, and the cells without shRNA-transfection and with non-specific shRNA transfection were used as controls. Inhibitory effect of RNAi was detected by fluorescence-activated cell sorting (FACS) and real-time fluorescence quantificational RT-PCR. RESULTS: HBV core antigen gene expression in AD293 was inhibited by shRNA, with the maximal inhibition rate of 76 % measured by FACS and of 63.1 % by real-time PCR. CONCLUSION: Effective inhibition of HBV core antigen gene expression by plasmid-based RNAi provides an alternative for anti-HBV study in vitro, which has potential clinical application.
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OBJECTIVE: To inhibit HBV core antigen gene expression with plasmid-based RNAi. METHODS: The shRNA expression vector targeting HBV core antigen gene was designed and constructed. Human embryonic kidney cell line AD293 was co-transfected with HBcAg-EGFP fusion protein expression vector and shRNA expression vector transiently, and the cells without shRNA-transfection and with non-specific shRNA transfection were used as controls. Inhibitory effect of RNAi was detected by fluorescence-activated cell sorting (FACS) and real-time fluorescence quantificational RT-PCR. RESULTS: HBV core antigen gene expression in AD293 was inhibited by shRNA, with the maximal inhibition rate of 76 % measured by FACS and of 63.1 % by real-time PCR. CONCLUSION: Effective inhibition of HBV core antigen gene expression by plasmid-based RNAi provides an alternative for anti-HBV study in vitro, which has potential clinical application.
Key concepts: Small hairpin RNA, RNA interference, HBcAg, Transfection, Molecular biology, Biology, Antigen, Plasmid