Inhibition of HBV core antigen gene expression in human embryonic kidney cell line AD293 by plasmid-based RNAi.
- Author:
Ning XU
1
;
Zheng-gang YANG
;
Hai-hong ZHU
;
Hang-ping YAO
;
Xiao-li HOU
;
Wei WU
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line; Embryo, Mammalian; Gene Expression Regulation, Viral; Green Fluorescent Proteins; biosynthesis; genetics; Hepatitis B Core Antigens; biosynthesis; genetics; Humans; Kidney; cytology; virology; Plasmids; Polymerase Chain Reaction; RNA Interference; physiology; RNA, Messenger; genetics; Transfection
- From: Journal of Zhejiang University. Medical sciences 2005;34(2):104-109
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo inhibit HBV core antigen gene expression with plasmid-based RNAi.
METHODSThe 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.
RESULTSHBV 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.
CONCLUSIONEffective 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.
