Establish a screening system for selection of mRNA target sites for HBsAg to construct siRNA with shRNA.
- Author:
Zheng-Gang YANG
1
;
Zhi CHEN
;
Ning XU
;
Qin NI
;
Xiu-Cheng PAN
;
Han-Ying JIN
;
Min-Wei LI
Author Information
- Publication Type:Journal Article
- MeSH: Gene Expression Regulation, Viral; Gene Silencing; Gene Targeting; methods; Hepatitis B Surface Antigens; genetics; metabolism; Humans; RNA Interference; RNA, Small Interfering; genetics; RNA-Induced Silencing Complex; genetics
- From: Chinese Journal of Hepatology 2004;12(9):515-518
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo find some effective short interfering RNA's sites targeting HBV surface gene sequence using shRNA expression vectors.
METHODSFour shRNA expression vectors targeting HBV surface gene sequence were constructed based on pAVU6 + 27 vector, and cotransfected into AD293 cells with HBs-EGFP fusion gene plasmid. The changes of HBs-EGFP image were detected by FACS and microscopy. The HBs-EGFP mRNA expression was evaluated by RT-PCR.
RESULTSFour shRNA expression vectors and HBs-EGFP fusion gene plasmid were successfully constructed. pAVU6 + 4sh579 vector inhibited the HBs-EGFP expression by 69.8% in AD293 and suppressed the HBs-EGFP mRNA expression by 74.6%.
CONCLUSIONSThe results showed that the 579 site of HBV surface gene sequence was an effective target and pAVU6 + 4sh579 vector could suppress the HBs-EGFP expression in AD293 cells
