Construction of a lentiviral vector for RNA interference of glycerol kinase gene in human hepatocytes in vitro.
- Author:
Yue LIU
1
;
Yue HAO
;
Yiqun ZHAN
;
Wangxiang XU
;
Yongsheng YANG
;
Changhui GE
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line; Gene Expression Regulation; Genetic Vectors; Glycerol Kinase; genetics; Hepatocytes; enzymology; Humans; Lentivirus; genetics; Plasmids; RNA Interference; RNA, Small Interfering; genetics; Transfection
- From: Journal of Southern Medical University 2012;32(5):614-617
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a lentiviral vector for RNA interference (RNAi) of human glycerol kinase (GK) gene to stably down-regulate GK expression in human hepatocytes.
METHODSThe sequence of siRNA for GK interference were cloned into the pSicoR vector. Following packaging in 293T cells, the lentivirus was titrated using fluorescence activated cell sorting. Human hepatocyte L02 cells was infected with the lentivirus and the expression of GK was analyzed using Western blotting.
RESULTSThe lentiviral particle pSicoR-GK was successfully packaged with a virus titer reaching 3×10(7) pfu/ml. The expression level of GK protein was down-regulated to 20% of the control level in L02 cells infected with the lentivirus.
CONCLUSIONThe lentiviral vector for RNAi of human GK gene has been successfully constructed, which can significantly down-regulate GK expression in human hepatocytes.
