title: [Construction of a lentiviral vector for RNA interference of glycerol kinase gene in human hepatocytes in vitro].
abstract: OBJECTIVE
  To construct a lentiviral vector for RNA interference (RNAi) of human glycerol kinase (GK) gene to stably down-regulate GK expression in human hepatocytes.
  
  
  METHODS
  The 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.
  
  
  RESULTS
  The 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.
  
  
  CONCLUSIONS
  The lentiviral vector for RNAi of human GK gene has been successfully constructed, which can significantly down-regulate GK expression in human hepatocytes.
authors.affiliation: Department of Pharmaceutical Engineering, Tianjin University, Tianjin, China. bronya@126.com
authors.forename: Yue
authors.initials: Y
authors.lastname: Liu
authors.forename: Yue
authors.initials: Y
authors.lastname: Hao
authors.forename: Yiqun
authors.initials: Y
authors.lastname: Zhan
authors.forename: Wangxiang
authors.initials: W
authors.lastname: Xu
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Yang
authors.forename: Changhui
authors.initials: C
authors.lastname: Ge
mesh_terms: D002460:Cell Line; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D005991:Glycerol Kinase / Q000235:genetics*; D022781:Hepatocytes / Q000201:enzymology; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D010957:Plasmids; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection
chemical_list: D034741:RNA, Small Interfering; D005991:Glycerol Kinase