Construction of a lentiviral vector for RNA interference of glycerol kinase gene in human hepatocytes in vitro / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To construct a lentiviral vector for RNA interference (RNAi) of human glycerol kinase (GK) gene to stably down-regulate GK expression in human hepatocytes.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>The lentiviral vector for RNAi of human GK gene has been successfully constructed, which can significantly down-regulate GK expression in human hepatocytes.</p>