Construction of shRNA Lentiviral Vector Targeting Human GPC3 Gene and Its Expression in HepG2 Cells

Objective To construct shRNA lentiviral vectors targeting human GPC3 gene and observe the transfection and expression level in HepG2 cells.Methods The siRNA sequences specifically targeting the GPC3 gene were designed and cloned into lentiviral vector pGLV3-GFP using DNA recombinant technique.The 293T cells were transfected by lipofectin reagent for lentiviral particles packaged,and viral titer was determined.GPC3 mRNA and protein expression were examined by RT-PCR and Western blot.Results The recombinant lentiviral vectors were successfully constructed and the virus reached a titer of 1×108 TU/mL.The expression of GPC3 was significantly decreased after infection with the lentivirus.Conclusion shRNA expressing lentiviral recombinants targeting the GPC3 gene were successfully constructed,which provides a basis for the further functional study of GPC3.