Construction and Identification of Lentiviral Vector-mediated RNA Interference of VEGF Gene

Objective To construct a lentiviral vector-mediated RNA interference(RNAi) of VEGF gene and provide the basis for further experiments in vivo and in vitro.Methods A VEGF gene targeting short hairpin RNA(shRNA) was cloned into lentiviral expression vector pRNAT-U6.2/Lenti.The recombined vector pRNAT-shVEGF confirmed by PCR and sequencing,was cotransfected with the pPACKH1 packaging plasmids mixture into 293T cells by Lipofectamine 2000.Virus in the supernatant was collected and the virus titer was measured.Results PCR and DNA sequencing demonstrated that the inserted sequences were correct.The titer of virus was 3×107 pfu/ml.Conclusion The lentivirus RNAi vector targeting VEGF has been successfully constructed,which will provide a tool for the further study on function of VEGF gene in leukemia cells and gene therapy.