Design and screening of RNA interference lentiviral vector of human VEGF

Objective: Design and screening of clip siRNA lentiviral vector to clarify its functions.Methods: Three gene sequences and one non-sence control gene sequence were consistent, and then were connected to linear plenti6.3-MIR carriers in order to construct the miRNA lentiviral vector plasmid.The plasmid was transformed to DH5α cells,and the sequence were verified; 293T cells were infected by the lentiviral vector plasmid with liposome mediated,lentiviral vectors were packaged,and the titer was measured.The human liver cancer cells MHCC97L were infected by the lentiviral vectors,and the jamming effects to VEGF expression were valued by Realtime PCR.Results: Three VEGF RNAi lentiviral vector plasmids were constructed successfully. The lentiviral vectors were packed successfully,and the virus titers were 3.23×109、3.3×109、3.73× 109 TU/mL.Compared to the others,the miR-200 sequence had more significant efficiency and could silence 72% of VEGF gene.Conclusion: The lentiviral RNAi vector of human VEGF gene is constructed successfully,and expressed effectively in MHCC97L cells.The optimal efficient siRNA sequence was screened out,which will provide experimental basis for researching the role of VEGF gene in the effect of antitumor drugs.