Vector construction and silencing effect of VEGF gene targeted small interfering RNA

AIM:To construct VEGF gene-targeted small interfering RNA(siRNA) and its expression vector; and to observe the expression level of its expression vector VEGF gene in human osteosarcoma MG-63 cell transfected by the vector. METHODS:First to design VEGF gene-targeted hairpin siRNA,then to synthesize two complementary oligo nucleotide strand,after annealing,to insert the two-strand oligo nucleotide into pSuper neo vetor,which was then restrictive enzyme digested and sequenced. After that we transfected human osteosarcoma MG-63 cell line with the vector using lipofectamine method. Finally,detected the mRNA expression level of VEGF gene through RT-PCR. RESULTS:Restrictive enzyme digestion and sequencing confirmed the vector containing siRNA was what we wanted; RT-PCR showed the expression level of VEGF gene in transfected MG-63 was decreased. CONCLUSION:We had constructed the correct VEGF gene-targeted siRNA and its vector,and the vector could obviously reduce the VEGF gene expression after transfecting.