Construction of RNAi lentiviral vector for MMP-2 gene

Objective To construct a lentiviral vector for MMP-2 gene RNA interference(RNAi).Methods The effective sequence of siRNA targeting MMP-2 gene was confirmed.Both sense and antisense Oligo DNA of the targeting sequence were designed,synthesized and cloned into the pLVTHM vector,which contained H1 promoter and green fluorescent protein(GFP).The resulting lentiviral vector containing MMP-2 siRNA was called LV2 siMMP-2,and it was confirmed by PCR and sequencing.293T cells were cotransfected with lentiviral vector LV2shMMP-2,pCMV2 dR8.74 and pMD2G and the lentivirus was produced.The titer of virus was tested according to the expression level of GFP.Results PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of MMP-2(LV2siMMP-2) producing MMP-2 siRNA was constructed successfully.The titer of concentrated virus was 8×1010TU/L.Conclusion The lentiviral RNAi vector for MMP-2 was constructed successfully,which provides possibility for further investigations of tumor biology.