Construction and expression of bicistronic lentiviral vector containing TFR and VEGF genes in endothelial progenitor cells

AIM:To investigate whether bicistronic lentiviral vectors containing TFR and VEGF genes can mediate the expression of TFR and VEGF genes in endothelial progenitor cells (EPCs) of Chinese mini-swine. METHODS: TFR, IRES and VEGF genes were cloned into pLenti-GFP-Neo to generate the lentiviral vector pLenti-GFP-TIR. We sucessfully cultured EPCs of Chinese mini-swine where we identified the expression of cell surface CD31 and Flk-1 by flow cytometry (FCM). The four-plasmid lentiviral vector system (pRsv-REV, pMDlg-pRRE, pMD2G and pLenti-GFP-TIR) was cotransfected into human embryonic kidney 293T cells using the Lipofectin 2000 method. The packaged virus was harvested 72 h later and EPCs were infected by lentivirus carrying TFR and VEGF genes. RT-PCR was used to detect expression of TFR and VEGF genes in the viral-infected EPCs. RESULTS: EPCs were successfully infected by the recombinant lentivirus. TFR and VEGF genes were detected in the infected EPCs by RT-PCR. CONCLUSION: The bicistronic lentiviral vector containing TFR and VEGF genes can effectively deliver genes into EPCs and express target genes, providing the basis for future research on cardiac molecular imaging of cell transplantation.