Objective       To construct lentiviral vector LV-EF1α-CEA-2A-GFP-WPRE carrying chimeric antigen receptor (CAR) targeting carcinoembryonic antigen (CEA)

and determine its expression and cytotoxic effect on the CEA positive tumor cell lines when the vector are transfected into T cells.       Methods        The sequences coding anti-CEA antibody were obtained with standard molecular cloning techniques from the plasmid encoding antibody of CEA

and then verified by sequencing. The CEA segment was then inserted into lentiviral vector by enzyme dig

estion. Native Western blotting and flow cytometry were used to analyze its expression

and cytotoxic assay was employed to detect the cytotoxicity against the CEA positive tumor cell lines when the vector was infected into T cells.       Results        The sequencing result indicated that the CEA-CAR sequences were correct

and the enzyme digestion result exhibited lentiviral vector LV-EF1α-CEA-2A-GFP-WPRE was constructed correctly. When the vector was infected into T cells

the CEA-CAR

+

T cells were 62.8% and effectively killed the CEA positive tumor cells (P0.01).       Conclusion        Lentiviral vector LV-EF1α-CEA-2A-GFP-WPRE carrying CAR targeting CEA is constructed successfully. The CAR can be expressed through the vector and efficiently kill the CEA positive tumor cells in T cells.