Objective: Chimeric antigen receptor (CAR)-engineered T cells (CAR-T) can destroy tumor cells expressing targeted tumor-associated antigen.However," On target, off tumor" effect is one of the most major safety concerns in the clinical application.Here, we designed a dual signal CAR T cells (dsCAR-T) which specifically recognize and eradicate tumor cells that expressing both MUC1 (Mucin1) and EGFR family protein and thus reduce the off-target effects of CAR-T cells.Methods: The expression cassettes of MUC1-specific CAR (MUC1-CD3 ζ), EGFR family-specific CCR (chimeric costimulatory receptor, Herin-CD28-4-1BB) and dsCAR(MUC1-CD3 ζ-F2A-Herin-CD28-4-1BB) gene were constructed and cloned into lentivirus transfer plasmids, respectively.