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Construction of mcDNA and CD44-CAR T cells. (A) Schematic representation of anti-CD44 CAR structure. (B) Schematic diagram of CD44-CAR mcDNA generation. Anti-CD44 scFv was cloned into the parental plasmid to prepare pMC.CMV-CD44-CAR. L-arabinose was added to induce site-specific recombination. Bacterial backbone was digested for degradation and CD44-CAR mcDNA was generated. (C) Transfection efficacy demonstrated by fluorescence microscopy images within 48h at 400× magnification.
