Chimeric antigen receptor with safety switch and its expression gene, its modified NK cell and application
A chimeric antigen receptor and gene expression technology, applied in the field of NK cells and applications, chimeric antigen receptors and their expression genes, can solve the problems of poor safety performance of NK cells, many side effects of treatment, and poor specificity
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[0057] 1. Construction of c-Met chimeric antigen receptor lentiviral vector with EGFRt switch
[0058] 1.1 c-Met scFv sequence
[0059] Using the anti-c-Met monoclonal antibody sequence screened by our laboratory, the VH and VL sequences were selected for the design of the scFv sequence in the CAR vector.
[0060] 1.2 Synthesis of c-Met CAR sequence with EGFRt switch
[0061] According to the sequence of each component of CAR, the full-length sequence contains signal peptide SP, c-Met single-chain antibody (scFv), CD8 hinge region and transmembrane domain, 41BB intracellular domain and DAP12-ITAM domain, T2A, Signal peptide SP, EGFRt, collectively referred to as SP-c-Met scFv-CD8-41BB-DAP12-T2A-SP-EGFRt, the structure diagram is as follows figure 1 shown. Restriction sites EcoR I and XhoI were added at the 5' and 3' ends of the entire framework, respectively, and Nanjing GenScript Biotechnology Co., Ltd. was entrusted to synthesize the entire expression cassette. The synthe...
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