Multi-signal embedded antigen receptor and expression gene thereof, NK (Natural Killer) cell modified by multi-signal embedded antigen receptor and application
A chimeric antigen receptor and gene expression technology, applied in genetically modified cells, cells modified by introducing foreign genetic material, animal cells, etc., can solve the problems of insensitive NK cell recognition process and many toxic and side effects of treatment.
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[0058] 1. Construction of lentiviral expression vector
[0059] 1.1c-Met scFv sequence
[0060] 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.
[0061] 1.2 Sequence synthesis and vector construction
[0062] According to the experimental requirements, three sequences were synthesized for the construction of lentiviral expression vectors:
[0063] Expression vector one (group one): pLenti7.3-γδTCR lentiviral expression vector, the sequence of which is shown in SEQ ID No.3, including signal peptide SP, TCRγ chain, and TCRδ chain.
[0064] Expression vector two (group two): pLenti7.3-c-Met-CAR lentiviral expression vector, the sequence is shown in SEQID No.4, including c-Met single-chain antibody (scFv), CD8 hinge region and transmembrane structure domain, 41BB intracellular domain and DAP12-ITAM domain.
[0065] Expression vector three (the third group)...
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