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. Author manuscript; available in PMC: 2021 Dec 1.
Published in final edited form as: Cell Immunol. 2020 Sep 16;358:104216. doi: 10.1016/j.cellimm.2020.104216

Figure 1. Lentiviral transduction in 293T cells with F8CAR and F8CAR-mFoxp3 transgenes.

Figure 1.

(A) Scheme of the FVIIICAR constructs. The FVIII-CAR constructs were made by in-fusion cloning incorporating an FVIII-specific scFv followed by intracellular portions of signaling moieties of immune receptors into a lentiviral vector. The signaling moieties were composed of CD28, 4-1BB, CD3 signaling domains. For the F8CAR-mFoxp3 construct, murine Foxp3 cDNA was fused at the end of signaling moieties via F2A peptide. (B) Binding of FVIII to FVIII-CAR on lentivirus transduced cells. Lentivirus carrying F8CAR and F8CAR-mFoxp3 transgene were produced to test the transduction efficiency in HEK 293T cells. To detect FVIII-specific scFv, the transduced cells were incubated with human FVIII protein followed by anti-FVIII antibody. Control showed that the un-transduced cells did not bind to FVIII protein and anti-FVIII antibody.