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Illustration of CAR construct
Figure 1: CAR-T cells expressing the above construct are available from Promab targeting CD19 with inducible
Positive control measured with flow cytometry
Figure 2: Flow cytometry with Fab Ab showing expression of scFv icasp9-CD19 CAR T-cells.
Negative control measured with flow cytometry
Figure 3: T cells that have not been transfected.
The inducible caspase-9-CD19 CAR construct consists of FKBP12 gene with mutation F36 to V coding two 12 kDa FK506 binding proteins fused to truncated Caspase9 (135-416; lacking pro-domain), followed by T2A peptide and then CD19scFv-CD28-CD3zeta. Administration of chemical inductor of dimerization (CID) CID (AP1903 or AP20187) results in dimerization of caspase 9 leading to an activated form of the molecule, which then initiates an apoptotic signaling leading to apoptosis of the transduced cell. Introduction of inducible caspase-9 is used for increasing safety of CAR-T cells by decreasing cytotoxicity of CAR-T cells in the presence of dimerization agent.