iCas9 HA-T2A-antiCD19scFv-CD28-CD3z−BEAM
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The inducible caspase-9-CD19 CAR construct consists of FKBP12 gene with mutation F36 to V coding two 12 kDa FK506 binding proteins with HA tag at the end fused to truncated Caspase9 (135-416; lacking pro-domain), followed by T2A peptide and then CD19scFv-CD28- CD3zeta without signaling peptide and with BEAM. This construct can be used as a negative control for PM-CAR1006-1M (it expressed same length as icasp-CD19 without tags but is nonfunctional protein). In the presence of dimerization agent, AP20187 or AP1903, caspase-9 is activated inducing cell killing. Introduction of inducible caspase-9 can be used for increasing safety of CAR-T cells in the presence of dimerization agent
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-80°C; liquid nitrogen
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Positive control measured with flow cytometry
Figure 2: Transduction efficiency of icasp9-HA-CD19scFv-CD28-CD3zeta-GGGGSFlag CAR-T cells and CD19-CAR-T cells.
Addition of chemical inducer of dimerization measured with flow cytometry
Figure 3: Addition of chemical inducer of dimerization (CID) AP1903 100 nM for 24 hours did induce CAR-T cell killing.
For Research Use Only. Not for use in diagnostic procedures.