Extended Data Fig. 12: One-step manufacturing approach for CRISPR-engineered armored CAR T cells.
From: Rewiring endogenous genes in CAR T cells for tumour-restricted payload delivery
a-b. Anti-LeY CAR T cells manufactured via retroviral transduction or CRISPR-mediated knock-in to the TRAC locus were engineered to express Gfp from either the NR4A2 or RGS16 locus. a. Flow cytometry plots from concatenated technical triplicates (left) and quantification of CAR expression represented as mean ± SD of technical triplicates (right) in CD8+ human CAR T cells. b. Quantification of GFP expression in CD8+ human CAR T cells upon in vitro stimulation with anti-LeY idiotype antibody, MCF-7 or OVCAR-3 tumor cells for 24 h, represented as mean ± SD of technical triplicates. c-d. Anti-LeY CAR T cells manufactured via CRISPR-mediated knock-in to the TRAC locus were engineered to express IL-12 or IL-2 from the NR4A2 or RGS16 locus, respectively, and assessed upon in vitro stimulation as per (b). c. Concentration of IL-12 or d. IL-2 produced by NR4A2/IL-12 or RGS16/IL-2 CAR T cells, respectively, represented as mean ± SD of technical triplicates. (c-d) Two-way ANOVA. **p < 0.01, ***p < 0.001, ****p < 0.0001.
