Extended Data Fig. 4: Impact of CRISPR-mediated knockout of target genes on functions of human anti-LeY CAR T cells.
From: Rewiring endogenous genes in CAR T cells for tumour-restricted payload delivery
Each of the 27 target genes of interest was knocked out in human anti-LeY CAR T cells via CRISPR/Cas9 gene editing. a-f. Human CAR T cells with the indicated gene KO were cocultured with OVCAR-3 (a, c, e) or MCF-7 (b, d, f) tumor cells for 24 h before the concentrations of IFNγ (a-b), TNF (c-d) and IL-2 (e-f) in supernatants were assessed. g-h. Human CAR T cells with the indicated gene KO were cocultured with 51Cr-labelled OVCAR-3 (g) or MCF-7 (h) tumor cells for 16 h before quantifying 51Cr release. i-j. Human CAR T cells with the indicated gene KO were maintained in culture with IL-2 for over 28 days and the numbers of CD8+ (i) and CD4+ (j) T cells were determined. (a-j) Data represent logFC values normalized to Mock CAR T cells pooled from n = 3 donors. One-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
