Extended Data Fig. 2: Generation of CAR T cells with a CRISPR-based knockout of EP2 and EP4. | Nature Biomedical Engineering

Extended Data Fig. 2: Generation of CAR T cells with a CRISPR-based knockout of EP2 and EP4.

From: Ablation of prostaglandin E2 signalling through dual receptor knockout in CAR T cells enhances therapeutic efficacy in solid tumours

Extended Data Fig. 2: Generation of CAR T cells with a CRISPR-based knockout of EP2 and EP4.The alternative text for this image may have been generated using AI.

(a) Representative transduction efficiencies for murine T cells with the different Ep2 and Ep4 knockouts with the anti-EpCAM-CAR measured using flow cytometry by its mCherry-tag. Genomic DNA samples from human anti-HER2 CAR T cells with their respective EP2 and EP4 knockouts were submitted to MiSeq Illumina Sequencing. Percentages of in- and out-of-frame indels are shown for n = 3 human donors at the PTGER2 locus (b) and the PTGER4 locus (c) for both gRNAs. (d) T cells with EP2 and/or EP4 knockout were treated with 1600 ng/ml (murine) or 2000 ng/ml PGE2 (human). CREB phosphorylation was measured by flow cytometry. Depicted is the gating strategy for flow cytometry analysis and a representative result for all knockout conditions.

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