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. 2023 Aug 30;621(7978):404–414. doi: 10.1038/s41586-023-06496-5

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Extended Data Fig. 12 — a. Triple epitope-edited cells are resistant to FLT3/CD123/KIT triple specific CAR-T cells. Triple FLT3/CD123/KIT reporter K562 were edited as previously described for all 3 targets and then FACS-sorted to isolate single-, double- and triple-edited cells, along with unedited K562. Purified populations were then co-cultured at different effector:target ratios with FLT3/CD123/KIT triple specific CAR-T cells obtained by co-transducing healthy-donor derived with LV vectors encoding the 4G8, CSL362 and 79D CAR constructs. Untransduced T cells were also included as a control. Co-cultures were incubated for 48h and then analysed by flow. N = 4 technical replicates. b. Interferon-γ and TNFα production by T cells assessed by flow cytometry (% of IFNγ+ and TNFα+ T cells). One-way ANOVA with multiple comparison. c. From left to right, target cell viability (AnnexinV-7AAD-), T cell activation (CD69+), T cell degranulation (surface CD107a+), T cell proliferation (CellTrace violet dilution), Target antigen expression (% of triple FLT3+/CD123+/KIT+ cells). Top row reports conditions cultured with FLT3/CD123/KIT CAR-T cells, bottom row conditions cultured with untransduced T cells.