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. 2022 Mar 25;7:85. doi: 10.1038/s41392-022-00898-z

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© The Author(s) 2022

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — Disruption of CD5 and CD7 genes in T cells with CRISPR/Cas9 system. a Representative dot plots showing expression of CD5 and CD7 in CD5CD7KO T and mock T cells determined 3 days post electroporation with Cas9 protein, CD5 sgRNA-7, and CD7 sgRNA-85 using flow cytometry. b Proliferation and cell viability of CD5CD7 knockout T cells. Knockout of Adeno Associated Virus Integration Site 1(AAVS1), a safe harbor locus was used as a control. c The populations of CD4⁺/CD8⁺ T cells in CD5CD7KO T and mock T cells. The data represents mean ± SD for three donors. ns, no significance, two-way ANOVA. d The frequencies of effector and effector memory (TEFF/TEM; CCR7⁻ CD45RA⁻), terminally differentiated effector memory (TEMRA; CCR7⁻ CD45RA⁺), naïve (TNAIVE; CCR7⁺ CD45RA⁺), and central memory (TCM; CCR7⁺ CD45RA^-) in CD5CD7KO T and mock T cells were measured using flow cytometry on days 7–10. The data represents the average from three donors. e Heatmap of selected genes related to T-cell activation, exhaustion, cytokine production, differentiation, and function-related genes