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. 2021 Mar 23;12:639818. doi: 10.3389/fimmu.2021.639818

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Copyright © 2021 Muller, Nguyen, Ferreira, Ho, Raffin, Valencia, Congrave-Wilson, Roth, Eyquem, Van Gool, Marson, Perez, Wells, Bluestone and Tang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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CAR-CD28 heterodimers reduce CD28 expression. (A) Editing strategy and homology-directed repair-mediated integration into the TRAC locus of various CD19 CARs using an AAV-6 transduction protocol. (B) The expression of Myc and CD28 in a representative example was analyzed 6 days after editing and the removal of beads. (C) The CD28 MFI ratio was calculated by dividing CD28 MFI of Myc⁺ cells by Myc⁻ cells in the same culture. Pooled data from three to four independent experiments across five unrelated donors are shown. Each dot represents one independent editing condition. The two-way ANOVA was used for statistical analysis. ***p < 0.001.