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. 2021 Jun 3;11:11818. doi: 10.1038/s41598-021-91307-y

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© The Author(s) 2021

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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Application of electric field with µVS (eµVS) promotes intracellular delivery and TRAC-1 knockout. TRAC-1-targeting Cas9–RNPs were delivered to activated primary human CD3⁺ T cells via eµVS with an applied electrical field. Quantification of (A) percentages of TCR knockout cells and (B) viable T cells were quantified via flow cytometry at multiple timepoints post-transfection. TCR knockout levels were measured via TCRα/β staining and quantified as a percentage of TCRα/β-negative cells and the total number of live cells. eµVS with no applied electric field (0 kV cm⁻¹) and non-transfection samples served as controls. Data represent the mean ± SD of n ≥ 2 samples per condition and ≥ 2 independent experiments. Statistical analysis performed between groups indicated on graphs. Dot plots representative of replicate samples. P < 0.05 by unpaired, two-tailed, heteroscedastic t tests. Figure generated using Adobe Illustrator Creative Cloud (adobe.com/products/illustrator.html) and GraphPad Prism 9 (graphpad.com/scientific-software/prism/).