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. 2021 Dec 23;19(1):81–89. doi: 10.1038/s41592-021-01328-8

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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 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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Extended Data Fig. 4 — a, Evaluation of KO efficiency in polyclonal resting CD4⁺ T cells of single gRNAs for genes studied in functional assays. Cells were nucleofected with RNPs specific for the indicated genes, and the KO efficiency was evaluated by Illumina Miseq one week after nucleofection. Representative examples for each gRNA are shown. Together with TIDE analysis, this method was used to validate individual gRNAs. Depending on their efficiency, a combination of the two most potent gRNAs was used to obtain >95% KO efficiency, unless indicated otherwise. b, SAMHD1 decay in RNP-nucleofected cultivated for up to 6 weeks. Cells were nucleofected with either SAMHD1-gRNA2 + 3 or NTC and cultivated for the indicated period of time. Shown are immunoblots for either SAMHD1 or vinculin (loading control). One representative experiment is shown (n = 4).