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. 2022 Mar 14;13:1318. doi: 10.1038/s41467-022-28884-7

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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. 3 — a Heatmap of fraction active values at all NGNN PAMs. Nucleotides 1 and 4 are along the x-axis, nucleotides 2 and 3 along the y-axis. b Table of fraction active values for each PAM-flexible variant binned by activity bin. c Comparison of xCas9-3.7 (left) and Cas9-NG (right) to Legut et al. 2020. Points are colored by PAM. PAM-mapping z-scored LFC values on the y-axis refer to data in the present study. d Comparison of Cas9-NG and SpG fraction active values. Points are colored by PAM. Dashed lines at 0.3 indicate the cutoff for intermediate PAMs. e ROC-AUC values by 5′-type for each PAM-flexible variant. True positives are guides targeting essential genes, false positives are guides targeting nonessential genes. Only active PAMs are considered in this analysis. Source data are provided as a Source Data file.