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. 2022 Sep 8;41(1):96–107. doi: 10.1038/s41587-022-01410-2

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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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Extended Data Fig. 8 — (a) On-target indel formation of wild-type Nme2 nuclease, eNme2-C nuclease, and eNme2-C.NR nuclease compared to SpRY nuclease and SpRY-HF1 nuclease at each of the four protospacer-matched sites that were subsequently evaluated in GUIDE-Seq. Each bar represents the observed indel formation of one replicate in U2OS cells. (b-e) GUIDE-Seq identified off-targets and associated read counts for Nme2 variants (top) or SpRY variants (bottom) at Site 3 (b), Site 4 (c), Site 5 (d), and Site 6 (e). The on-target protospacer is marked by a black dot for each site. Off-target thresholds were set at 8 mismatches with and NNN PAM for SpRY variants or 11 mismatches with an NNNNNN PAM for Nme2 variants).