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. 2021 Mar 25;4:396. doi: 10.1038/s42003-021-01942-4

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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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Fig. 4 — a, b hSOD1 sequence for Cas9 gRNA is on the top. DNA sequence, chromosomal position, number of nucleotide mismatch and bulge (Mi/Bu), and number of the identified mutants among the total number of the sequenced clones (Mut/clone) in 15 genome-wide potential off-target sites are shown. PAM sequences are shown in green. Nucleotide mismatches are shown in red. Nucleotide in blue indicates DNA bulge and (−) in blue indicates RNA bulge. Two G1H/Cas9 mice were analyzed (#8190, 585 days: a; #7449, 196 days: b). c Editing events identified in off-target #1. Arrowheads indicate the Cas9 cleavage site in off-target #1. Individual edited events are labeled on the top of sequencing chromatograms. Among 132 clones, three mutant clones were identified (delG, delAGinsGG, and delGTinsGAGTGGTCA) in mouse #8190, and among 156 clones, two mutant clones were identified (delG and delAAGG) in mouse #7449. The sites of mutations are indicated by arrows.