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. 2017 Jun 8;7:3036. doi: 10.1038/s41598-017-02785-y

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© The Author(s) 2017

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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Off-target analysis. (A) The target sequence and ten of the predicted off-target sites for sgRNA91/Cas9. The PAM sequence is labeled in blue. (B) A total of ten potential off target loci (Lanes 1–10) were selected to examine off target effects in EGFP site-specific knock-in cell clone. In order to further confirm the specificity of sgRNA91/Cas9, ten EGFP site-specific knock-in cell clones (EK1-EK10) were selected randomly were evaluated by PCR and sequencing. WT: Wild-type. (C) T7E1 assays for the ten potential off-target sites. OT1-OT10 represents 10 potential off-target sites, and T represents target site.