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. 2019 Feb 12;9:1847. doi: 10.1038/s41598-019-38526-6

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© The Author(s) 2019

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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Deep sequencing of CRISPR/Cas9 on-target sites. (a–c) Mutation rates and profiles of DSB repair outcomes in the target regions of HBV cccDNA isolated from HepG2-1.1merHBV cells. Each sgRNA (Sp1, Sp2, Sp3) has a mock control group, DMSO control group, and groups treated with NU7026 (see Supplementary Fig. S4 for results of treatment with other small molecules). Mock group was transfected with a Cas9 vector and non-targeting PCR product and treated with DMSO. Control groups were transfected with a Cas9 vector and an HBV-targeting sgRNA encoded by a PCR product, and treated with DMSO. PAM sequence for all sgRNAs was NGG (the strand complementary to the target DNA is provided). Regions containing the on-target sites were sequenced, and frequencies of insertions/deletions were calculated for sgRNAs Sp1 (d), Sp2 (e), and Sp3 (f) in cells treated with 3-aza, L755, B02, and Ad4E1B compared to mock control group and to DMSO control group. Number of indels per 1000 reads was counted for all experimental groups with corresponding sgRNAs.