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. 2020 Jan 15;10:409. doi: 10.1038/s41598-019-57295-w

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© The Author(s) 2020

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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Steps in SDSA and ODN-determined location of the indels. SDSA repair proceeds via distinct steps, starting with resection to yield 3′ overhangs on both sides of the Cas9 induced DSB. The 3′ overhangs pair with the ODN templates and are extended by DNA synthesis copying template sequences. Bridging of the DSB is completed when the newly synthesized strands withdraw from the donor and anneal back at the locus. This results in both perfect and erroneous repair, with indels primarily on the 5′ or 3′-end, depending on the polarity of the ODN. The FLAG insert is shown in light blue, and indels are shown as red diamonds.