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. 2018 Jan 12;8:593. doi: 10.1038/s41598-017-18911-9

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© The Author(s) 2018

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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Schematic representation of non-homologous end joining (NHEJ)-based knock-in using versatile NHEJ-based knock-in modules for genome editing (VIKING). Step one: select the donor plasmid, which harbors the VIKING–gRNA1 sequence (Vector A). Any vector with a pUC backbone could be used as the donor vector without any customization from publicly available resources. Step two: construct the vector for cleaving the target genome (Vector C). Step three: simultaneously transfect the three vectors. Step 4: select cell lines harboring successful knock-in.