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. 2021 Nov 19;11:22627. doi: 10.1038/s41598-021-01978-w

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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An FLT3-based assay providing further evidence that locating two nicks close together facilitates TPN-based targeted knock-in. (a) Experimental setup of targeted knock-in of an FLT3-ITD. The length of a homologous region in the donor plasmid (pcDNA/FLT3-ITD) is noted in parenthesis. ITD, internal tandem duplication. (b) The efficiencies of targeted knock-in of the 24-bp ITD in the K562 (left) and NALM-6 (right) cell lines determined by IDAA assays. IDAA, indel detection by amplicon analysis. Data represent mean and SEM values from three independent experiments.