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. 2017 Nov 8;7:15044. doi: 10.1038/s41598-017-15407-4

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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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Scarless genome editing in HEK239 cells modeling Gastric Cancer novel VUSs, a single-base substitution (PALB2-SNV) and a 9-base deletion (PALB2-DEL) in the PALB2 gene. (a) Timeline of CRISPR-Cas9 mediated HDR followed by KASP clonal genotyping pipeline to isogenic clones. (b,c) CRISPR-Cas9 HDR strategy and KASP mutation detection probes with mutation of interest (black nucleotides), guide RNA (red line), PAM site (red nucleotides), cleavage sites (red triangles) and 127-bp single-stranded asymmetric donor template for SNV and 9-base deletion respectively. (d,e) Isogenic clone plating in 96-well plate format and KASP genotyping cluster output of single-cell clones color-coded by genotype, Mut/Mut (blue), Mut/WT (orange), WT/WT (green) and no template controls (black) for SNV and 9-base deletion respectively.