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. 2017 Oct 6;7:12739. doi: 10.1038/s41598-017-10418-7

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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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Schematic outline of gene-targeting and telomere truncation of human chromosome 21 in CHO-K1 cells using CRISPR/Cas9. (a) Human chromosome 21 was truncated in the region of AP000167 with CRISPR/Cas9, and the circular I-EGFP-I-Bsd Fcy; Fur vector inserted, which contained two homologous recombination arms, the EGFP gene and the blasticidin-resistance gene. Primer pair #1 and 2 were designed to detect the occurrence of homologous recombination. (b) Human chromosome 21 was truncated in the region of AP001657 with CRISPR/Cas9, and a linearised pBS-TEL/Dq HisDv2 Fcy; Fur vector inserted, which contained a single homologous recombination arm, 1 kb of artificial telomere sequence and a histidinol dihydrochloride-resistant gene (hisD). Primer pair #3 was designed to detect correct homologous recombination and primer pairs #4, 5 and 6 were designed to detect deletion of chromosome regions. Ideally, primer pair #3 should produce a PCR amplicon and primer pairs #4, 5, and 6 should not produce any PCR amplicons.