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. 2018 Oct 9;8:15028. doi: 10.1038/s41598-018-33408-9

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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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(a) ESC/Blastocyst Approach in Mouse Embryonic Stem Cells. The mouse Bcl2l11 locus, a gene targeting vector (pTLD39), and the modified locus are shown. A gene-targeting vector/donor molecular was constructed placing a 25-kbp segment of the human BCL2L11 gene between mouse homology arms, placing removable selectable marker cassettes at each end of the human segment, and placing loxP sites around a 2.9-kbp segment of human DNA deleted in 12% of the East Asian population (vector names, blue pTLD labels; guide binding areas, orange stars; genotyping oligonucleotide binding sites, oTLD-labelled arrows; proximal junction on mouse locus, mPJ; distal junction on mouse locus, mDJ; proximal mouse/human junction, PJ; distal mouse/human junction, DJ; Targeted Locus Amplification capture amplimers; circled numbers). See text for details. (b) CRISPR/Zygote Approach in Mouse Zygotes. The mouse Bcl2l11 locus, a gene targeting vector (pTLD67), and the modified locus are shown. Vector is as in “a.” above after removal of the Neo^R and Puro^R selection cassettes that are not necessary with the CRISPR/Zygote Approach (vector names, blue pTLD labels; genotyping oligonucleotide binding sites, oTLD-labelled arrows; proximal junction on mouse locus, mPJ; distal junction on mouse locus, mDJ; proximal mouse/human junction, PJ; distal mouse/human junction, DJ; junction over the deletion, ΔJ). See text for details.