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. 2023 Nov 13;26(12):2104–2121. doi: 10.1038/s41593-023-01480-8

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© The Author(s) 2023

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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Extended Data Fig. 1 — a, Schematic of generating human APOE4-R136S knock-in (E4-S/S-KI) mice using CRISPR/Cas-9-mediated gene editing. b, Schematic of gene editing strategy to generate the R136S mutation in human E4-KI mice (with the loxP sites, unused in current gene-editing strategy) and to generate APOE4-R136S (E4-S/S) hiPSC lines (without the loxP sites). c, DNA sequences of WT human APOE4 loci encoding for R136, designed sgRNA, and single-stranded oligodeoxynucleotides donor repair template encoding for S136 and silent mutation at PAM site for generating E4-S/S-KI mice. d, Sanger DNA sequencing of WT APOE4 and APOE4-S/S at and near the site encoding for residue 136 in E4-S/S-KI mice. e, Summary of on-target R136S editing in APOE4 and potential off-target mutation screening for knock-in mice. f, Representative immunofluorescent images of APOE (green) and GFAP (red) in CA1 hippocampal subfield in E4-KI and E4-S/S-KI mice at 12 months of age (scale bar, 50 μm). g, DNA sequences of WT APOE4 loci encoding for R136, designed sgRNA, and single-stranded oligodeoxynucleotides donor repair template encoding for S136 and silent mutation at PAM site for generating E4-S/S hiPSC lines. h, Sanger DNA sequencing of WT E4, E4-S/S, and E4-R/S at and near the site encoding for residue 136 in hiPSC lines. i, Summary of on-target R136S editing in APOE4 and potential off-target mutation screening in hiPSC lines. Experiments depicted in representative images in f were performed on n = 3 mice per genotype using 2 brain sections per mouse, with reproducible data. WT, wildtype; sgRNA, single guide RNA; ssODN, single-stranded oligodeoxynucleotide.