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. 2026 Feb 20;16:7463. doi: 10.1038/s41598-026-40642-z

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© The Author(s) 2026

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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Fig. 6 — Enrichment and analysis of base-edited hiPSC populations via XMAS-TREE. (a) Diagram of the XMASTREE system, illustrating the expression of an mCherry cassette followed by a stop codon (TGA) and a GFP cassette. Conversion of the adenine in the stop codon to guanine activates GFP protein expression. (b) Diagram showing the tiling of two sgRNAs across the target genomic region. (c) Schematic of the XMASTREE workflow for identifying and enriching adenine base-edited cells. Flow cytometry was used to isolate populations that are double-positive for mCherry and GFP signal for downstream sequencing. (d) Percentage of adenine-to-guanine conversions along the protospacer. (e) Editing efficiency of the target adenine. The “specified mutation without bystanders” reports only clean desired-only (“perfect”) alleles within the analysed window. (f) Percentage of indels observed with ABE8e-SpRY or ABE9-SpRY, using sgRNA1 or sgRNA2 in sorted cells. Horizontal bars represent the mean from two biological replicates. Parts of the schematics in c were created with BioRender.com.