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. 2023 Jan 13;24:6. doi: 10.1186/s13059-022-02836-2

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© The Author(s) 2022

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — Validation of Cas12a-ABE activity in stable wheat plants. a Schematic of stable wheat plant transformation and genotyping. b, c Percentage of T0 wheat plants with A-to-G base editing at independent positions of the target as measured by NGS. Two Cas12a-ABEs (v9h and v11h) are compared at TS60-A (b) and TS112-A (c) (v9h: n=155; v11h: n=182 at TS60-A and v9h: n=156 ; v11h: n=182 at TS112-A). Percentage of T0 wheat plants with heterozygous (editing rates between 25 and 70%) and homozygous mutations (higher than 70%) are displayed. d Percentage of T0 wheat plants carrying at least one mutation in at least one subgenome at TS60 or TS112 loci (n=154–157 for v9h and n=182–184 for v11h). Asterisks depict a significant difference in editing efficiency between v9h and v11h as measured by z-score test for two proportions (*: p<0.05; **: p<0.01; ***: p<0.001). e, f Allelic frequency of base edited T0 wheat plants at TS60-A (e) (v9h: n=314 alleles; v11h: n=368 alleles) or TS112-A (f) (v9h: n=316 alleles; v11h: n=368 alleles). Base-edited nucleotides are marked in green. g Distribution of T1 Cas12-ABE transgene-free plants with A-to-G edits in the three subgenome targets (A, B, D). 12 independent T1 lines were analyzed. WT, wild type; HZ, heterozygous; HM, homozygous