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. 2020 Sep 23;18:131. doi: 10.1186/s12915-020-00866-5

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© The Author(s) 2020

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 — ACBE-mediated base editing effects with different spacer lengths. a Schematic design for different spacer length of EGFP-Stop2. The red sequence indicated the PAM. b Flow cytometry results for testing different spacer lengths of EGFP-Stop2, n = 3. Untreated cell samples served as control. c, d The sequences represented the serial sgRNAs with different spacer lengths at P53-G8 (c) and P53-G9 (d) loci. The red sequence indicated the PAM. e, f Summary of ACBE-mediated base editing patterns and efficiencies of all C and A in P53-G8 (e) and P53-G9 (f) sites using sgRNAs with different spacer lengths. All values and error bars above indicate mean ± s.e.m. of three independent experiments. The controls were untreated cell samples. g, h The frequencies of simultaneous A > G and C > T mutation for P53-G8 (g) and P53-G9 (h) loci with different spacer length gRNA