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. 2022 Apr 20;7:108. doi: 10.1038/s41392-022-00936-w

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© The Author(s) 2022

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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Fig. 1 — Design and characterization of WT-PE. a A diagram showing the putative action mode of WT-PE. Different from PE2, WT-PE uses a nuclease preserved spCas9 and introduces DSB in the target site (left panel). The broken non-target strand is then recognized by PBS of the pegRNA and then extended by RT domain in the guidance of RT template. The RT template is designed to contain aimed edit (in red) and a homologous arm (HA, in green) that is complementary to the PAM proximal end of the target strand (right panel). b HTS analysis of the editing outcomes of WT-PE. Three types of outcomes were observed through the HTS: accurate edits, edits containing indels and pure indels. The prime edits represented accurate edits+edits containing indels. Plots showed mean ± s.d. of three independent biological replicates. c Sequence alignment showing top 10 sequences of WT-PE outcomes in HEK2 locus. Desired sequence containing +1 A to T conversion served as a reference sequence with spacer and PAM sequences underlined. The position of DSB was labeled with a red dash. d Schematic diagram of three types of outcomes.