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. 2023 Jun 8;14:3369. doi: 10.1038/s41467-023-39137-6

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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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Fig. 4 — a Illustration of split circular TJ-petRNA. The prime editing template RNA (petRNA) sequence carrying an RTT-PBS sequence and an MS2 stem-loop aptamer, and circularized via a permuted group I catalytic intron. Yellow: circularization sequence. b Model of split circular petRNA function in PE. c Urea polyacrylamide gel showing split circular TJ-petRNA after splicing, RNase H, and RNase R digestion. Linear, but not circular, RNA is digested by RNase R. Experiments were done two times, and one is shown. d Editing efficiency of split circular TJ-petRNA at the AAVS1 locus. Synthesized sgRNAs and in vitro transcribed split circular petRNA were co-transfected with nCas9 and MCP-RT mRNA in HEK293T cells. For comparison purposes, HEK293T cells were transfected with PE2, TJ-pegRNA, and nicking sgRNA plasmids. FL-pegRNA: in vitro transcribed full-length TJ-pegRNA. Results were obtained from three independent experiments, shown as mean ± s.d.