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. 2023 Jun 30;6:681. doi: 10.1038/s42003-023-05052-1

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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. 2 — a Schematic overview for correcting KRAS mutations in HEK293T/17-KRAS library cells. Lentivirus particles containing 12 types of KRAS mutant-target sequences and barcode sequences were delivered in HEK293T/17 cells to generate HEK293T/17-KRAS library cells. After editing KRAS mutations using universal pegRNAs, the KRAS-corrected cells were collected and subjected to NGS analysis to measure the KRAS correction frequencies of each KRAS mutation. b Nine universal pegRNAs containing various lengths of PBS and RTT and PE2-SpG were delivered in HEK293T/17-KRAS library cells, and the KRAS correction frequency of each KRAS mutation is shown in the heatmap. c Universal pegRNAs containing 13-nt PBS and 16-nt RTT were engineered to have two types of RNA motif, tevopreQ1 and tmpknot, and subjected to KRAS correction in PE2-SpG or PE3b-SpG systems. The KRAS correction frequencies are shown in the heatmap and box-whisker plot, respectively. The experiments were conducted with biological triplicate (n = 3).