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. 2021 Aug 19;4:988. doi: 10.1038/s42003-021-02503-5

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© The Author(s) 2021

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. 3 — a gRNAs and their target sequences in the human KRAS gene. b, c, e Results of CRISPR interference-RPA (CRISPRi-RPA) of gDNA extracted from 293T (b, e) or HCT116 cells (c, e). A schematic diagram of CRISPRi-RPA is shown in Fig. 1b and Supplementary Fig. 3a. Amplified KRAS is indicated by an arrowhead. M, molecular weight marker. d, f DNA sequencing analysis of CRISPRi-RPA products. CRISPRi-RPA products from c and e were purified and sequenced using a forward primer. g Mechanisms by which the discrimination of a single-nucleotide difference is achieved by CRISPRi-RPA. h A schematic diagram describing the evaluation of genome editing events by CRISPRi-RPA. i Results of CRISPRi-RPA. j Results of DNA sequencing analysis. RPA and CRISPRi-RPA products from i were purified and sequenced using a reverse primer. The CRISPR cleavage site is indicated by vertical dotted lines. The positions where intact CDKN2A (p16) nucleotides were not detected in the CRISPRi-RPA product are marked with asterisks.