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. 2020 Mar 10;11:1297. doi: 10.1038/s41467-020-15061-x

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© The Author(s) 2020

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. 7 — a, b Apoptosis-inducing circuit regulated by miRNA. HeLa cells were co-transfected with 1xMS2(U)site2-Bax mRNA (cap analog: A-cap), 2xScMS2(C)-BclxL mRNA (cap analog: ARCA), the indicated CaVT mRNA, and the indicated miRNA mimic or inhibitor. For the positive control, 1xMS2(U)site2-Bax mRNA (cap analog: ARCA) was transfected. All mRNAs contained N1mΨ. One day after the transfection, the cells were stained with Annexin V and SYTOX Red followed by flow cytometry. A schematic diagram of the RNA circuit (a). The bar graph shows the average of three independent experiments (mean ± SD) (b). *P < 0.05, **P < 0.01, ***P < 0.001 compared to the negative control by ANOVA with Dunnett’s multiple comparison test (two-sided). Exact P values are shown in Supplementary Table 1. Source data are provided as a Source Data file. c, d Cell-selective regulation of EGFP knockout by a miRNA-responsive genome editing circuit. The indicated EGFP-expressing stable cell lines were co-transfected with 1xMS2(U)site2-SpCas9 mRNA (cap analog: A-cap), 2xScMS2(C)-AcrIIA4 mRNA (cap analog: ARCA), the indicated CaVT mRNA, and EGFP-targeting sgRNA. For the positive and negative control, 1xMS2(U)site2-SpCas9 (cap analog: ARCA) with or without EGFP-targeting sgRNA was transfected, respectively. All mRNAs contained N1mΨ. Five days after the transfection, EGFP fluorescence was analyzed by a flow cytometer. A schematic diagram of the RNA circuit (c). The bar graph shows the average of three independent experiments (mean ± SD) (d). ***P < 0.001 compared to the non-treated samples by ANOVA with Dunnett’s multiple comparison test (two-sided). Exact P values are shown in Supplementary Table 1. Source data are provided as a Source Data file.