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

Fig. 6. Regulation of genome editing by CaVT-mediated translational activation and repression.

Fig. 6

a Schematic diagrams of RNA circuits to regulate gene knockout. EGFP knockout is induced by only the translational activation of Cas9 (top) or both the translational activation of Cas9 and translational repression of AcrIIA4 (bottom). b, c The percentage of EGFP-negative cells transfected with RNA circuits to regulate EGFP knockout. HeLa-EGFP cells were co-transfected with 1xMS2(U)site2-SpCas9 mRNA (cap analog: A-cap), 2xScMS2(C)-AcrIIA4 mRNA (cap analog: ARCA), CaVT mRNA, and EGFP-targeting sgRNA. For positive and negative controls, 1xMS2(U)site2-SpCas9 mRNA (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. The bar graph shows the average of three independent experiments (mean ± SD) (b). Representative histograms (c). ***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.