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. 2021 May 6;49(9):4934–4943. doi: 10.1093/nar/gkab331

Figure 4.

Figure 4.

De novo MoiRNAiFold-generated toehold switch endogenous RNA sensors in vivo assays. (A) Schematic mechanism of toehold switches (ryhB sensor) regulating GFPmut3b-ASV translation upon endogenous ryhB small RNA (sRNA) as described in Green et al. (29). (B) In vivo assay in E.coli (strain K-12 substrain MG1655) grown in LB plus 100 μM FeSO4 was started by inducing GFP expression by 2,2-bipyridyl at different concentrations (0 mM -control- to 0.6 mM) during 1h and fluorescence in FL1 followed by flow cytometry. Data represent the AVG ± SEM of four experiments. (C) Bar graph representing the GFP fold-induction of ryhB sensor positive control and Moirai BD1–3 in the presence of 0.3 mM 2,2-bipyridyl. Data represent the AVG ± SEM of four experiments normalized to no 2,2-bipyridyl (0 mM) control. A paired t-test analysis shows significant differences between ryhB sensor control and ryhB Moirai BD1–3 at 0.3 mM 2,2-bipyridyl (**P < 0.01 and *P < 0.05, for Moirai BD1–2 and Moirai BD3, respectively). Raw data and complete statistical analysis and P values for each 2,2-bipyridyl concentration are shown in Supplementary Data 6.