Fig. 2

Characterization of SWTs with different design constraints. (A) Schematic of SWT with different toehold and stem sequence design constraints. (B) Leakage of SWTs (normalized fluorescence of OFF state) with different toehold region. The SWTs composed of different toehold region were designed with 5% GC decrement, and the differences of leakage values between them were compared. (C) Characterization of SWTs with different toehold GC contents. Fold change was measured with templates for SWTs at 10 nM and templates for trigger RNAs at 20 nM. (D, E) Fluorescence characterization was performed on SWTs with different stem regions. The leaky expression levels were measured with templates for SWTs at 5 nM. (F) Schematic of S1 and S13, combining the optimized toehold sequence with the redesigned stem sequence. (G, H) Fluorescence characterization was performed on S1 and S13. The data in G and H are the comparison of the leakage value (G) and ON/OFF ratio (H), respectively. All data shown are n = 3 independent biological replicates. For data in B, G, and H, Welch’s t-tests were performed on each construct, *P < 0.05, **P < 0.01, and ***P < 0.001. Error bars represent the standard deviation (s.d.) of three biological replicates