FIGURE 3.

Fast prototyping de-novo designed sensors in low-cost optimized cell-free systems. (A) Scheme representation of fast prototyping de-novo designed RNA toeholds sensors against synthetic fragments of PVY virus. (B) PCR-purified transcriptional units (at 10 nM final concentration) encoding for PVY RNA toehold sensors were incubated with synthetic RNA direct or RNA reverse complementary (RC) (at 300 nM final concentration) and absorbance at 570 nm was measured in plate reader. Endpoint ON/OFF absorbance was calculated at 200 min with respect to an untriggered control. Heatmap values correspond to the average of six experimental replicates from two PCR amplifications performed on different days. (C) PVY Sensor 1 and ZIKV Sensor 27 encoded in plasmids (at 3.5 nM final concentration) were incubated with ZIKV Trigger 27 RNA or with PVY Trigger RNA (at 480 nM final concentration). Plotted values correspond to arithmetic mean and standard deviation of two independent experiments.