Fig. 2: Expression of RNA origami inside GUVs.
From: Genetic encoding and expression of RNA origami cytoskeletons in synthetic cells
a,b, Schematic illustration showing the external triggers for RNA origami expression by Mg2+ import using ionophores (a) or rNTP feeding via α-haemolysin pores (b). c, AFM micrograph of the pentagon-shaped RNA origami (3H-4DT-iSpi), composed of five subunits with three helices each. Scale bar, 50 nm. d, AFM micrographs of Mg2+-triggered 3H-4DT-iSpi RNA origami. RNA was incubated for 2 h at 1 mM Mg(OAc)2 (left) before transcription was triggered by increasing Mg(OAc)2 to 6 mM and incubated for another 2 h (right). Scale bars, 200 nm. Panels c and d are images from lock-in phase channel. The corresponding height images are shown in Supplementary Fig. 1. e, Fluorescence plate reader experiments tracking 3H-4DT-iSpi RNA origami production by measuring DFHBI-1T emission over time (λex = 488 nm, mean ± s.d., n = 3 wells). The orange arrow indicates the time point where the Mg(OAc)2 concentration was increased from 1 mM to 6 mM. f, Confocal overlay images of 3H-4DT-iSpi RNA origami (orange, iSpinach binds DFHBI-1T, λex = 488 nm) transcribed inside of a GUV (blue, membrane labelled with DiD, λex = 640 nm). Quantification in Supplementary Fig. 5. Scale bars, 20 μm. g,h, RNA origami transcription inside of GUVs triggered by addition of Mg2+ (g) or rNTPs (h) over time (mean ± s.d., n = 6 GUVs). The data were extracted from confocal fluorescence time-lapse recordings (Supplementary Videos 1 and 2). Empty grey circles denote the background fluorescent signal outside of the GUVs.
