Fig. 5: Expression of RNA origami cytoskeletons in synthetic cells.
From: Genetic encoding and expression of RNA origami cytoskeletons in synthetic cells
a, Confocal time series of cytoskeleton-like iSpi RNA origami nanotubes (orange, λex = 488 nm) expressed inside a GUV (blue, membrane labelled with DiD, λex = 640 nm). Scale bars, 10 μm. b, RNA nanotube transcription inside of GUVs triggered by addition of rNTPs plotted over the first 3 h of expression (mean ± s.d., n = 6 GUVs). The data were extracted from confocal fluorescence time-lapse recordings (Supplementary Video 4). Background fluorescence outside of the GUVs is plotted as grey circles. c, Area fraction occupied by the RNA origami nanotubes plotted over time (n = 50, 6, 32 and 12 GUVs, left to right). d, Cortex formation with RNA origami nanotubes on the inner GUV membrane. Left: schematic representation of an RNA origami nanotube adhering to the biotinylated GUV membrane via a biotin aptamer attached to the iSpi tile38. Right: confocal 3D reconstruction of an RNA origami cortex on the inner GUV membrane after its expression. Scale bar, 10 μm. e, Distribution of RNA nanotubes with or without the biotin aptamer inside of the GUV. The distribution of the centre of mass of the iSpi fluorescence relative to the GUV centre is plotted (n = 78 (+) and 87 (−) GUVs; median, dashed lines; first and third quartile, dotted lines). f, GUV deformation caused by biotin aptamer-functionalized RNA origami nanotubes. Scale bar, 10 μm. g, Quantification of GUV deformation. The circularity of deformed GUVs (n = 19) and GUVs not expressing nanotubes (n = 12) are plotted (mean ± s.d.). A parametric, unpaired t-test with Welch’s correction was performed. Two-tailed P value is marked with *** (P = 0.0002).
