Fig. 2: Formation of temperature-responsive synthetic cells.
From: Genetically programmed synthetic cells for thermo-responsive protein synthesis and cargo release
a, GUVs, the synthetic cell chassis, were formed using emulsion phase transfer: a lipid-in-oil emulsion containing aqueous droplets is driven through a lipid–aqueous interface to form a bilayer. The PURExpress CFPE system was included in the emulsion phase for encapsulation. b, Representative fluorescent and merged fluorescent–phase contrast images of synthetic cells after in situ expression of constitutive dGFP (not under RNAT control) at 42 °C. c, The distribution of fluorescence intensity of individual vesicles across the constitutively expressing population, normalized for background signal (Methods). n = 114 vesicles; box plot bounds the 25–75% data range with a central median line; x denotes mean signal; whiskers denote 1.5 times the interquartile range. Phase transfer does not allow precise control of encapsulation, leading to variation in expression efficiency (and, therefore, fluorescent signal intensity) across the population. d, Representative fluorescent images of synthetic cells containing dGFP under control of RNA3-1 after incubation for 2 h at 30 °C (left) and 42 °C (right). Dotted circles denote the location of fluorescing cells in the 30 °C sample. Cells incubated at 42 °C have stronger levels of dGFP expression compared to 30 °C due to the RNAT being switched ‘on’ at this higher temperature, enabling translation. e, Mean normalized signal intensity showed that cells incubated at 42 °C had significantly higher (21.03-fold) dGFP expression compared to those incubated at 30 °C (unpaired two-tailed t-test P = 3.91 × 10−5). n = 3 biological replicates with over 100 vesicles measured per replicate; x denotes mean signal; open circles denote mean signal of individual replicates. Full sample data can be found in Supplementary Fig. 4 and the Source Data file. fluo., fluorescence.
