Fig. 5: Enhanced ribozyme cleavage by compartmentalization.

a Schematic drawing of the RNA cleavage reaction. The fluorophore labeled hammerhead substrate (HHS) is cut into two smaller pieces by the hammerhead ribozyme (HHE) and emits green fluorescence. The substrate contains a donor that is quenched by FRET when HHS is not cleavage. b Image sequence shows RNA cleavage process inside the phase-separated compartments. c Control group of the RNA cleavage reaction inside an evaporating droplet while without HHE added. There is only fluorophore labeled substrate (HHS) introduced into the droplets. d Fluorescence intensity evolution of experimental group (both HHS and HHE are introduced into the evaporating ATPS droplet; Solid line), no ribozyme control group (only HHE is introduced into the evaporating ATPS droplet; Dashed line) as well as no ATPS control group (both HHS and HHE are introduced into the evaporating water droplet; Dash-dotted line). The intensity value is obtained by gel analysis of a specified rectangular area (as shown in (b) and (c)) in the fluorescence images. Images are obtained over analysis of three independent trials with relative humidity ranging from 55 to 65%. Insert: Dimensionless productivities \({m}_{p}\) of ribozyme cleavage in the domain of dextran-rich compartment (red solid line) and that of water droplet (gray solid line) as a function of dimensionless time \({t}^{{\prime} }\). Source data are provided as a Source Data file. The scale bar is 1 mm.