%N 11
%T Sculpting DNA-based synthetic cells through phase separation and phase-targeted activity
%V 9
%A Layla Malouf
%A Diana A Tanase
%A Giacomo Fabrini
%A Ryan A Brady
%A Miguel Paez-Perez
%A Adrian Leathers
%A Michael J Booth
%A Lorenzo Di Michele
%D 2023
%P 3347-3364
%J Chem
%K Synthetic cells, DNA nanotechnology, bottom-up synthetic biology, biomimetics, membrane-less organelles, phase separation, liquid-liquid phase separation, condensates, self-assembly
%L discovery10181415
%I Elsevier BV
%O © 2023 The Author(s). Published by Elsevier Inc.
This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
%X Synthetic cells, like their biological counterparts, require internal compartments with distinct chemical and physical properties where different functionalities can be localized. Inspired by membrane-less compartmentalization in biological cells, here, we demonstrate how microphase separation can be used to engineer heterogeneous cell-like architectures with programmable morphology and compartment-targeted activity. The synthetic cells self-assemble from amphiphilic DNA nanostructures, producing core-shell condensates due to size-induced de-mixing. Lipid deposition and phase-selective etching are then used to generate a porous pseudo-membrane, a cytoplasm analog, and membrane-less organelles. The synthetic cells can sustain RNA synthesis via in vitro transcription, leading to cytoplasm and pseudo-membrane expansion caused by an accumulation of the transcript. Our approach exemplifies how architectural and functional complexity can emerge from a limited number of distinct building blocks, if molecular-scale programmability, emergent biophysical phenomena, and biochemical activity are coupled to mimic those observed in live cells.