@article{discovery10181415, pages = {3347--3364}, note = {{\copyright} 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/).}, journal = {Chem}, number = {11}, year = {2023}, title = {Sculpting DNA-based synthetic cells through phase separation and phase-targeted activity}, month = {November}, volume = {9}, publisher = {Elsevier BV}, issn = {2451-9294}, keywords = {Synthetic cells, DNA nanotechnology, bottom-up synthetic biology, biomimetics, membrane-less organelles, phase separation, liquid-liquid phase separation, condensates, self-assembly}, author = {Malouf, Layla and Tanase, Diana A and Fabrini, Giacomo and Brady, Ryan A and Paez-Perez, Miguel and Leathers, Adrian and Booth, Michael J and Di Michele, Lorenzo}, url = {https://doi.org/10.1016/j.chempr.2023.10.004}, abstract = {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.} }