eprintid: 10181415 rev_number: 9 eprint_status: archive userid: 699 dir: disk0/10/18/14/15 datestamp: 2023-11-20 14:57:27 lastmod: 2023-11-20 14:57:27 status_changed: 2023-11-20 14:57:27 type: article metadata_visibility: show sword_depositor: 699 creators_name: Malouf, Layla creators_name: Tanase, Diana A creators_name: Fabrini, Giacomo creators_name: Brady, Ryan A creators_name: Paez-Perez, Miguel creators_name: Leathers, Adrian creators_name: Booth, Michael J creators_name: Di Michele, Lorenzo title: Sculpting DNA-based synthetic cells through phase separation and phase-targeted activity ispublished: pub divisions: UCL divisions: B04 divisions: C06 divisions: F56 keywords: Synthetic cells, DNA nanotechnology, bottom-up synthetic biology, biomimetics, membrane-less organelles, phase separation, liquid-liquid phase separation, condensates, self-assembly note: © 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/). 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. date: 2023-11-09 date_type: published publisher: Elsevier BV official_url: https://doi.org/10.1016/j.chempr.2023.10.004 oa_status: green full_text_type: pub language: eng primo: open primo_central: open_green verified: verified_manual elements_id: 2107050 doi: 10.1016/j.chempr.2023.10.004 lyricists_name: Booth, Michael lyricists_id: MBOOT92 actors_name: Kalinowski, Damian actors_id: DKALI47 actors_role: owner full_text_status: public publication: Chem volume: 9 number: 11 pagerange: 3347-3364 issn: 2451-9294 citation: Malouf, Layla; Tanase, Diana A; Fabrini, Giacomo; Brady, Ryan A; Paez-Perez, Miguel; Leathers, Adrian; Booth, Michael J; Malouf, Layla; Tanase, Diana A; Fabrini, Giacomo; Brady, Ryan A; Paez-Perez, Miguel; Leathers, Adrian; Booth, Michael J; Di Michele, Lorenzo; - view fewer <#> (2023) Sculpting DNA-based synthetic cells through phase separation and phase-targeted activity. Chem , 9 (11) pp. 3347-3364. 10.1016/j.chempr.2023.10.004 <https://doi.org/10.1016/j.chempr.2023.10.004>. Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/10181415/7/Booth_1-s2.0-S2451929423005119-main.pdf