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      Artificial cell synthesis using biocatalytic polymerization-induced self-assembly

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          Abstract

          Artificial cells are biomimetic microstructures that mimic functions of natural cells, can be applied as building blocks for molecular systems engineering, and host synthetic biology pathways. Here we report enzymatically synthesized polymer-based artificial cells with the ability to express proteins. Artificial cells were synthesized using biocatalytic atom transfer radical polymerization-induced self-assembly, in which myoglobin synthesizes amphiphilic block co-polymers that self-assemble into structures such as micelles, worm-like micelles, polymersomes and giant unilamellar vesicles (GUVs). The GUVs encapsulate cargo during the polymerization, including enzymes, nanoparticles, microparticles, plasmids and cell lysate. The resulting artificial cells act as microreactors for enzymatic reactions and for osteoblast-inspired biomineralization. Moreover, they can express proteins such as a fluorescent protein and actin when fed with amino acids. Actin polymerizes in the vesicles and alters the artificial cells’ internal structure by creating internal compartments. Thus, biocatalytic atom transfer radical polymerization-induced self-assembly-derived GUVs can mimic bacteria as they are composed of a microscopic reaction compartment that contains genetic information for protein expression upon induction.

          Abstract

          Enzyme-initiated polymerization-induced self-assembly has been used to generate various biomimetic structures. Now, myoglobin’s activity is used for biocatalytic polymerization-induced self-assembly to generate vesicular artificial cells. As various cargoes can be encapsulated during polymerization, these artificial cells are capable of protein expression and can act as microreactors for distinct enzymatic reactions.

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          Fiji: an open-source platform for biological-image analysis.

          Fiji is a distribution of the popular open-source software ImageJ focused on biological-image analysis. Fiji uses modern software engineering practices to combine powerful software libraries with a broad range of scripting languages to enable rapid prototyping of image-processing algorithms. Fiji facilitates the transformation of new algorithms into ImageJ plugins that can be shared with end users through an integrated update system. We propose Fiji as a platform for productive collaboration between computer science and biology research communities.
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            Liposomes and polymersomes: a comparative review towards cell mimicking

            Minimal cells: we compare and contrast liposomes and polymersomes for a better a priori choice and design of vesicles and try to understand the advantages and shortcomings associated with using one or the other in many different aspects (properties, synthesis, self-assembly, applications). Cells are integral to all forms of life due to their compartmentalization by the plasma membrane. However, living organisms are immensely complex. Thus there is a need for simplified and controllable models of life for a deeper understanding of fundamental biological processes and man-made applications. This is where the bottom-up approach of synthetic biology comes from: a stepwise assembly of biomimetic functionalities ultimately into a protocell. A fundamental feature of such an endeavor is the generation and control of model membranes such as liposomes and polymersomes. We compare and contrast liposomes and polymersomes for a better a priori choice and design of vesicles and try to understand the advantages and shortcomings associated with using one or the other in many different aspects (properties, synthesis, self-assembly, applications) and which aspects have been studied and developed with each type and update the current development in the field.
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              Mechanistic insights for block copolymer morphologies: how do worms form vesicles?

              Amphiphilic diblock copolymers composed of two covalently linked, chemically distinct chains can be considered to be biological mimics of cell membrane-forming lipid molecules, but with typically more than an order of magnitude increase in molecular weight. These macromolecular amphiphiles are known to form a wide range of nanostructures (spheres, worms, vesicles, etc.) in solvents that are selective for one of the blocks. However, such self-assembly is usually limited to dilute copolymer solutions ( 99% monomer conversion) at relatively high solids in purely aqueous solution. Furthermore, careful monitoring of the in situ polymerization by transmission electron microscopy reveals various novel intermediate structures (including branched worms, partially coalesced worms, nascent bilayers, "octopi", "jellyfish", and finally pure vesicles) that provide important mechanistic insights regarding the evolution of the particle morphology during the sphere-to-worm and worm-to-vesicle transitions. This environmentally benign approach (which involves no toxic solvents, is conducted at relatively high solids, and requires no additional processing) is readily amenable to industrial scale-up, since it is based on commercially available starting materials.
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                Author and article information

                Contributors
                andrea.belluati@tu-darmstadt.de
                nico.bruns@tu-darmstadt.de
                Journal
                Nat Chem
                Nat Chem
                Nature Chemistry
                Nature Publishing Group UK (London )
                1755-4330
                1755-4349
                4 December 2023
                4 December 2023
                2024
                : 16
                : 4
                : 564-574
                Affiliations
                [1 ]Department of Pure and Applied Chemistry, University of Strathclyde, Thomas Graham Building, ( https://ror.org/00n3w3b69) Glasgow, UK
                [2 ]Department of Chemistry and Centre for Synthetic Biology, Technical University of Darmstadt, ( https://ror.org/05n911h24) Darmstadt, Germany
                [3 ]GRID grid.8534.a, ISNI 0000 0004 0478 1713, Adolphe Merkle Institute, , University of Fribourg, ; Fribourg, Switzerland
                [4 ]Biozentrum, University of Basel, ( https://ror.org/02s6k3f65) Basel, Switzerland
                Author information
                http://orcid.org/0000-0003-2305-7272
                http://orcid.org/0000-0002-0858-9441
                http://orcid.org/0009-0002-9065-8262
                http://orcid.org/0000-0002-1610-1768
                http://orcid.org/0000-0001-6199-9995
                Article
                1391
                10.1038/s41557-023-01391-y
                10997521
                38049652
                13ae772f-033d-428f-af1b-cee907f30c00
                © The Author(s) 2023

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 3 May 2023
                : 30 October 2023
                Funding
                Funded by: FundRef https://doi.org/10.13039/100010665, EC | EU Framework Programme for Research and Innovation H2020 | H2020 Priority Excellent Science | H2020 Marie Skłodowska-Curie Actions (H2020 Excellent Science - Marie Skłodowska-Curie Actions);
                Award ID: 101032493
                Award Recipient :
                Funded by: National Centre of Competence in Research (NCCR) Bio-Inspired Materials
                Funded by: UK Engineering and Physical Sciences Research Council
                Categories
                Article
                Custom metadata
                © Springer Nature Limited 2024

                Chemistry
                molecular self-assembly,polymer synthesis,synthetic biology,origin of life,nanoscale materials

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