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WIREs Nanomed Nanobiotechnol
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Cell mimicry as a bottom‐up strategy for hierarchical engineering of nature‐inspired entities

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Abstract Artificial biology is an emerging concept that aims to design and engineer the structure and function of natural cells, organelles, or biomolecules with a combination of biological and abiotic building blocks. Cell mimicry focuses on concepts that have the potential to be integrated with mammalian cells and tissue. In this feature article, we will emphasize the advancements in the past 3–4 years (2017‐present) that are dedicated to artificial enzymes, artificial organelles, and artificial mammalian cells. Each aspect will be briefly introduced, followed by highlighting efforts that considered key properties of the different mimics. Finally, the current challenges and opportunities will be outlined. This article is categorized under: Nanotechnology Approaches to Biology > Nanoscale Systems in Biology
Cartoon of a mammalian cell containing artificial enzymes (AEs), artificial organelles (AOs) as well as natural organelles. Artificial biology aims to mimic functional and structural aspects of natural cells ranging from enzymes, organelles over the whole cell. (a) Examples of AEs (metalloenzymes (i), cyclodextrin as an example of a self‐assembled supramolecular scaffold (ii) and inorganic particles with polymeric imprinting (iii)). (b) AOs (liposomes with encapsulated enzymes (i), synthetic hollow mesoporous nanospheres (ii), polymersome with membrane pores that allow small molecules to transport across the membrane (iii) and a micelle with an active enzymatic core (iv)). (c) ACs (giant vesicle with natural organelles encapsulated (i), hydrogel bead with encapsulated liposomes with biocatalytic activity (ii) are shown schematically). Typical size ranges are indicated in the scale bar at the bottom
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Artificial cells. (a) Encapsulated catalysis: (i) Schematic presentation of SaSO2 cells co‐cultured with matrix vesicle (MV) loaded alginate beads to create spheroids; (ii) Relative total Ca2+‐content quantified after 3, 7, and 14 days of incubation of spheroids in osteoconductive media consisting of cells only (S‐C), empty alginate beads and cells (S‐ME) or MV‐loaded alginate beads and cells (S‐MMV). (b) Communication: (i) Schematic of the activator and reporter cell pair. The activator cell contains the template for T3 RNA polymerase (T3 RNAP) expression. Reporter cells undergo T3 RNAP driven expression of a fluorescent fusion protein of the tetracycline repressor TetR and sfGFP (TetR‐sfGFP) and the tet operator sequence (tetO) array plasmid to localize the TetR‐sfGFP fluorescence to the hydrogel nucleus. Micrographs show a rhodamine B fluorescence in activator membranes and TetR‐sfGFP in the hydrogel nucleus of reporter cells; (ii) The artificial quorum sensing cell contain DNA templates for T3 activation cascade and tetO array plasmid which drives tetR‐sfGFP production; (iii) Micrographs of ACs in droplets of cell‐free transcription and translation (TX‐TL) reagents with the number of cells is indicated (left panel). The enlarged regions shown in the right panel, indicated with the white box on left images, show green fluorescence after 3 h of incubation. (c) Energy transduction: (i) Overview of the AC; (ii) Schematic of the AC containing the PL and translation‐only PURE system to facilitate bacteriorhodopsin (bR) and ATP synthase subunit F0 synthesis; (iii) Confocal laser scanning microscopy images of light‐induced de novo bR‐GFP production inside GUVs; iv) Light‐driven ATP synthesis by PLs with de novo F0 subunit synthesis with wild type (awt) or mutant (amut) subunit was synthesized. Panel a reprinted with permission from Itel et al., (2018). Copyright 2018 American Chemical Society. Panel b reprinted from Niederholtmeyer et al., (2018). Copyright 2018 Springer Nature. Panel c reprinted from Berhanu et al., (2019). Copyright 2019 Springer Nature
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Artificial organelles. (a) (i) Cartoon illustrating the cytosolic placed nanoreactors loaded with glucose oxidase or β‐galacotosidase; ii) NO production in primary human macrophages exposed to β‐Gal‐NONOate. Cell mean fluorescence (CMF) originated from the interaction with 4‐amino‐5‐methyl‐amino‐2,7‐difluorofluorescein diacetate and intracellular produced NO. Data represent mean ± SD (**p < .01). (b) (i) PEG−PClgTMC polymersomes loaded with enzymatic cargo and surface functionalized with cell‐penetrating peptide (CPP); (ii) Functional analysis using primary skin fibroblasts from a healthy individual (C5120) and a patient with isolated complex I deficiency (S7‐5175). Intracellular ROS assessed by chloromethyl‐2,7‐dichlorodihydrofluorescein diacetate fluorescence intensity measurements when cells pre‐treated with polymersomes were challenged with H2O2 for 5 min. (c) (i) Illustration of RuSCNP‐enzyme co‐delivery and dual catalysis; (ii) Illustration of SCNP‐enzyme tandem reaction conducted with RuSCNP and βGal. Flow‐cytometry analysis of E. coli cells conducted with/without 8, RuSCNP, βGal, and irradiation. Panel a reprinted from Zhang, Gal, et al., (2019). Copyright 2019 the Royal Society of Chemistry. Panel b reprinted from van Oppen et al., (2018). Copyright 2018 American Chemical Society. Panel c reprinted with permission from Chen et al., (2020). Copyright 2020 American Chemical Society
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Artificial enzymes. (a) (i) Designing strategy of the ArM for the fluorescence turn‐on detection of ethylene; (ii) Fluorescence imaging of a kiwi fruit with the ArM in three selected sections during the ripening process. (b) (i) Chemical structures of the block copolymer P1 and EUK; (ii) Schematic illustration of the internalization of the artificial organelles; (iii) Cell viability of HepG2 cells loaded with artificial organelle after stressed with paraquat after 24 h. The data are expressed as mean ± SD (n = 3, *p < .05, **p < .01). (c) (i) Schematic illustration of the molecularly imprinting TMB binding pockets; (ii) Photographs showing the selectivity differences for TMB with and without treatment of molecular imprinted polymers. (d) (i) Schematic illustration of N‐PCNSs induced tumor cell destruction; (ii) TEM image of cancer cells treated with HFn‐N‐PCNSs. Red arrows indicate the location of HFn‐N‐PCNSs. Scale bar = 500 nm; (iii) Photographs showing the time‐dependent evolution and progress of human HepG2 tumor morphology after treatment with HFn‐N‐PCNSs‐3 (3 indicates a high doping level of nitrogen). Panel a reprinted with permission from Vong et al., (2019). Copyright 2019 Springer Nature. Panel b reprinted with permission from Ade et al., (2019). Copyright 2019 American Chemical Society. Panel c reprinted with permission from Zhang, et al., (2017). Copyright 2017 American Chemical Society. Panel d reprinted from Fan et al., (2018). Copyright 2018 Springer Nature
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