In Vitro Transcription–Translation in an Artificial Biomolecular Condensate

ACS Synthetic Biology, Journal Year: 2023, Volume and Issue: 12(7), P. 2004 - 2014

Published: June 21, 2023

Biomolecular condensates are a promising platform for synthetic cell formation and constitute potential missing link between the chemical cellular stage of origins life. However, it has proven challenging to integrate complex reaction networks into biomolecular condensates, such as cell-free in vitro transcription–translation (IVTT) system. Integrating IVTT successfully is one precondition condensation-based formation. Moreover, would provide proof concept that principle compatible with central dogma, hallmarks Here, we have systemically investigated compatibility eight different (bio)molecular incorporation. Of these candidates, found green fluorescent protein-labeled, intrinsically disordered cationic protein (GFP-K72) single-stranded DNA (ssDNA) can form up μM expression. This shows indeed networks, confirming their use platforms hinting at possible role origin

Language: Английский

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Toward Artificial Cell‐Mediated Tissue Engineering: A New Perspective

Advanced Biology, Journal Year: 2023, Volume and Issue: 7(12)

Published: Aug. 10, 2023

Abstract The fast‐growing pace of regenerative medicine research has allowed the development a range novel approaches to tissue engineering applications. Until recently, main points interest in majority studies have been combine different materials control cellular behavior and use techniques optimize formation, from 3‐D bioprinting situ regeneration. However, with increase understanding fundamentals organization, development, regeneration, also come realization that for next step engineering, higher level spatiotemporal on cell‐matrix interactions is required. It proposed combination artificial cell could provide route toward over complex development. By equipping cells underlying mechanisms functions, such as communication mechanisms, migration behavior, or coherent depending surrounding matrix properties, they can be applied instructing native into desired differentiation at resolution not attained traditional materials.

Language: Английский

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Nucleic acid liquids

Reports on Progress in Physics, Journal Year: 2024, Volume and Issue: 87(6), P. 066601 - 066601

Published: May 2, 2024

The confluence of recent discoveries the roles biomolecular liquids in living systems and modern abilities to precisely synthesize modify nucleic acids (NAs) has led a surge interest liquid phases NAs. These can be formed primarily from NAs, as driven by base-pairing interactions, or electrostatic combination (coacervation) negatively charged NAs positively molecules. Generally, use sequence-engineered provides means tune microsopic particle properties, thus imbue specific, customizable behaviors into resulting liquids. In this way, researchers have used NA tackle fundamental problems physics finite valence soft materials, create with novel structured and/or multi-functional properties. Here, we review growing field, discussing theoretical background phase separation, quantitative understanding material broad array functional demonstrations these materials. We close few comments remaining open questions challenges field.

Language: Английский

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Peptide-Based Biomimetic Condensates via Liquid–Liquid Phase Separation as Biomedical Delivery Vehicles

Biomacromolecules, Journal Year: 2024, Volume and Issue: 25(9), P. 5468 - 5488

Published: Aug. 23, 2024

Biomolecular condensates are dynamic liquid droplets through intracellular liquid-liquid phase separation that function as membraneless organelles, which highly involved in various complex cellular processes and functions. Artificial analogs formed via similar pathways can be integrated with biological complexity advanced functions have received tremendous research interest the field of synthetic biology. The coacervate droplet-based compartments partition concentrate a wide range solutes, regarded attractive candidates for mimicking phase-separation behaviors biophysical features biomolecular condensates. use peptide-based materials phase-separating components has advantages such diversity amino acid residues customized sequence design, allows programming their physicochemical properties resulting compartments. In this Perspective, we highlight recent advancements design construction biomimicry from peptides relevant to protein, specific reference molecular self-assembly separation, biorelated applications, envisage emerging biomedical delivery vehicles.

Language: Английский

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Programmable Enzymatic Reaction Network in Artificial Cell‐Like Polymersomes

Advanced Science, Journal Year: 2024, Volume and Issue: 11(24)

Published: April 16, 2024

Abstract The ability to precisely control in vitro enzymatic reactions synthetic cells plays a crucial role the bottom‐up design of artificial cell models that can recapitulate key cellular features and functions such as metabolism. However, integration has been limited bulk or microfluidic emulsions without membrane, lacking more sophisticated higher‐order communities for reconstituting spatiotemporal biological information at multiple length scales. Herein, droplet microfluidics is utilized synthesize cell‐like polymersomes with distinct molecular permeability driven by external signals fuels. presence competing reverse reaction depletes active substrates shown enable demonstration fuel‐driven formation sub‐microcompartments within well realization out‐of‐equilibrium systems. In addition, different characteristics polymersome membranes are exploited successfully construct programmable network mimics communication heterogeneous community through selective transport.

Language: Английский

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Phase-separated droplets can direct the kinetics of chemical reactions including polymerization, self-replication and oscillating networks

Published: April 23, 2024

Phase-separated compartments can localize (bio)chemical reactions and influence their kinetics. They are believed to play an important role both in extant life the form of biomolecular condensates at origins as coacervate protocells. However, experimentally testing coacervates on different is practically impossible. We therefore use a numerical model explore effect phase-separated droplets kinetics outcome chemical reaction systems, where we vary volume partitioning reactants. find that rate bimolecular has optimal dilute/coacervate phase ratio for given reactant partitioning. Furthermore, accelerate polymerization self-replication lead formation longer polymers. Lastly, ‘rescue’ oscillating networks concentration regimes sustained oscillations do not occur single-phase system. Our results indicate direct wide range impact fundamental aspects such yield, pathway selection, product length emergent functions. This may have far-reaching implications life, synthetic cells fate function biological condensates.

Language: Английский

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Visualizing Formation and Dynamics of a Three-Dimensional Sponge-like Network of a Coacervate in Real Time

Journal of the American Chemical Society, Journal Year: 2023, Volume and Issue: 145(33), P. 18316 - 18328

Published: Aug. 10, 2023

Coacervates, which are formed by liquid-liquid phase separation, have been extensively explored as models for synthetic cells and membraneless organelles, so their in-depth structural analysis is crucial. However, both the inner structure dynamics formation mechanism of coacervates remain elusive. Herein, we demonstrate real-time confocal observation a three-dimensional sponge-like network in dipeptide-based coacervate. In situ generation dipeptide allowed us to capture emergence via unprecedented membrane folding vesicle-shaped intermediates. We also visualized dynamic fluctuation network, including reversible engagement/disengagement cross-links stochastic kissing event. Photoinduced transient multiphase coacervate was achieved with thermally responsive transition. Our findings expand fundamental understanding provide opportunities manipulate physicochemical properties engineering potential applications development artificial life-like material fabrication.

Language: Английский

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Multispecies autocatalytic RNA reaction networks in coacervates

Communications Chemistry, Journal Year: 2023, Volume and Issue: 6(1)

Published: May 8, 2023

Robust localization of self-reproducing autocatalytic chemistries is a key step in the realization heritable and evolvable chemical systems. While reaction networks already possess attributes such as self-reproduction evolvability, localizing functional multispecies within complex primitive phases, coacervates, has remained unexplored. Here, we show Azoarcus ribozyme system charge-rich coacervates where catalytic ribozymes are produced by assembly constituent smaller RNA fragments. We systematically demonstrate active phase-separated coacervates-both micron-sized droplets well coalesced macrophase, underscoring facility complex, phase to support these reactions multiple configurations. By constructing networks, that newly assembled molecules active, participating both self- cross-catalysis coacervates. Finally, due differential molecular transport, compartments endow robustness composition collectively against external perturbations. Altogether, our results establish formation which turn render transient network composition.

Language: Английский

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From Single‐Compartment Artificial Cells to Tissue‐Like Materials

Advanced Materials Technologies, Journal Year: 2024, Volume and Issue: 9(18)

Published: May 1, 2024

Abstract Designing and assembling artificial cells (ACs) is a core direction in bottom‐up synthetic biology. Here, the advancements past 3 years engineering ACs with focus on compartmentalization surface modifications aim for their integration semi‐synthetic tissue are outlined. Compartmentalization vesicles, coacervates hydrogels discussed encapsulated catalysis or cytoskeleton formation including use of components mammalian to increase ACs’ complexity. Following on, modification reviewed due its relevance when into goal. Finally, interaction cellular communication fabrication toward therapeutic opportunities outlined, before short perspective provided.

Language: Английский

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Phase‐separated Droplets Can Direct the Kinetics of Chemical Reactions Including Polymerization, Self‐replication and Oscillating Networks

ChemSystemsChem, Journal Year: 2024, Volume and Issue: 7(1)

Published: Aug. 22, 2024

Abstract Phase‐separated compartments can localize (bio)chemical reactions and influence their kinetics. They are believed to play an important role both in extant life the form of biomolecular condensates at origins as coacervate protocells. However, experimentally testing coacervates on different is challenging time‐consuming. We therefore use a numerical model explore effect phase‐separated droplets kinetics outcome chemical reaction systems, where we vary volume partitioning reactants. find that rate bimolecular has optimal dilute/coacervate phase ratio for given reactant partitioning. Furthermore, accelerate polymerization self‐replication lead formation longer polymers. Lastly, ‘rescue’ oscillating networks concentration regimes sustained oscillations do not occur single‐phase system. Our results indicate direct wide range impact fundamental aspects such yield, pathway selection, product length emergent functions. This may have far‐reaching implications life, synthetic cells fate function biological condensates.

Language: Английский

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