Temporally controlled multistep division of DNA droplets for dynamic artificial cells

Nature Communications, Год журнала: 2024, Номер 15(1)

Опубликована: Авг. 27, 2024

Abstract Synthetic droplets mimicking bio-soft matter formed via liquid-liquid phase separation (LLPS) in living cells have recently been employed nanobiotechnology for artificial cells, molecular robotics, computing, etc. Temporally controlling the dynamics of synthetic is essential developing such bio-inspired systems because maintain their functions based on temporally controlled biomolecular reactions and assemblies. This paper reports temporal control DNA-based LLPS (DNA droplets). We demonstrate timing-controlled division DNA time-delayed triggers regulated by chemical reactions. Controlling release order multiple results multistep droplet division, i.e., pathway-controlled a reaction landscape. Finally, we apply into computing element to compare microRNA concentrations. believe that will promote design dynamic cells/molecular robots sophisticated biomedical applications.

Язык: Английский

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Reactive Brownian Dynamics of Chemically Fueled Droplets: Roles of Attraction and Deactivation Modes

The Journal of Physical Chemistry B, Год журнала: 2025, Номер 129(2), С. 626 - 636

Опубликована: Янв. 6, 2025

The self-assembly of biological membraneless organelles can be mimicked by active droplets resulting from chemically fueled microphase separation. However, how the nonequilibrium, transient structure these controlled through physicochemical input parameters is not yet well understood. In our work, a two-state chemical reaction and subsequent droplet growth decay are modeled with reactive Brownian dynamics simulation in two spatial dimensions. model, particles that activated via consumption fuel become attractive accumulate into droplets. A local-density-dependent distinction droplet's 'internal' 'external' allows for structural feedback giving further control over deactivation process. shows only external slows down stabilizes droplets, whereas internal lead to temporary encapsulation deactivated (in nonequilibrium 'core–shell' structures) where serve as an outer shell. Additionally, role hydrophobicity resembled attraction energy ε dependency formation on various investigated. For example, high finite-size crystalline while other parameter choices indicate bimodal size distributions at specific times. Similarities differences related experiments discussed.

Язык: Английский

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Size control and oscillations of active droplets in synthetic cells

Nature Communications, Год журнала: 2025, Номер 16(1)

Опубликована: Фев. 26, 2025

Abstract Oscillations in the formation and dissolution of molecular assemblies inside living cells are pivotal orchestrating various cellular functions processes. However, designing such rhythmic patterns synthetic remains a challenge. Here, we demonstrate spontaneous emergence spatio-temporal oscillations number droplets, size, their spatial distribution within cell. The coacervate-based droplets these sediment fuse at cell’s bottom. Through size control mechanism, sedimented, large shrink by expelling droplet material. expelled molecules nucleate new top cell, which grow again. These sustained converting chemical fuel into waste can continue for hundreds periods without evidence fatigue. Strikingly, period oscillation is minute’s regime tunable. design oscillating artificial organelles brings us closer to creating more life-like materials de novo life.

Язык: Английский

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Programmable Coacervate Droplets via Reaction-Coupled Liquid–Liquid Phase Separation (LLPS) and Competitive Inhibition

Journal of the American Chemical Society, Год журнала: 2025, Номер unknown

Опубликована: Март 20, 2025

Membraneless biomolecular condensates formed by liquid–liquid phase separation (LLPS) are crucial for many spatiotemporal biological functions. Designing synthetic mimics to emulate and understand LLPS is an active area of research, which has led the development coacervate droplets through elegant bioinspired designs. However, recent interest in this field shifted toward designing programmable coacervates impart control over these liquid phases. Herein, we demonstrate programming systems employing concepts competitive binding reaction-coupled assembly involving dynamic covalent bonds. Our results utilize small building blocks that follow a simple coacervation mechanism, distinguishing approach from previously reported complex coacervates, often rely on reaction-controlled generation one components. We introduce using bonds (boronate esters) chromophoric appended with terminal boronic acid groups. Upon reaction substrates (monosaccharides), form molecular structures resembling "sticker-and-spacer" designs coacervation, leading reaction-driven, temporally controlled process. The differential reactivity various monosaccharides, combined reversibility bonds, enables binding-driven growth, inhibition, dissolution process, offering new strategies reminiscent protein-induced inhibition condensates. Detailed spectroscopic probing kinetic analyses provide mechanistic insights into autocatalytic growth processes, revealing glucose-selective nature system. Finally, coupling reactions temporal pH modulation transient can be visualized confocal microscopy. anticipate will pave way novel biorelevant emergent properties.

Язык: Английский

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Spontaneous Formation of Core–Shell Microdroplets during Conventional Coacervate Phase Separation

Langmuir, Год журнала: 2025, Номер unknown

Опубликована: Март 25, 2025

We report the single-step formation and stability of protocell-like, core-shell coacervate droplets comprising a polyelectrolyte-rich shell solvent-rich vacuole core from poly(allylamine hydrochloride) (PAH) poly(acrylic acid) (PAA) system. These double emulsion (DE) coexist with single (SE) droplets, suggesting kinetic mechanism formation. use high-throughput microscopy machine learning to classify droplet morphologies across various final compositions (polyelectrolyte ratios salt concentrations) processing routes (mixing rate thermodynamic path). find that DE form preferentially over SE at wide range using slow injection mixing rate. is enhanced lower (NaCl) levels near 1:1 charge stoichiometry, showing preference for polycation excess. are stable micron scale retain their structure even after coalescence. Nevertheless, they metastable; direct observations coarsening phenomena suggest primarily stabilized by viscoelasticity high viscosity polymer-rich shell. Overall, scalable, simple process used herein offers novel produce multiphase orthogonal existing routes, which require either dropwise synthesis or tuning.

Язык: Английский

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Activity-Induced Droplet Inversion in Multicomponent Liquid–Liquid Phase Separation

Journal of Chemical Theory and Computation, Год журнала: 2025, Номер unknown

Опубликована: Март 26, 2025

Liquid–liquid phase separation (LLPS) is a vital process in forming membrane-free organelles, crucial for cell physiology and recently gaining significant attention. However, the effects of nonequilibrium factors, which are common real life, on LLPS have not been fully explored. To address this issue, we developed model involving three components (A, B, C) by integrating term into chemical potential active component B. We find changes morphology dynamics phase-separated droplets compared to their equilibrium counterparts. Remarkably, with large enough activity, B-A-C structure (B at center, surrounded A, then enveloped under conditions may change C-A-B structure. Further simulations give global picture system both passive conditions, revealing shifts boundaries unraveling effect activity different droplet structures. derived an effective free energy provide qualitative understanding our observations. Our study presents basic processes, providing insights alongside intracellular phenomena.

Язык: Английский

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Chemically active wetting

Proceedings of the National Academy of Sciences, Год журнала: 2025, Номер 122(15)

Опубликована: Апрель 9, 2025

Wetting of liquid droplets on passive surfaces is ubiquitous in our daily lives, and the governing physical laws are well understood. When become active, however, wetting remain elusive. Here, we propose chemically active as a class systems where surface due to binding process that maintained away from equilibrium. We derive corresponding nonequilibrium thermodynamic theory show fundamentally changes behavior, leading steady, states with droplet shapes reminiscent pancake or mushroom. The origin such anomalous can be explained by mapping electrostatics, pairs sinks sources correspond electrostatic dipoles along triple line. This an example more general analogy, localized chemical activity gives rise multipole field potential. underlying physics relevant for cells, droplet-forming proteins bind membranes accompanied turnover biological fuels.

Язык: Английский

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Formation of liquid shells in active droplet systems

Physical Review Research, Год журнала: 2023, Номер 5(4)

Опубликована: Дек. 14, 2023

We study a chemically active binary mixture undergoing phase separation and show that under nonequilibrium conditions, stable liquid spherical shells can form via spinodal instability in the droplet center. A single shell tends to grow until it undergoes shape beyond critical size. In an emulsion, many stationary coexist. discuss conditions which are dominant as compared regimes where droplets undergo instabilities divide.

Язык: Английский

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Membrane Transport, Molecular Machines, and Maxwell's Demon

ChemSystemsChem, Год журнала: 2024, Номер 6(3)

Опубликована: Фев. 9, 2024

Abstract The spontaneous generation of transmembrane gradients is an important fundamental research goal for artificial nanotechnology. active transport processes that give rise to such directly mirror the famous Maxwell's Demon thought experiment, where a partitions particles between two chambers generate nonequilibrium state. Despite these similarities, discussion absent in literature on membrane transport. By contrast, emergence rational design principles molecular motors can trace its intellectual roots this experiment. This perspective highlights links and machines, argues understanding implications 19 th century experiment crucial future development processes.

Язык: Английский

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Nonequilibrium Wet–Dry Cycling Acts as a Catalyst for Chemical Reactions

The Journal of Physical Chemistry B, Год журнала: 2024, Номер 128(7), С. 1724 - 1736

Опубликована: Фев. 9, 2024

Recent experimental studies suggest that wet–dry cycles and coexisting phases can each strongly alter chemical processes. The mechanisms of why to what degree processes are altered when subjected evaporation condensation unclear. To close this gap, we developed a theoretical framework for nondilute reactions subject nonequilibrium conditions condensation. We find such change the half-time product's yield by more than an order magnitude, depending on substrate–solvent interaction. show cycle frequency affects turnover system is maintained out equilibrium cycles. There exists resonance behavior in where maximal. This enables select specific reactions, suggesting potential mechanism evolution prebiotic soups at early Earth.

Язык: Английский

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