Coacervate Droplets for Synthetic Cells

Small Methods, Journal Year: 2023, Volume and Issue: 7(12)

Published: July 18, 2023

Abstract The design and construction of synthetic cells – human‐made microcompartments that mimic features living have experienced a real boom in the past decade. While many efforts been geared toward assembling membrane‐bounded compartments, coacervate droplets produced by liquid–liquid phase separation emerged as an alternative membrane‐free compartmentalization paradigm. Here, dual role cell research is discussed: encapsulated within membrane‐enclosed coacervates act surrogates membraneless organelles ubiquitously found cells; alternatively, they can be viewed crowded cytosol‐like chassis for constructing integrated cells. After introducing key concepts coacervation illustrating chemical diversity systems, their physicochemical properties resulting bioinspired functions are emphasized. Moving from suspensions free floating coacervates, two nascent roles these highlighted: organelle‐like modules templates. Building discussion on recent studies literature, potential to assemble capable multiple life‐inspired showcased. Future challenges still tackled field finally discussed.

Language: Английский

---

Liquid spherical shells are a non-equilibrium steady state of active droplets

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: Oct. 17, 2023

Liquid-liquid phase separation yields spherical droplets that eventually coarsen to one large, stable droplet governed by the principle of minimal free energy. In chemically fueled separation, formation phase-separating molecules is coupled a fuel-driven, non-equilibrium reaction cycle. It thus dissipative structures sustained continuous fuel conversion. Such are ubiquitous in biology but poorly understood as they thermodynamics. Here, we bridge gap between passive, close-to-equilibrium, and active, with separation. We observe spherical, active can undergo morphological transition into liquid, shell. demonstrate mechanism related gradients short-lived material. characterize how far out equilibrium shell state chemical power necessary sustain it. Our work suggests alternative avenues for assembling complex morphologies, which might already be exploited form membraneless organelles cells.

Language: Английский

---

Chemically Fueled Supramolecular Materials

Accounts of Materials Research, Journal Year: 2023, Volume and Issue: 4(5), P. 416 - 426

Published: April 14, 2023

In biology, the function of many molecules is regulated through nonequilibrium chemical reaction cycles. The prototypical example phosphorylation an amino acid in enzyme which induces a functional change, e.g., it folds or unfolds, assembles disassembles, binds substrate. Such does not occur spontaneously but requires phosphorylating agent with high potential (for example, adenosine triphosphate (ATP)) to be converted into molecule lower (adenosine diphosphate (ADP)). When this energy used regulate assembly, we speak chemically fueled assemblies; i.e., potential, fuel, self-assembly process. For binding guanosine (GTP) tubulin self-assembly. bound GTP hydrolyzed (GDP) upon disassembly. result dynamic assembly endowed unique characteristics, such as time-dependent behavior and ability self-heal. These intriguing, properties have inspired supramolecular chemists create similar molecular assemblies from bottom up. While examples been designed, they remain scarce partly because cycles are rare often complex. Thus, recently developed carbodiimide-driven cycle that versatile easy use, quantitatively understood, suffer side reactions. cycle, carboxylate precursor reacts carbodiimide form activated species like anhydride ester. state water thereby reverts its state; deactivated. Effectively, catalyzes carbodiimides' conversion waste forms transient state. We designed building blocks range materials at expense fuel. simplicity versatility democratized popularized field assemblies. Account, describe what "learned" on our way. introduce exemplified by biological design cycle. Using group others, offer rules for block's structure strategies desired morphology materials. discussed morphologies include fibers, colloids, crystals, oil- coacervate-based droplets. then demonstrate how these material Besides, discuss concept reciprocal coupling exerts feedback also mechanisms. Finally, close Account discussion outlook field. This aims provide fundamental understanding facilitate further progress toward conceptually new

Language: Английский

---

Phase Transitions in Chemically Fueled, Multiphase Complex Coacervate Droplets

Angewandte Chemie International Edition, Journal Year: 2022, Volume and Issue: 61(46)

Published: Sept. 6, 2022

Abstract Membraneless organelles are droplets in the cytosol that regulated by chemical reactions. Increasing studies suggest they internally organized. However, how these subcompartments remains elusive. Herein, we describe a complex coacervate‐based model composed of two polyanions and short peptide. With reaction cycle, control affinity peptide for polyelectrolytes leading to distinct regimes inside phase diagram. We study transitions from one regime another identify new can only occur under kinetic control. Finally, show cycle controls liquidity offering insights into active processes cells play an important role tuning liquid state membraneless organelles. Our work demonstrates not thermodynamic properties but also kinetics should be considered organization multiple phases droplets.

Language: Английский

---

Hydrophilic polymers: Current trends and visions for the future

Progress in Polymer Science, Journal Year: 2023, Volume and Issue: 147, P. 101753 - 101753

Published: Oct. 18, 2023

Hydrophilic polymers are a major class of in polymer science. They found broad range applications from superabsorbers to drug-delivery. In recent years, plethora impactful developments hydrophilic have been reported. The present review gives an overview over these with focus on frequently studied types, aqueous multi-phase systems, block copolymer self-assembly and particles. We cover fundamental work concepts but also high relevance for application. Finally, we give outlook towards current challenges future the field. further development is great importance will significant impact biomedicine every-day life.

Language: Английский

---

Non-enzymatic oligonucleotide ligation in coacervate protocells sustains compartment-content coupling

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: May 9, 2023

Abstract Modern cells are complex chemical compartments tightly regulated by an underlying DNA-encoded program. Achieving a form of coupling between molecular content, reactions, and chassis in synthetic represents key step to the assembly evolvable protocells but remains challenging. Here, we design coacervate droplets that promote non-enzymatic oligonucleotide polymerization restructure as result reaction dynamics. More specifically, rationally exploit complexation end-reactive oligonucleotides able stack into long physical polymers cationic azobenzene photoswitch produce three different phases—soft solids, liquid crystalline or isotropic coacervates droplets—each them having impact on efficiency. Dynamical modulation dissolution via trans - cis photo-isomerization is used demonstrate cycles light-actuated ligation. Remarkably, changes population polynucleotides during induce phase transitions due length-based DNA self-sorting multiphase coacervates. Overall, combining tight reaction-structure environmental responsiveness, our reactive provide general route synthesis pave way emergence primitive compartment-content membrane-free protocells.

Language: Английский

---

Suppressing catalyst poisoning in the carbodiimide-fueled reaction cycle

Chemical Science, Journal Year: 2023, Volume and Issue: 14(44), P. 12653 - 12660

Published: Jan. 1, 2023

In biology, cells regulate the function of molecules using catalytic reaction cycles that convert reagents with high chemical potential (fuel) to waste molecules. Inspired by synthetic analogs such have been devised, and a widely used cycle uses carboxylates as catalysts accelerate hydration carbodiimides. The is versatile easy use, so it applied motors, pumps, self-assembly, phase separation. However, suffers from side reactions, especially formation N-acylurea. cycles, reactions are disastrous they decrease fuel's efficiency and, more importantly, destroy molecular machinery or assembling Therefore, this work tested how suppress N-acylurea screening precursor concentration, its structure, carbodiimide additives, temperature, pH. It turned out combination low pH, 10% pyridine fraction fuel could significantly product keep highly effective successful assembly. We anticipate our will provide guidelines for carbodiimide-fueled choose optimal conditions.

Language: Английский

---

Multicompartmental coacervate-based protocell by spontaneous droplet evaporation

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: Feb. 6, 2024

Abstract Hierarchical compartmentalization, a hallmark of both primitive and modern cells, enables the concentration isolation biomolecules, facilitates spatial organization biochemical reactions. Coacervate-based compartments can sequester recruit large variety molecules, making it an attractive protocell model. In this work, we report spontaneous formation core-shell cell-sized coacervate-based driven by evaporation sessile droplet on thin-oil-coated substrate. Our analysis reveals that such far-from-equilibrium architectures arise from multiple, coupled segregative associative liquid-liquid phase separation, are stabilized stagnation points within evaporating droplet. The results convective capillary flows induced maximum rate at liquid-liquid-air contact line. This work provides valuable insights into maintenance hierarchical under non-equilibrium conditions, offering glimpse real-life scenario.

Language: Английский

---

Chemotactic Interactions Drive Migration of Membraneless Active Droplets

Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(23), P. 15965 - 15976

Published: April 15, 2024

In nature, chemotactic interactions are ubiquitous and play a critical role in driving the collective behavior of living organisms. Reproducing these vitro is still paramount challenge due to complexity mimicking controlling cellular features, such as tangled metabolic networks, cytosolic macromolecular crowding, migration, on microorganism size scale. Here, we generate enzymatically active cell-sized droplets able move freely, by following chemical gradient, interact with surrounding manner. The enzyme within generates pH gradient that extends outside edge droplets. We discovered external triggers droplet migration controls its directionality, which selectively toward neighboring Hence, changing activity inside droplet, tuned speed. Furthermore, showed cellular-like features can facilitate reconstitution simple linear protometabolic pathway increase final reaction product generation. Our work suggests stable membraneless reproduce complex biological phenomena, opening new perspectives bioinspired materials synthetic biology tools.

Language: Английский

---

Synthetic‐Cell‐Based Multi‐Compartmentalized Hierarchical Systems

Small Methods, Journal Year: 2023, Volume and Issue: 7(12)

Published: April 17, 2023

Abstract In the extant lifeforms, self‐sustaining behaviors refer to various well‐organized biochemical reactions in spatial confinement, which rely on compartmentalization integrate and coordinate molecularly crowded intracellular environment complicated reaction networks living/synthetic cells. Therefore, biological phenomenon of has become an essential theme field synthetic cell engineering. Recent progress state‐of‐the‐art cells indicated that multi‐compartmentalized should be developed obtain more advanced structures functions. Herein, two ways developing hierarchical systems, namely interior (organelles) integration communities (synthetic tissues), are summarized. Examples provided for different construction strategies employed above‐mentioned engineering ways, including spontaneous vesicles, host–guest nesting, phase separation mediated multiphase, adhesion‐mediated assembly, programmed arrays, 3D printing. Apart from exhibiting functions, also applied as biomimetic materials. Finally, key challenges future directions regarding development systems summarized; these expected lay foundation creation a “living” well provide larger platform new materials future.

Language: Английский

---