Methods to mechanically perturb and characterize GUV-based minimal cell models

Wubshet, Nadab; Liu, Allen

doi:10.1016/j.csbj.2022.12.025

Cited by 8 publications

(9 citation statements)

References 183 publications

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“…[25,59] Currently, more than twenty different techniques exist, giving offspring to hundreds of different protocols, ranging from relatively straightforward bulk techniques to sophisticated microfluidic assembly lines. [16,57,60,61] GUV production methods are typically classified into two main categories: swelling-based approaches, which rely on rehydrating dried lipid bilayers, and emulsion-based approaches, where lipids are initially adsorbed to water-oil interfaces and a bilayer is only formed after subsequent transfer through an oil-water interface (Figure 2).…”

Section: Overview Of Available Methods For Guv Productionmentioning

confidence: 99%

“…However, GUV membranes are naturally unstable and lack robust mechanical properties as compared to their natural counterparts, cell membranes. As reviewed in Wubshet et al, [ 57 ] GUV stability can be enhanced by membrane modulation, e.g., by tuning the lipid composition, changing bilayer asymmetry, or altering the cholesterol concentration, or by luminal modulation, e.g., by encapsulating cytoskeletal components or other, structurally relevant molecules.…”

Section: Guvs As the Container For Artificial Cells—what Are The Requ...mentioning

confidence: 99%

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Exploring Giant Unilamellar Vesicle Production for Artificial Cells — Current Challenges and Future Directions

et al. 2023

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Creating an artificial cell from the bottom up is a long‐standing challenge and, while significant progress has been made, the full realization of this goal remains elusive. Arguably, one of the biggest hurdles that researchers are facing now is the assembly of different modules of cell function inside a single container. Giant unilamellar vesicles (GUVs) have emerged as a suitable container with many methods available for their production. Well‐studied swelling‐based methods offer a wide range of lipid compositions but at the expense of limited encapsulation efficiency. Emulsion‐based methods, on the other hand, excel at encapsulation but are only effective with a limited set of membrane compositions and may entrap residual additives in the lipid bilayer. Since the ultimate artificial cell will need to comply with both specific membrane and encapsulation requirements, there is still no one‐method‐fits‐all solution for GUV formation available today. This review discusses the state of the art in different GUV production methods and their compatibility with GUV requirements and operational requirements such as reproducibility and ease of use. It concludes by identifying the most pressing issues and proposes potential avenues for future research to bring us one step closer to turning artificial cells into a reality.

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Section: Overview Of Available Methods For Guv Productionmentioning

confidence: 99%

Section: Guvs As the Container For Artificial Cells—what Are The Requ...mentioning

confidence: 99%

Exploring Giant Unilamellar Vesicle Production for Artificial Cells — Current Challenges and Future Directions

et al. 2023

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“…Cellular organelles may also appear as core/shell systems in which the interior of the organelle is the core, while the boundary, often a lipid bilayer, constitutes the shell. 16 Even solute/solvent interactions may be represented as core/shell structures, where the solute occupies a void in the solvent� considered the core�and the solvation shells act as their namesake: the shell. 9,10 In many of these scenarios, the shape of the core/shell assembly is important to the function of the particle.…”

Section: Introductionmentioning

confidence: 99%

“…Solid nanoparticles can be observed using techniques such as electron microscopy. 1,16,[20][21][22][23][24][25]28 Similarly, protein structures, and thus their shapes, may be experimentally determined using, for example, X-ray diffraction, NMR, and cryo-electron microscopy. 29−31 However, for both nanoparticles and proteins, we are limited to measurements of shape only after fully determining their structure with very little a priori understanding of the shape.…”

Section: Introductionmentioning

confidence: 99%

“…Although core/shell nanoparticles are perhaps the most obvious example of this phenomenon and have applications in many of the fields identified, − this motif is observed in many other contexts. − A surfactant layer that stabilizes many nanoparticles presents a “shell” layer around the nanoparticle “core.” Soft materials or supramolecular assemblies, such as micelles , or reverse micelles, fall into this pattern too; a micelle essentially comprises a core coated with a surfactant layer as a shell around it. Cellular organelles may also appear as core/shell systems in which the interior of the organelle is the core, while the boundary, often a lipid bilayer, constitutes the shell . Even solute/solvent interactions may be represented as core/shell structures, where the solute occupies a void in the solventconsidered the coreand the solvation shells act as their namesake: the shell. , …”

Section: Introductionmentioning

confidence: 99%

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Predicting the Geometry of Core–Shell Structures: How a Shape Changes with Constant Added Thickness

Gale,

Levinger

2024

J. Phys. Chem. B

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The core−shell assembly motif is ubiquitous in chemistry. While the most obvious examples are core/shell-type nanoparticles, many other examples exist. The shape of the core/ shell constructs is poorly understood, making it impossible to separate chemical effects from geometric effects. Here, we create a model for the core/shell construct and develop proof for how the eccentricity is expected to change as a function of the shell. We find that the addition of a constant thickness shell always creates a relatively more spherical shape for all shapes covered by our model unless the shape is already spherical or has some underlying radial symmetry. We apply this work to simulated AOT reverse micelles and demonstrate that it is remarkably successful at explaining the observed shapes of the chemical systems. We identify the three specific cases where the model breaks down and how this impacts eccentricity.

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Rearrangement of GUV‐confined actin networks in response to micropipette aspiration

Wubshet,

Young,

Liu

2024

Cytoskeleton

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Although diverse actin network architectures found inside the cell have been individually reconstituted outside of the cell, how different types of actin architectures reorganize under applied forces is not entirely understood. Recently, bottom‐up reconstitution has enabled studies where dynamic and phenotypic characteristics of various actin networks can be recreated in an isolated cell‐like environment. Here, by creating a giant unilamellar vesicle (GUV)‐based cell model encapsulating actin networks, we investigate how actin networks rearrange in response to localized stresses applied by micropipette aspiration. We reconstitute actin bundles and branched bundles in GUVs separately and mechanically perturb them. Interestingly, we find that, when aspirated, protrusive actin bundles that are otherwise randomly oriented in the GUV lumen collapse and align along the axis of the micropipette. However, when branched bundles are aspirated, the network remains intact and outside of the pipette while the GUV membrane is aspirated into the micropipette. These results reveal distinct responses in the rearrangement of actin networks in a network architecture‐dependent manner when subjected to physical forces.

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Methods to mechanically perturb and characterize GUV-based minimal cell models

Cited by 8 publications

References 183 publications

Exploring Giant Unilamellar Vesicle Production for Artificial Cells — Current Challenges and Future Directions

Exploring Giant Unilamellar Vesicle Production for Artificial Cells — Current Challenges and Future Directions

Predicting the Geometry of Core–Shell Structures: How a Shape Changes with Constant Added Thickness

Rearrangement of GUV‐confined actin networks in response to micropipette aspiration

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