Branched actin cortices reconstituted in vesicles sense membrane curvature

Baldauf, Lucia; Frey, Felix J.; Perez, Marcos Arribas; Idema, Timon; Koenderink, Gijsje H.

doi:10.1016/j.bpj.2023.02.018

Cited by 18 publications

(17 citation statements)

References 87 publications

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“…In addition, several other techniques that offer enhanced control of the inverted emulsion process have been developed. One example is continuous droplet interface crossing encapsulation (cDICE), [88,101] and its recent adaptation termed emulsion cDICE (or eDICE) [102][103][104] which have been successfully implemented for actin cytoskeleton reconstitution experiments by various labs. [88,101,[103][104][105][106][107][108] Other successful examples are the shaking method, which uses dropletstabilized GUVs as intermediates, [109] and droplet-shooting centrifugal formation (DSCF), which makes use of a 3D-printed microcapillary.…”

Section: Overview Of Available Methods For Guv Productionmentioning

confidence: 99%

“…One example is continuous droplet interface crossing encapsulation (cDICE), [88,101] and its recent adaptation termed emulsion cDICE (or eDICE) [102][103][104] which have been successfully implemented for actin cytoskeleton reconstitution experiments by various labs. [88,101,[103][104][105][106][107][108] Other successful examples are the shaking method, which uses dropletstabilized GUVs as intermediates, [109] and droplet-shooting centrifugal formation (DSCF), which makes use of a 3D-printed microcapillary. [110] While other techniques such as microfluidic jetting [111] have seen some success [112][113][114] they have not been widely adopted and further studies are needed to investigate the physicochemical properties of the resulting GUVs.…”

Section: Overview Of Available Methods For Guv Productionmentioning

confidence: 99%

“…In contrast, cDICE and its improved variant eDICE have shown compatibility with encapsulating a wide variety of biological systems. [88,103,104] Achieving successful GUV formation under the conditions needed for a given biological system of interest requires a thorough understanding of the chemical and physical properties of each system, as well as their impact on GUV formation and the encapsulation process. However, few studies have been con-ducted on the effects of different protein and protein systems on GUV formation mechanisms.…”

Section: Cross-compatibility Of Guv Production Methods and Different ...mentioning

confidence: 99%

“…[ 88 ] The more recent adaptation of cDICE, eDICE, refrains from using a capillary and results in similar GUV size distributions as cDICE. [ 104 ] This indicates an inherent size selectivity in these methods beyond that induced by droplet formation at a capillary orifice, predominantly yielding GUVs with a diameter around 10 µm. By contrast, swelling‐based methods typically result in an even more heterogeneous GUV size distribution.…”

Section: Strengths and Limitations Of Current Methods: A Comparative ...mentioning

confidence: 99%

“…[171] As an alternative to using sugars, density gradient medium OptiPrep has been successfully used in cDICE and eDICE. [88,89,[102][103][104]108,160,161] In the case of OLA, glycerol is required in both the inner and outer solution, along with the nonionic triblock copolymer surfactant Poloxamer 188 (P188) in the outer solution. It is important to note that these additional additives, especially at the high concentrations typically used, will affect protein and membrane properties during the experiments.…”

Section: Encapsulation Of Complex Solute Mixtures In Physiological Bu...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: Overview Of Available Methods For Guv Productionmentioning

confidence: 99%

Section: Cross-compatibility Of Guv Production Methods and Different ...mentioning

confidence: 99%

Section: Strengths and Limitations Of Current Methods: A Comparative ...mentioning

confidence: 99%

Section: Encapsulation Of Complex Solute Mixtures In Physiological Bu...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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Benefits and challenges of reconstituting the actin cortex

Waechtler,

Jayasankar,

Morin

et al. 2024

Cytoskeleton

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The cell's ability to change shape is a central feature in many cellular processes, including cytokinesis, motility, migration, and tissue formation. The cell constructs a network of contractile proteins underneath the cell membrane to form the cortex, and the reorganization of these components directly contributes to cellular shape changes. The desire to mimic these cell shape changes to aid in the creation of a synthetic cell has been increasing. Therefore, membrane‐based reconstitution experiments have flourished, furthering our understanding of the minimal components the cell uses throughout these processes. Although biochemical approaches increased our understanding of actin, myosin II, and actin‐associated proteins, using membrane‐based reconstituted systems has further expanded our understanding of actin structures and functions because membrane–cortex interactions can be analyzed. In this review, we highlight the recent developments in membrane‐based reconstitution techniques. We examine the current findings on the minimal components needed to recapitulate distinct actin structures and functions and how they relate to the cortex's impact on cellular mechanical properties. We also explore how co‐processing of computational models with wet‐lab experiments enhances our understanding of these properties. Finally, we emphasize the benefits and challenges inherent to membrane‐based, reconstitution assays, ranging from the advantage of precise control over the system to the difficulty of integrating these findings into the complex cellular environment.

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Membranes in focus

Sezgin¹,

Levental²

2023

Biophysical Journal

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Branched actin cortices reconstituted in vesicles sense membrane curvature

Cited by 18 publications

References 87 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

Benefits and challenges of reconstituting the actin cortex

Membranes in focus

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