Giant vesicles: a powerful tool to reconstruct bacterial division assemblies in cell‐like compartments

Jiménez, Mercedes; Martos, Ariadna; Cabré, Elisa J.; Raso, Ana; Rivas, Germán

doi:10.1111/1462-2920.12214

Cited by 20 publications

(17 citation statements)

References 71 publications

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“…They are composed by freestanding phospholipid bilayers and, due to their large size (from 5 to 100 μm of diameter), can be analyzed by means of optical microscopy. GUVs can be formed from different lipid mixtures, including bacterial membranes [13] and the available strategies for their manipulation allow their use as cell-sized biological reactors and minimal bacterial machineries [21].…”

Section: Reconstitution Of Cytotoxic Bacterial Protein Assemblies In mentioning

confidence: 99%

Reconstruction of Cytotoxic Bacterial Protein Assemblies in Lipid Vesicles

Fernández

Rivas

Giraldo

et al. 2017

Advances in Biomembranes and Lipid Self-Assembly

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Biochemistry is at the core of constructive bottom-up approaches in synthetic biology aiming to generate a minimal cell. Any sort of cellular organization depends on compartmentalization of macromolecular assemblies and machineries within lipid, or amphiphilic lipid-like, vesicles. Following the increasing interest in the use of liposomes as cytomimetic devices, here we discuss our achievements toward using liposomes, especially giant unilamellar vesicles, as containers to reconstitute the amyloidogenic aggregation of the cytotoxic prionoid (prion-like protein) RepA-WH1. We address relevant issues as the importance of lipid composition and the effect or cofactors on assembly, and the feasibility of achieving protein expression within vesicles.

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Section: Reconstitution Of Cytotoxic Bacterial Protein Assemblies In mentioning

confidence: 99%

Reconstruction of Cytotoxic Bacterial Protein Assemblies in Lipid Vesicles

Fernández

Rivas

Giraldo

et al. 2017

Advances in Biomembranes and Lipid Self-Assembly

Self Cite

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“…7, 8, 17 Early versions of the technique pioneered the centrifugation of polydisperse reverse emulsions through an oil-water interface. 37, 38 Droplet polydispersity and lack of control of how droplets cross the interface can lead to poor yield.…”

Section: Introductionmentioning

confidence: 99%

cDICE method produces giant lipid vesicles under physiological conditions of charged lipids and ionic solutions

2016

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Giant unilamellar vesicles are a powerful and common tool employed in biophysical studies of lipid membranes. Here we evaluate a recently introduced method of vesicle formation, “continuous droplet interface crossing encapsulation” (cDICE). This method produces mondisperse giant unilamellar vesicles of controlled sizes and high encapsulation efficiencies, using readily available instrumentation. We find that mixtures of phospholipids within vesicle membranes produced by cDICE undergo phase separation at the same characteristic temperatures as lipids in vesicles formed by a complementary technique. We find that the cDICE method is effective both when vesicles are produced from charged lipids and when the surrounding buffer contains a high concentration of salt. A shortcoming of the technique is that cholesterol is not substantially incorporated into vesicle membranes.

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“…[17a] Synthetic mimetics of cell membranes are,f or example,g iant unilamellar vesicles (GUVs), which can be formed from predefined phospholipid constituents. [182] Synthetic vesicles are used as model systems to investigate membrane protein functions,but they also serve as compartments for the bottom-up construction of minimal cellular systems.S uch biomimetic systems (e.g.G UVs and others) that contain functional parts or systems have the capacity to produce energy, [183] to run atranscriptional oscillator, [184] or to perform biochemical reactions and so represent small celllike bioreactors. [185] An example of reconstituting ac ellular phenomenon is based on the MinCDE protein system from E. coli,which has been studied in cell-free environments to identify the minimal requirements for maintaining its functionality.T he presence of the two purified proteins MinD and MinE, af lat mem-brane,a nd ATPw as sufficient to initiate the dynamic selforganization of the proteins (see Figure 5D)and result in the formation of surface waves.…”

Section: Reconstitution Of Minimal Synthetic Systemsmentioning

confidence: 99%

Synthetic Biology—The Synthesis of Biology

2017

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Synthetic biology concerns the engineering of man-made living biomachines from standardized components that can perform predefined functions in a (self-)controlled manner. Different research strategies and interdisciplinary efforts are pursued to implement engineering principles to biology. The "top-down" strategy exploits nature's incredible diversity of existing, natural parts to construct synthetic compositions of genetic, metabolic, or signaling networks with predictable and controllable properties. This mainly application-driven approach results in living factories that produce drugs, biofuels, biomaterials, and fine chemicals, and results in living pills that are based on engineered cells with the capacity to autonomously detect and treat disease states in vivo. In contrast, the "bottom-up" strategy seeks to be independent of existing living systems by designing biological systems from scratch and synthesizing artificial biological entities not found in nature. This more knowledge-driven approach investigates the reconstruction of minimal biological systems that are capable of performing basic biological phenomena, such as self-organization, self-replication, and self-sustainability. Moreover, the syntheses of artificial biological units, such as synthetic nucleotides or amino acids, and their implementation into polymers inside living cells currently set the boundaries between natural and artificial biological systems. In particular, the in vitro design, synthesis, and transfer of complete genomes into host cells point to the future of synthetic biology: the creation of designer cells with tailored desirable properties for biomedicine and biotechnology.

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Giant vesicles: a powerful tool to reconstruct bacterial division assemblies in cell‐like compartments

Cited by 20 publications

References 71 publications

Reconstruction of Cytotoxic Bacterial Protein Assemblies in Lipid Vesicles

Reconstruction of Cytotoxic Bacterial Protein Assemblies in Lipid Vesicles

cDICE method produces giant lipid vesicles under physiological conditions of charged lipids and ionic solutions

Synthetic Biology—The Synthesis of Biology

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