Functional Modules of Minimal Cell Division for Synthetic Biology

Hürtgen, Daniel; Härtel, Tobias; Murray, Séan; Sourjik, Victor; Schwille, Petra

doi:10.1002/adbi.201800315

Cited by 16 publications

(11 citation statements)

References 120 publications

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“…Although aspects of our study were previously addressed individually, such as encapsulation of actin bundles, actin binding to the inner membrane leaflet of a vesicle, or encapsulation of contractile actomyosin networks in vesicles, until now it proved too experimentally challenging to reproducibly combine these within one experimental system. Our results provide a high-yield approach, returning reproducible and quantifiable results, that brings us that much closer to the ultimate goal of being able to quantitatively design and experimentally achieve full division of a synthetic membrane compartment, and thus, to the self-reproduction of artificial cells, a persistent goal in bottom-up biology [41][42][43][44] .…”

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confidence: 80%

Reconstitution of contractile actomyosin rings in vesicles

et al. 2021

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One of the grand challenges of bottom-up synthetic biology is the development of minimal machineries for cell division. The mechanical transformation of large-scale compartments, such as Giant Unilamellar Vesicles (GUVs), requires the geometry-specific coordination of active elements, several orders of magnitude larger than the molecular scale. Of all cytoskeletal structures, large-scale actomyosin rings appear to be the most promising cellular elements to accomplish this task. Here, we have adopted advanced encapsulation methods to study bundled actin filaments in GUVs and compare our results with theoretical modeling. By changing few key parameters, actin polymerization can be differentiated to resemble various types of networks in living cells. Importantly, we find membrane binding to be crucial for the robust condensation into a single actin ring in spherical vesicles, as predicted by theoretical considerations. Upon force generation by ATP-driven myosin motors, these ring-like actin structures contract and locally constrict the vesicle, forming furrow-like deformations. On the other hand, cortex-like actin networks are shown to induce and stabilize deformations from spherical shapes.

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confidence: 80%

Reconstitution of contractile actomyosin rings in vesicles

et al. 2021

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“…Out of the different cytomotive filament‐based systems of bacterial plasmid segregation, we used the best‐studied R1/ParMRC machinery . DNA segregation by this system was reconstituted by using parC ‐coated beads that, similar to a previous study, induced ParR‐dependent nucleation of ParM filaments.…”

Section: Discussionmentioning

confidence: 99%

“…Eukaryotes use dynamic spindles consisting of microtubules that rapidly reorganize by means of polymerization and depolymerization and serve as tracks for molecular motors for chromosome transport . In contrast, prokaryotes have evolved a more diverse range of DNA segregation mechanisms, in which Walker‐A ATPases of the ParA‐type commonly partition chromosomal origins of replication …”

Section: Introductionmentioning

confidence: 99%

Reconstitution and Coupling of DNA Replication and Segregation in a Biomimetic System

et al. 2019

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A biomimetic system capable of replication and segregation of genetic material constitutes an essential component for the future design of a minimal synthetic cell. Here we have used the simple T7 bacteriophage system and the plasmid‐derived ParMRC system to establish in vitro DNA replication and DNA segregation, respectively. These processes were incorporated into biomimetic compartments providing an enclosed reaction space. The functional lifetime of the encapsulated segregation system could be prolonged by equipping it with ATP‐regenerating and oxygen‐scavenging systems. Finally, we showed that DNA replication and segregation processes could be coupled in vitro by using condensed DNA nanoparticles resulting from DNA replication. ParM spindles extended over tens of micrometers and could thus be used for segregation in compartments that are significantly longer than bacterial cell size. Overall, this work demonstrates the successful bottom‐up assembly and coupling of molecular machines that mediate replication and segregation, thus providing an important step towards the development of a fully functional minimal cell.

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“…36 This work brings us much closer to our goal of being able to quantitatively design and experimentally achieve full division of a synthetic membrane compartment, and thus, to the self-reproduction of artificial cells, a persistent goal in bottom-up biology. 37-40…”

Section: Introductionmentioning

confidence: 99%

Reconstitution of contractile actomyosin rings in vesicles

Litschel

Kelley

Holz

et al. 2020

Preprint

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AbstractOne of the grand challenges of bottom-up synthetic biology is the development of minimal machineries for cell division. The mechanical transformation of large-scale compartments, such as Giant Unilamellar Vesicles (GUVs), requires the geometry-specific coordination of active elements, several orders of magnitude larger than the molecular scale. Of all cytoskeletal structures, large-scale actomyosin rings appear to be the most promising cellular elements to accomplish this task. Here, we have adopted advanced encapsulation methods to study bundled actin filaments in GUVs and compare our results with theoretical modeling. By changing few key parameters, actin polymerization can be differentiated to resemble various types of networks in living cells. Importantly, we find membrane binding to be crucial for the robust condensation into a single actin ring in spherical vesicles, as predicted by theoretical considerations. Upon force generation by ATP-driven myosin motors, these ring-like actin structures contract and locally constrict the vesicle, forming furrow-like deformations. On the other hand, cortex-like actin networks are shown to induce and stabilize deformations from spherical shapes.

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Functional Modules of Minimal Cell Division for Synthetic Biology

Abstract: of these details contributes to the applicability of functional modules for synthetic biology. Hence, this review focuses on the recent research on cell division and how the involved systems could be used in vitro to build minimal biomimetic systems, such as a minimal cell.

Cited by 16 publications

References 120 publications

Reconstitution of contractile actomyosin rings in vesicles

Reconstitution of contractile actomyosin rings in vesicles

Reconstitution and Coupling of DNA Replication and Segregation in a Biomimetic System

Reconstitution of contractile actomyosin rings in vesicles

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