Optical Control of a Biological Reaction–Diffusion System

Glock, Philipp; Broichhagen, Johannes; Kretschmer, Simon; Blumhardt, Philipp; Mücksch, Jonas; Trauner, Dirk; Schwille, Petra

doi:10.1002/anie.201712002

Cited by 28 publications

(45 citation statements)

References 25 publications

(27 reference statements)

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“…When MinD and MinE are reconstituted on a flat supported membrane, topped by a uniform buffer, these proteins self-organize into traveling waves via an ATP-driven reaction-diffusion mechanism [74]. Such simplified flat membrane systems have been used extensively by us and others to investigate the effects of lipid and buffer composition, as well as mutations in MinD and MinE, on the formation and properties of Min patterns [75–79], and to achieve external (photo-)control over self-organization [80]. Moreover, additional division-related proteins, including MinC, FtsZ and ZipA variants, have been added to the reconstituted MinDE patterns [81–83].…”

Section: Synthetic Cell Division Via Reconstitution Of E Coli Divisomentioning

confidence: 99%

Synthetic cell division via membrane-transforming molecular assemblies

et al. 2019

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Reproduction, i.e. the ability to produce new individuals from a parent organism, is a hallmark of living matter. Even the simplest forms of reproduction require cell division: attempts to create a designer cell therefore should include a synthetic cell division machinery. In this review, we will illustrate how nature solves this task, describing membrane remodelling processes in general and focusing on bacterial cell division in particular. We discuss recent progress made in their in vitro reconstitution, identify open challenges, and suggest how purely synthetic building blocks could provide an additional and attractive route to creating artificial cell division machineries.

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Section: Synthetic Cell Division Via Reconstitution Of E Coli Divisomentioning

confidence: 99%

Synthetic cell division via membrane-transforming molecular assemblies

et al. 2019

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“…Protein expression Transform E. coli BL21 (DE3) pLysS with the respective plasmid for expression of MinD 6 , EGFP-MinD 29 , mRuby3-MinD 24 , MinE 6 or MinC 30 . For plasmid maps, please see supplementary information.…”

Section: Protocolmentioning

confidence: 99%

“…We also employed this assay to investigate how defined mutations in MinE affect pattern formation of the system 20 . Furthermore, the same basic assay format has been employed to investigate how pattern formation can be controlled by light, introducing an azobenzene-crosslinked MinE peptide into the assay, and imaging with TIRF microscopy 24 .…”

Section: Introductionmentioning

confidence: 99%

<em>In Vitro</em> Reconstitution of Self-Organizing Protein Patterns on Supported Lipid Bilayers

Ramm

Glock

Schwille

2018

JoVE

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Many aspects of the fundamental spatiotemporal organization of cells are governed by reaction-diffusion type systems. In vitro reconstitution of such systems allows for detailed studies of their underlying mechanisms which would not be feasible in vivo. Here, we provide a protocol for the in vitro reconstitution of the MinCDE system of Escherichia coli, which positions the cell division septum in the cell middle. The assay is designed to supply only the components necessary for self-organization, namely a membrane, the two proteins MinD and MinE and energy in the form of ATP. We therefore fabricate an open reaction chamber on a coverslip, on which a supported lipid bilayer is formed. The open design of the chamber allows for optimal preparation of the lipid bilayer and controlled manipulation of the bulk content. The two proteins, MinD and MinE, as well as ATP, are then added into the bulk volume above the membrane. Imaging is possible by many optical microscopies, as the design supports confocal, wide-field and TIRF microscopy alike. In a variation of the protocol, the lipid bilayer is formed on a patterned support, on cell-shaped PDMS microstructures, instead of glass. Lowering the bulk solution to the rim of these compartments encloses the reaction in a smaller compartment and provides boundaries that allow mimicking of in vivo oscillatory behavior. Taken together, we describe protocols to reconstitute the MinCDE system both with and without spatial confinement, allowing researchers to precisely control all aspects influencing pattern formation, such as concentration ranges and addition of other factors or proteins, and to systematically increase system complexity in a relatively simple experimental setup.

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“…19 Furthermore, having control over properties of light allows precise control over release or activation of a drug, 20,21 which is an ideal solution for obtaining a fine modulation over circadian time of biological clocks and has been employed in other oscillating systems. 22…”

Section: Introductionmentioning

confidence: 99%

Controlling the Circadian Clock with High Temporal Resolution through Photodosing

et al. 2019

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Circadian clocks, biological timekeepers that are present in almost every cell of our body, are complex systems whose disruption is connected to various diseases. Controlling cellular clock function with high temporal resolution in an inducible manner would yield an innovative approach for the circadian rhythm regulation. In the present study, we present structure-guided incorporation of photoremovable protecting groups into a circadian clock modifier, longdaysin, which inhibits casein kinase I (CKI). Using photodeprotection by UV or visible light (400 nm) as the external stimulus, we have achieved quantitative and light-inducible control over the CKI activity accompanied by an accurate regulation of circadian period in cultured human cells and mouse tissues, as well as in living zebrafish. This research paves the way for the application of photodosing in achieving precise temporal control over the biological timing and opens the door for chronophotopharmacology to deeper understand the circadian clock system.

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Optical Control of a Biological Reaction–Diffusion System

Cited by 28 publications

References 25 publications

Synthetic cell division via membrane-transforming molecular assemblies

Synthetic cell division via membrane-transforming molecular assemblies

<em>In Vitro</em> Reconstitution of Self-Organizing Protein Patterns on Supported Lipid Bilayers

Controlling the Circadian Clock with High Temporal Resolution through Photodosing

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