Light-Patterned Current Generation in a Droplet Bilayer Array

Schild, Vanessa Restrepo; Booth, Michael J.; Box, Stuart J.; Olof, Sam N.; Mahendran, Kozhinjampara R.; Bayley, Hagan

doi:10.1038/srep46585

Cited by 25 publications

(17 citation statements)

References 21 publications

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“…This high-resolution remote control of droplet networks is an important step forward in their development as biodevices. We have previously incorporated light-activated protein pumps into droplet arrays to generate patterned current generation 31 . In accord with our use of the model light-activated protein rsTagRFP, we might now incorporate these light-activated protein pumps into synthetic tissues to generate power at single droplet resolution.…”

Section: Discussionmentioning

confidence: 99%

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Light-patterning of synthetic tissues with single droplet resolution

Booth

Schild

Box

et al. 2017

Sci Rep

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Synthetic tissues can be generated by forming networks of aqueous droplets in lipid-containing oil. Each droplet contains a cell-free expression system and is connected to its neighbor through a lipid bilayer. In the present work, we have demonstrated precise external control of such networks by activating protein expression within single droplets, by using light-activated DNA to encode either a fluorescent or a pore-forming protein. By controlling the extent of activation, synthetic tissues were generated with graded levels of protein expression in patterns of single droplets. Further, we have demonstrated reversible activation within individual compartments in synthetic tissues by turning a fluorescent protein on-and-off. This is the first example of the high-resolution patterning of droplet networks, following their formation. Single-droplet control will be essential to power subsets of compartments within synthetic tissues or to stimulate subsets of cells when synthetic tissues are interfaced with living tissues.

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Section: Discussionmentioning

confidence: 99%

“…Data were acquired at 20 kHz with a 5 kHz filter through the ‘TecellaLab v0.90 type 2’ software. The data was in Tecella’s tlc binary file type and analysed and displayed by using custom software written in LabVIEW 31 .…”

Section: Methodsmentioning

confidence: 99%

Light-patterning of synthetic tissues with single droplet resolution

Booth

Schild

Box

et al. 2017

Sci Rep

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“…Considerable progress has been made in this area with engineered living cells [30,31], providing useful lessons for signaling by synthetic systems. A wide variety of receptors are available, from proteins such as bacteriorhodopsin, which can act as a light receptor [32], to completely synthetic receptors, which can respond to various inputs [33,34]. The fast transmission of signals through synthetic tissues can be electrical, as described earlier, while promising alternatives, including mechanical transmission [35] and the propagation of chemical waves [36], remain to be full exploited.…”

Section: Signaling In Synthetic Tissuesmentioning

confidence: 99%

“…In other cases, close integration might be required, which might be achieved, for example, by electrical coupling between the outermost bilayers of the two components [8]. Patterned electrical signals can be produced by droplet arrays [32] and printed versions might be used to control excitable tissues. The effects of weak electric fields on tissues and organs are an intriguing area of investigation that impacts development and tissue regeneration [42].…”

Section: Hybrid Tissuesmentioning

confidence: 99%

Synthetic tissues

Bayley

Hoskin

2019

Emerging Topics in Life Sciences

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While significant advances have been achieved with non-living synthetic cells built from the bottom-up, less progress has been made with the fabrication of synthetic tissues built from such cells. Synthetic tissues comprise patterned three-dimensional (3D) collections of communicating compartments. They can include both biological and synthetic parts and may incorporate features that do more than merely mimic nature. 3D-printed materials based on droplet-interface bilayers are the basis of the most advanced synthetic tissues and are being developed for several applications, including the controlled release of therapeutic agents and the repair of damaged organs. Current goals include the ability to manipulate synthetic tissues by remote signaling and the formation of hybrid structures with fabricated or natural living tissues.

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“…Cell-free protein synthesis of membrane proteins in a DIB has been successfully demonstrated using aHL, 15,29,41 Kcv, 15 LacY, 44 KcsA, and the hERG pore domain. 39 Usually an IVTT reaction is initiated in bulk and the mixture subsequently used to form DIBs.…”

Section: Insertion From Ivttmentioning

confidence: 99%

Probing membrane protein properties using droplet interface bilayers

Allen-Benton

Findlay

Booth

2019

Exp Biol Med (Maywood)

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Integral membrane proteins comprise a large proportion of drug targets, yet are challenging to study in vitro due to their amphiphilic nature. Conducting useful functional in vitro studies requires an artificial membrane that can mimic the lipid environment of the biogenic membrane. Droplet interface bilayer technology provides a method to form artificial bilayers with a robustness and physicochemical complexity that has not previously been possible, facilitating more sophisticated in vitro studies of membrane proteins. This mini-review examines functional studies of membrane proteins that utilize droplet interface bilayers to date and comments on possible directions of future research. Observations from our own laboratory regarding the study of a flippase protein in droplet interface bilayers are also presented. Impact statement The paper presents a comprehensive review of integral membrane protein studies utilizing droplet interface bilayers. Droplet interface bilayers are a novel method of constructing artificial lipid bilayers with enhanced stability and physicochemical complexity compared to existing methods. Their unique morphology also suggests applications in the construction of synthetic biological systems and protocells. As well as serving as a guide to in vitro membrane protein functional studies using droplet interface bilayers in the literature to date, a novel in vitro study of a flippase protein in a droplet interface bilayer is presented.

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Light-Patterned Current Generation in a Droplet Bilayer Array

Cited by 25 publications

References 21 publications

Light-patterning of synthetic tissues with single droplet resolution

Light-patterning of synthetic tissues with single droplet resolution

Synthetic tissues

Probing membrane protein properties using droplet interface bilayers

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