Cytoskeletal Protein Expression and its Association within the Hydrophobic Membrane of Artificial Cell Models

Martino, Chiara; Horsfall, Louise; Chen, Yan; Chanasakulniyom, Mayuree; Paterson, David J.; Brunet, Adrien; Rosser, Susan J.; Yang, Yuan; Cooper, Jonathan M.

doi:10.1002/cbic.201200038

Cited by 13 publications

(14 citation statements)

References 26 publications

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“…Lipid-free experiments performed with water/oil/water (w/o/w) double emulsions (Martino et al 2012a), and water/oil (w/o) droplets (Chanasakulniyom et al 2012), showed that a freshly synthesised fusion protein, comprised of MreB joined to red fluorescent protein (RFP), accumulated at the surfactant-based emulsion interface.…”

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confidence: 99%

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Synthetic Plants

2016

Synthetic Biology Handbook

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Section: Downloaded By [University College London] At 07:47 24 June mentioning

confidence: 99%

“…When MreB-RFP protein was synthesised inside microfluidic-produced polymersomes, its distribution was partially on the membrane, partially localised in the polymersome core (Martino et al 2012a).…”

Section: Downloaded By [University College London] At 07:47 24 June mentioning

confidence: 99%

Synthetic Plants

2016

Synthetic Biology Handbook

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“…[5][6][7][8] Methods involved with combining microdroplet technologies with synthetic biology techniques have proved particularly challenging and innovative. [9][10][11][12][13][14][15][16][17] Customised DNA templates and commercially available cell-free expression systems have been combined and encapsulated in microdroplets for the synthesis of enzyme and water soluble proteins, such as green fluorescent protein (GFP) and red shifted GFP (rsGFP) . [13][14][15][16][17] Developments in synthetic biology have potential applications in the construction of artificial cells and more broadly in the production of synthetically created biological structures in artificial environments, like single-emulsion droplets.…”

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confidence: 99%

Expression of membrane-associated proteins within single emulsion cell facsimiles

Chanasakulniyom

Martino

Paterson

et al. 2012

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MreB is a structural membrane-associated protein which is one of the key components of the bacterial cytoskeleton. Although it plays an important role in shape maintenance of rod-like bacteria, the understanding of its mechanism of action is still not fully understood. This study shows how segmented flow and microdroplet technology can be used as a new tool for biological in vitro investigation of this protein. In this paper, we demonstrate cell-free expression in a single emulsion system to express red fluorescence protein (RFP) and MreB linked RFP (MreB-RFP). We follow the aggregation and localisation of the fusion protein MreB-RFP in this artificial cell-like environment. The expression of MreB-RFP in single emulsion droplets leads to the formation of micrometer-scale protein patches distributed at the water/oil interface.

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“…[7][8][9] These studies establish a solid foundation for the engineering of more complex cellular nanosystems. In fact, it is now possible to combine individual biomimetic components to create multifunctional cellular systems that mimic functions of natural cells, including gene expression, [10][11][12][13][14] membrane transport, 14 and subcellular localization. 15 Specifically, scientists have synthesized artificial cells (or protocells) by integrating synthetic gene circuits and bio-membranes.…”

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The engineering of artificial cellular nanosystems using synthetic biology approaches

Tan

2014

WIREs Nanomed Nanobiotechnol

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Artificial cellular systems are minimal systems that mimic certain properties of natural cells, including signaling pathways, membranes, and metabolic pathways. These artificial cells (or protocells) can be constructed following a synthetic biology approach by assembling biomembranes, synthetic gene circuits, and cell-free expression systems. As artificial cells are built from bottom-up using minimal and a defined number of components, they are more amenable to predictive mathematical modeling and engineered controls when compared with natural cells. Indeed, artificial cells have been implemented as drug delivery machineries and in situ protein expression systems. Furthermore, artificial cells have been used as biomimetic systems to unveil new insights into functions of natural cells, which are otherwise difficult to investigate owing to their inherent complexity. It is our vision that the development of artificial cells would bring forth parallel advancements in synthetic biology, cell-free systems, and in vitro systems biology. For further resources related to this article, please visit the WIREs website. Conflict of interests: The authors declare that they have no competing financial interests.

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Cytoskeletal Protein Expression and its Association within the Hydrophobic Membrane of Artificial Cell Models

Cited by 13 publications

References 26 publications

Synthetic Plants

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Expression of membrane-associated proteins within single emulsion cell facsimiles

The engineering of artificial cellular nanosystems using synthetic biology approaches

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