Multiphase water-in-oil emulsion droplets for cell-free transcription-translation

Paola Torre; Christine D. Keating; Sheref S. Mansy

doi:10.1021/la404146g

Multiphase water-in-oil emulsion droplets for cell-free transcription-translation

Paola Torre, Christine D. Keating, Sheref S. Mansy

Research output: Contribution to journal › Article › peer-review

80 Scopus citations

Abstract

The construction of genetically encoded cellular mimics in compartments containing organized synthetic cytosols is desirable for the development of artificial cells. Phase separated aqueous domains were placed within water-in-oil emulsion droplets in a manner compatible with transcription and translation machinery. Aqueous two-phase and three-phase systems (ATPS and A3PS) were assembled with dextran, poly(ethylene glycol), and Ficoll. Aqueous two-phase systems were capable of supporting the cell-free expression of protein within water droplets, whereas the aqueous three-phase-based system did not give rise to detectable protein synthesis. The expressed protein preferentially partitioned to the dextran-enriched phase. The system could serve as a foundation for building cellular mimics with liquid organelles.

Original language	English (US)
Pages (from-to)	5695-5699
Number of pages	5
Journal	Langmuir
Volume	30
Issue number	20
DOIs	https://doi.org/10.1021/la404146g
State	Published - May 27 2014

All Science Journal Classification (ASJC) codes

General Materials Science
Condensed Matter Physics
Surfaces and Interfaces
Spectroscopy
Electrochemistry

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10.1021/la404146g

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AB - The construction of genetically encoded cellular mimics in compartments containing organized synthetic cytosols is desirable for the development of artificial cells. Phase separated aqueous domains were placed within water-in-oil emulsion droplets in a manner compatible with transcription and translation machinery. Aqueous two-phase and three-phase systems (ATPS and A3PS) were assembled with dextran, poly(ethylene glycol), and Ficoll. Aqueous two-phase systems were capable of supporting the cell-free expression of protein within water droplets, whereas the aqueous three-phase-based system did not give rise to detectable protein synthesis. The expressed protein preferentially partitioned to the dextran-enriched phase. The system could serve as a foundation for building cellular mimics with liquid organelles.

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Multiphase water-in-oil emulsion droplets for cell-free transcription-translation

Abstract

All Science Journal Classification (ASJC) codes

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