Multiphase Water-in-Oil Emulsion Droplets for Cell-Free Transcription–Translation | Zendy

Paola Della Torre | Zendy; Christine D. Keating | Zendy; Sheref S. Mansy | Zendy

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Multiphase Water-in-Oil Emulsion Droplets for Cell-Free Transcription–Translation

Author(s) -

Paola Della Torre,

Christine D. Keating,

Sheref S. Mansy

Publication year - 2014

Publication title -

langmuir

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.042

H-Index - 333

eISSN - 1520-5827

pISSN - 0743-7463

DOI - 10.1021/la404146g

Subject(s) - emulsion , aqueous solution , dextran , ethylene glycol , aqueous two phase system , chemistry , phase (matter) , organelle , biophysics , chemical engineering , chromatography , biochemistry , organic chemistry , biology , engineering

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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