ArticlePloS one2017
Protein aggregation with poly(vinyl) alcohol surfactant reduces double emulsion-encapsulated mammalian cell-free expression.
Article in PloS one, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Interfacial Interactions and Structural Evolution of Gelatin/Zein Nanofiber Composites Modulated by Poly(Vinyl Alcohol).Foods (Basel, Switzerland) · 2026Article
- Systematic investigation of double emulsion dewetting dynamics for the robust production of giant unilamellar vesicles.Lab on a chip · 2026Article
- Emerging Advances in Microfluidic Hydrogel Droplets for Tissue Engineering and STEM Cell Mechanobiology.Gels (Basel, Switzerland) · 2023Review
- Article
- Surfactant-free production of biomimetic giant unilamellar vesicles using PDMS-based microfluidics.Communications chemistry · 2021Article
- Bottom-Up Construction of Complex Biomolecular Systems With Cell-Free Synthetic Biology.Frontiers in bioengineering and biotechnology · 2020Review
- Reconstitution of Cell-cycle Oscillations in Microemulsions of Cell-free Xenopus Egg Extracts.Journal of visualized experiments : JoVE · 2018Article
- Microfluidic fabrication of microparticles for biomedical applications.Chemical Society reviews · 2018Review
- Article
- Systems and synthetic biology approaches in understanding biological oscillators.Quantitative biology (Beijing, China) · 2018Article
- Article
- Simultaneous monitoring of transcription and translation in mammalian cell-free expression in bulk and in cell-sized droplets.Synthetic biology (Oxford, England) · 2018Article
- Extrinsic stochastic factors (solute partition) in gene expression inside lipid vesicles and lipid-stabilized water-in-oil droplets: a review.Synthetic biology (Oxford, England) · 2018Review
- Bottom-up synthetic biology: modular design for making artificial platelets.Physical biology · 2017Article
- Cell-sized mechanosensitive and biosensing compartment programmed with DNA.Chemical communications (Cambridge, England) · 2017Article
Corrections and comments
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Authors and funding
5 authors.
Funding
Abstract
Development of artificial cell models requires encapsulation of biomolecules within membrane-bound compartments. There have been limited studies of using mammalian cell-free expression (CFE) system as the 'cytosol' of artificial cells. We exploit glass capillary droplet microfluidics for the encapsulation of mammalian CFE within double emulsion templated vesicles. The complexity of the physicochemical properties of HeLa cell-free lysate poses a challenge compared with encapsulating simple buffer solutions. In particular, we discovered the formation of aggregates in double emulsion templated vesicles encapsulating mammalian HeLa CFE, but not with bacterial CFE. The aggregates did not arise from insolubility of the proteins made from CFE nor due to the interaction of mammalian CFE with the organic solvents in the middle phase of the double emulsions. We found that aggregation is dependent on the concentration of poly(vinyl) alcohol (PVA) surfactant, a critical double emulsion-stabilizing surfactant, and the lysate concentration in mammalian CFE. Despite vesicle instability and reduced protein expression, we demonstrate protein expression by encapsulating mammalian CFE system. Using mass spectrometry and Western blot, we identified and verified that actin is one of the proteins inside the mammalian CFE that aggregated with PVA surfactant. Our work establishes a baseline description of mammalian CFE system encapsulated in double emulsion templated vesicles as a platform for building artificial cells.
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Registered trials
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