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Microfluidic Tools for

Bottom-Up Synthetic

Cellularity

Stano, P. Is Research on “Synthetic Cells” Moving to the Next Level? Life 2019 , 9 , 3.

https://doi.org/10.3390/life

poly( N-

isopropylacrylamide) PEG-bis( N- succinimi dyl succinate)

S. Levine, B.D. Bowen, S.J. Partridge Colloids Surf., 38 (1989), pp. 325- Thompson, Kate L., Mark Williams, and Steven P. Armes. "Colloidosomes: synthesis, properties and

applications." Journal of colloid and interface

Microfluidics Meets

Artificial Cells

Scott, S.M.; Ali, Z. Fabrication Methods for Microfluidic Devices: An

Overview. Micromachi

nes 2021 , 12 , 319.

https://doi.org/10. /mi

Deshpande, S., Caspi, Y., Meijering,

A. et al. Octanol-

assisted liposome assembly on chip.

Nat Commun 7 ,

https://doi.org/10. 38/ncomms

PEG(5000)- b- PLA(5000) dissolved in a mixture of toluene and chloroform

FITC-

Dextran

• HPTS in water PVA aqueous solution Microfluidic Fabrication of Monodisperse Biocompatible and Biodegradable Polymersomes with Controlled Permeability, Ho Cheung Shum, et al., Journal of the American Chemical Society 2008130 (29), 9543-9549, DOI: 10.1021/ja802157y van Swaay, Dirk; Tang, T.-Y. Dora; Mann, Stephen; de Mello, Andrew (2015). Microfluidic Formation of Membrane-Free Aqueous Coacervate Droplets in Water. Angewandte Chemie International Edition, 54(29), 8398–8401. doi:10.1002/anie.

Incorporating Reactions

Into Droplets

Proteinosomes offer an alternative route to membrane activation, where chemically modified enzymes can be rendered amphiphilic by the addition of poly( N- isopropylacrylamide) (PNIPAAm) chains and then used to generate a stable water-in- oil emulsion. The self-assembled protein-conjugates can further be chemically cross-linked and transferred into water to generate micron-sized water-in-water compartments with an enzymatically active membrane Ugrinic, M., Zambrano, A., Berger, S., Mann, S., Tang, T.D., and Demello, A. (2018). Microfluidic formation of proteinosomes. Chem. Commun. 54, 287– Huang, X., Li, M., Green, D. et al. Interfacial assembly of protein–polymer nano- conjugates into stimulus-responsive biomimetic protocells. Nat Commun 4 ,

(a) Positive DEP occurs when the particle exhibits a larger polarisability than the surrounding medium. Positive DEP is directed towards the stronger electric field. (b) Negative DEP occurs if the particle is less polarisable than the

Higher-order

Structures