To expand the range of practical applications of artificial
cells,
it is important to standardize the production process of giant (cell-sized)
vesicles that encapsulate reconstituted biochemical reaction systems.
For this purpose, a rapidly developing microfluidics-based giant vesicle
generation system is a promising approach, similar to the droplet
assay systems that are already widespread in the market. In this study,
we examined the composition of the solutions used to generate vesicles
encapsulating the in vitro transcription–translation
(IVTT) system. We show that tuning of the lipid composition and adding
poly(vinyl alcohol) to the outer solution improved the stability of
the transition process into the lipid membrane so that protein synthesis
proceeded in vesicles. The direct integration of α-hemolysin
nanopores synthesized in situ was also demonstrated.
These protein-synthesizing monodisperse giant vesicles can be prepared
by using a simple microfluidic fabrication/operation with a commercial
IVTT system.