Identification of conditions for efficient cell-sized liposome preparation using commercially available reconstituted in vitro transcription-translation system

Available online 14 November 2021 Attempts to create complex molecular systems that mimic parts of cellular systems using a bottom-up approach have become important in the field of biology. Among various molecular systems, in vitro protein synthesis inside lipid vesicles (liposomes), which we refer to as the artificial cell, has become an attractive system because it possesses two fundamental features of living cells: central dogma, and compartmentalization. Here, we investigated the effect of altering the amount or concentration of four constituents of the artificial cell consisting of a commercially available reconstituted in vitro transcription-translation (IVTT) system. As this IVTT system is available worldwide, the results will be useful to the scientific community when shared, unlike those from a lab-made IVTT system. We succeeded in revealing the effect and trend of altering each parameter and identified a suitable condition for preparing liposomes that are unilamellar and can synthesize proteins equally as well as the original IVTT system. Because the commercially available reconstituted IVTT system is an important standardization tool and the constituents can be adjusted as desired, our results will be useful for the bottom-up creation of more complex molecular systems. (c) 2021, The Society for Biotechnology, Japan. All rights reserved.

Automated summary

Efforts to create complex molecular systems from the bottom-up in biology have gained importance. One such system is the artificial cell, where protein synthesis occurs inside lipid vesicles. By altering the amount or concentration of specific constituents in the artificial cell, researchers have identified suitable conditions for preparing unilamellar liposomes capable of synthesizing proteins as well as the original system. These results are valuable for the bottom-up creation of more complex molecular systems.