Protein Synthesis via <i>in Vitro</i> Transcription and Translation System inside Monodisperse Vesicles Fabricated by Microfluidics

In this study, we investigated the conditions for protein synthesis by an in vitro transcription and translation (IVTT) system within giant unilamellar vesicles (GUVs) produced with a microfluidic channel. The commercial IVTT system comprised of purified components (PURE system) and DNA encoding target protein was encapsulated in GUVs and incubated to synthesize the proteins. Synthesis of green fluorescent protein (GFP) and nanopore-forming α-Hemolysin were tested as the water-soluble protein and membrane protein, respectively. The stability of the GUVs and the efficiency of protein synthesis were assessed, focusing on variations in the concentration of the PURE system and the size of the GUVs. Our findings contribute to the development of homogeneous bioreactors and biosensors based on GUV technology.