Protein synthesis with liposome fusion and fission by using the freeze-thaw method

For the experimental construction of artificial cell, it is a challenge to simultaneously supply the nutrients and lipids required for protein synthesis, gene replication, membrane growth, and fission. Inner reactions of liposomes are not permanent because of nutrient exhaustion since liposomes do not have pores or channels on their membrane for acquisition of nutrients. In this study, we demonstrated that the liposome containing in vitro translation system was fused with liposome encapsulating RNA by freeze and thaw for GFP synthesis. The fusion mixed lipid molecules on the two kinds of liposomes, followed by the fission. Consequently, we observed GFP synthesis inside the liposomes after liposome fusion and fission. This freeze and thaw method can be repeated, for the sustainable supplement of nutrients with liposome growth. We hope this method would achieve the ultimate goal of establishing the artificial cells that can acquire nutrients sustainably and proliferate by coupling protein synthesis and gene replication compatible with membrane growth.