PT - JOURNAL ARTICLE AU - Duco Blanken AU - David Foschepoth AU - Adriana Calaça Serrão AU - Christophe Danelon TI - Genetically controlled membrane synthesis in liposomes AID - 10.1101/2020.03.29.013300 DP - 2020 Jan 01 TA - bioRxiv PG - 2020.03.29.013300 4099 - http://biorxiv.org/content/early/2020/03/30/2020.03.29.013300.short 4100 - http://biorxiv.org/content/early/2020/03/30/2020.03.29.013300.full AB - Lipid membrane, nucleic acids, proteins, and metabolism are essential ingredients for life. Synthetic cellular systems emulating the fundamental properties of living cells must therefore be built upon these functional elements. In this work, phospholipid-producing enzymes encoded in a synthetic minigenome are cell-free expressed within liposome compartments. The de novo synthesized metabolic pathway converts precursors into a variety of lipids, including the constituents of the parental liposome. Balanced production of phosphatidylethanolamine and phosphatidylglycerol is realized, owing to transcriptional regulation of specific genes activity combined with a metabolic feedback mechanism. Fluorescence-based methods are developed to image the synthesis and membrane incorporation of phosphatidylserine at the single liposome level. Our results provide experimental evidence for DNA-programmed membrane synthesis in a minimal cell model. Strategies are discussed to alleviate current limitations toward more effective liposome growth and self-reproduction.