RT Journal Article
SR Electronic
T1 Genetically controlled membrane synthesis in liposomes
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.03.29.013300
DO 10.1101/2020.03.29.013300
A1 Duco Blanken
A1 David Foschepoth
A1 Adriana Calaça Serrão
A1 Christophe Danelon
YR 2020
UL http://biorxiv.org/content/early/2020/03/30/2020.03.29.013300.abstract
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.