Toward an artificial cell based on gene expression in vesicles.
We present a new experimental approach to build an artificial cell using the translation machinery of a cell-free expression system as the hardware and a DNA synthetic genome as the software. This approach, inspired by the self-replicating automata of von Neumann, uses cytoplasmic extracts, encapsulated in phospholipid vesicles, to assemble custom-made genetic circuits to develop the functions of a minimal cell. Although this approach can find applications, especially in biotechnology, the primary goal is to understand how a DNA algorithm can be designed to build an operating system that has some of the properties of life. We provide insights on this cell-free approach as well as new results to transform step by step a long-lived vesicle bioreactor into an artificial cell. We show how the green fluorescent protein can be anchored to the membrane and we give indications of a possible insertion mechanism of integral membrane proteins. With vesicles composed of different phospholipids, the fusion protein alpha-hemolysin-eGFP can be expressed to reveal patterns on the membrane. The specific degradation complex ClpXP from E. coli is introduced to create a sink for the synthesized proteins. Perspectives and subsequent limitations of this approach are discussed.
Noireaux
Vincent
V
Center for Studies in Physics and Biology, The Rockefeller University, New York, NY 10021, USA. noireav@mail.rockefeller.edu
Bar-Ziv
Roy
R
Godefroy
Jeremy
J
Salman
Hanna
H
Libchaber
Albert
A
D002474:Cell-Free System; D002477:Cells / Q000737:chemistry* / Q000378:metabolism; D003593:Cytoplasm; D004247:DNA / Q000138:chemical synthesis; D015870:Gene Expression; D010743:Phospholipids; D014176:Protein Biosynthesis; D053835:Unilamellar Liposomes
D010743:Phospholipids; D053835:Unilamellar Liposomes; D004247:DNA