Toward an artificial cell based on gene expression in vesicles
- 1 September 2005
- journal article
- Published by IOP Publishing in Physical Biology
- Vol. 2 (3), P1-P8
- https://doi.org/10.1088/1478-3975/2/3/p01
Abstract
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.Keywords
This publication has 24 references indexed in Scilit:
- A vesicle bioreactor as a step toward an artificial cell assemblyProceedings of the National Academy of Sciences, 2004
- Principles of cell-free genetic circuit assemblyProceedings of the National Academy of Sciences, 2003
- Engineering asymmetric vesiclesProceedings of the National Academy of Sciences, 2003
- Artificial cells: prospects for biotechnologyTrends in Biotechnology, 2002
- Cell-free translation reconstituted with purified componentsNature Biotechnology, 2001
- Chemical Transformations in Individual Ultrasmall Biomimetic ContainersScience, 1999
- Man-made cell-like compartments for molecular evolutionNature Biotechnology, 1998
- Effect of the ATP level on the overall protein biosynthesis rate in a wheat germ cell-free systemBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1996
- A Continuous Cell-Free Translation System Capable of Producing Polypeptides in High YieldScience, 1988
- Energy charge of the adenylate pool as a regulatory parameter. Interaction with feedback modifiersBiochemistry, 1968