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Systems and Synthetic Biology

, Volume 4, Issue 3, pp 149–156 | Cite as

Synthetic biology of minimal living cells: primitive cell models and semi-synthetic cells

  • Pasquale Stano
Research Article

Abstract

This article summarizes a contribution presented at the ESF 2009 Synthetic Biology focused on the concept of the minimal requirement for life and on the issue of constructive (synthetic) approaches in biological research. The attempts to define minimal life within the framework of autopoietic theory are firstly described, and a short report on the development of autopoietic chemical systems based on fatty acid vesicles, which are relevant as primitive cell models is given. These studies can be used as a starting point for the construction of more complex systems, firstly being inspired by possible origins of life scenarioes (and therefore by considering primitive functions), then by considering an approach based on modern biomacromolecular-encoded functions. At this aim, semi-synthetic minimal cells are defined as those man-made vesicle-based systems that are composed of the minimal number of genes, proteins, biomolecules and which can be defined as living. Recent achievements on minimal sized semi-synthetic cells are then discussed, and the kind of information obtained is recognized as being distinctively derived by a constructive approach. Synthetic biology is therefore a fundamental tool for gaining basic knowledge about biosystems, and it should not be confined at all to the engineering side.

Keywords

Autopoiesis Semi-synthetic minimal cells Cell-free protein synthesis Self-reproduction Minimal cell size 

Notes

Acknowledgments

This work has been presented at the Second European Conference on Synthetic Biology (ESF ECSB II: Design, Programming and Optimization of Biological Systems, Sant Feliux de Guixol, Spain, 29 March-03 April, 2009); it collects the recent results of Luisi’s group at ETH Zurich and University of RomaTre. I greatly thank Pier Luigi Luisi for inspiring this research and for stimulating discussions. Tereza Souza (University of Jena) and Yutetsu Kuruma (University of Tokyo) are also acknowledged as co-authors of the original papers on minimal cell size and lipid-synthesizing minimal cell. This work has been funded by the SYNTHCELLS project (Approaches to the Bioengineering of Synthetic Minimal Cells, EU Grant #FP6043359); by the Human Frontiers Science Program (RGP0033/2007-C) and by the Italian Space Agency (Grant Nr. I/015/07/0). It is also developed within the COST Systems Chemistry CM0703 Action.

References

  1. Amidi M, de Raad M, de Graauw H, van Ditmarsch D, Hennink WE, Crommelin DJA, Mastrobattista E (2009) Optimization and quantification of protein synthesis inside liposomes. J Lipos Res (in press)Google Scholar
  2. Bachmann PA, Luisi PL, Lang J (1992) Autocatalytic self-replicating micelles as models for prebiotic structures. Nature 357:57–59CrossRefGoogle Scholar
  3. Berclaz N, Muller M, Walde P, Luisi PL (2001) Growth and transformation of vesicles studied by ferritin labeling and cryotransmission electron microscopy. J Phys Chem B 105:1056–1064CrossRefGoogle Scholar
  4. Bitbol M, Luisi PL (2004) Autopoiesis with or without cognition: defining life at its edge. J Royal Soc Interface 1:99–107CrossRefGoogle Scholar
  5. Blochliger E, Blocher M, Walde P, Luisi PL (1998) Matrix effect in the size distribution of fatty acid vesicles. J Phys Chem 102:10383–10390Google Scholar
  6. Chen IA, Szostak JW (2004) A kinetic study of the growth of fatty acid vesicles. Biophys J 87:988–998CrossRefPubMedGoogle Scholar
  7. Damiano L (2009) Unità in dialogo. Un nuovo stile per la conoscenza. Mondadori, MilanoGoogle Scholar
  8. De Lorenzo V, Danchin A (2008) Synthetic biology: discovering new worlds and new words. EMBO Rep 9:9Google Scholar
  9. Deamer DW, Dworkin JP (2005) Chemistry and physics of primitive membranes. Top Curr Chem 259:1–27CrossRefGoogle Scholar
  10. Foster AC, Church GM (2006) Towards synthesis of a minimal cell. Mol Syst Biol 2. doi: 10.1038/msb4100090
  11. Gil R, Silva FJ, Peretó J, Moya A (2004) Determination of the core of a minimal bacteria gene set. Microbiol Mol Biol Rev 68:518–537CrossRefPubMedGoogle Scholar
  12. Hasoda K, Sunami T, Kazuta Y, Matsuura T, Suzuki H, Yomo T (2008) Quantitative study of the structure of multilamellar giant liposomes as a container of protein synthesis reaction. Langmuir 24:13540–13548CrossRefGoogle Scholar
  13. Ishikawa K, Sato K, Shima Y, Urabe I, Yomo T (2004) Expression of a cascading genetic network within liposomes. FEBS Lett 576:387–390CrossRefPubMedGoogle Scholar
  14. Kajander EO, Ciftcioglu N (1998) Nanobacteria: an alternative mechanism for pathogenic intra- and extracellular calcification and stone formation. Proc Natl Acad Sci USA 95:8274–8279CrossRefPubMedGoogle Scholar
  15. Kita H, Matsuura T, Sunami T, Hosoda K, Ichihashi N, Tsukada K, Urabe I, Yomo T (2008) Replication of genetic information with self-encoded replicase in liposomes. Chembiochem 9:2403–2410CrossRefPubMedGoogle Scholar
  16. Knoll A, Osborn MJ, Baross J, Berg HC, Pace NR, Sogin M (eds) (1999) Size limits of very small microorganisms. National Academic Press, WashingtonGoogle Scholar
  17. Kuruma Y, Stano P, Ueda T, Luisi PL (2009) A synthetic biology approach to the construction of membrane proteins in semi-synthetic minimal cells. Biochim Biophys Acta 1788:567–574CrossRefPubMedGoogle Scholar
  18. Liu AP, Fletcher DA (2009) Biology under construction: in vitro reconstruction of cellular function. Nature Rev 10:644–650CrossRefGoogle Scholar
  19. Lonchin S, Luisi PL, Walde P, Robinson BH (1999) A matrix effect in mixed phospholipid/fatty acid vesicle formation. J Phys Chem B 103:10910–10916CrossRefGoogle Scholar
  20. Luisi PL (2007) Chemical aspects of synthetic biology. Chem Biodiv 4:603–621CrossRefGoogle Scholar
  21. Luisi PL, Varela FJ (1990) Self-replicating micelles–a chemical version of minimal autopoietic systems. Orig Life Evol Biosph 19:633–643CrossRefGoogle Scholar
  22. Luisi PL, Walde P, Oberholzer T (1999) Lipid vesicles as possible intermediates in the origin of life. Curr Opin Coll Inter Sci 4:33–39CrossRefGoogle Scholar
  23. Luisi PL, Oberholzer T, Lazcano A (2002) The notion of a DNA minimal cell: a general discourse and some guidelines for an experimental approach. Helv Chim Acta 85:1759–1777CrossRefGoogle Scholar
  24. Luisi PL, Ferri F, Stano P (2006) Approaches to semi-synthetic minimal cells: a review. Naturwissenschaften 93:1–13CrossRefPubMedGoogle Scholar
  25. Luisi PL, Souza T, Stano P (2008) Vesicle behavior: in search of explanations. J Phys Chem B 112:14655–14664CrossRefPubMedGoogle Scholar
  26. Mansy SS, Szostak JW (2009) Reconstructing the emergence of cellular life through the synthesis of model protocells. Cold Spring Harbor Symposia on Quantitative Biology, vol. LXXIV, Cold Spring Harbor Laboratory Press, pp 1–9Google Scholar
  27. Maturana HR, Varela FJ (1980) Autopoiesis and cognition: the realization of the living. Reidel, DordrechtGoogle Scholar
  28. Moore PB (1999) A biophysical chemist’s thoughts on cell size. In: Knoll A, Osborn MJ, Baross J, Berg HC, Pace NR, Sogin M (eds) Size limits of very small microorganisms. National Academic Press, WashingtonGoogle Scholar
  29. Morowitz H (1992) Beginnings of cellular life. Metabolism recapitulates biogenesis. Yale University Press, New HavenGoogle Scholar
  30. Murtas G, Kuruma Y, Bianchini P, Diaspro A, Luisi PL (2007) Protein synthesis in liposomes with a minimal set of enzymes. Biochem Biophys Res Comm 363:12–17CrossRefPubMedGoogle Scholar
  31. Noireaux V, Libchaber A (2004) A vesicle bioreactor as a step toward an artificial cell assembly. Proc Natl Acad Sci USA 101:17669–17674CrossRefPubMedGoogle Scholar
  32. Nomura SM, Tsumoto K, Hamada T, Akiyoshi K, Nakatani Y, Yoshikawa K (2003) Gene expression within cell-sized lipid vesicles. ChemBioChem 4:1172–1175CrossRefPubMedGoogle Scholar
  33. Oberholzer T, Luisi PL (2002) The use of liposomes for constructing cell models. J Biol Phys 28:733–744CrossRefGoogle Scholar
  34. Oberholzer T, Wick R, Luisi PL, Biebricher CK (1995) Enzymatic RNA replication in self- reproducing vesicles: an approach to a minimal cell. Biochem Biophys Res Comm 207:250–257CrossRefPubMedGoogle Scholar
  35. Oberholzer T, Nierhaus KH, Luisi PL (1999) Protein expression in liposomes. Biochem Biophys Res Commun 261:238–241CrossRefPubMedGoogle Scholar
  36. Pautot S, Frisken BJ, Weitz DA (2003) Production of unilamellar vesicles using an inverted emulsion. Langmuir 19:2870–2879CrossRefGoogle Scholar
  37. Pohorille A, Deamer D (2002) Artificial cells: prospects for biotechnology. Trends Biotechnol 20:123–128CrossRefPubMedGoogle Scholar
  38. Rasi S, Mavelli F, Luisi PL (2003) Cooperative micelle binding and matrix effect in oleate vesicle formation. J Phys Chem B 107:14068–14076CrossRefGoogle Scholar
  39. Rogerson ML, Robinson BH, Bucak S, Walde P (2006) Kinetic studies of the interaction of fatty acids with phosphatidylcholine vesicles (liposomes). Colloids Surf B Biointerfaces 48:24–34CrossRefPubMedGoogle Scholar
  40. Saito H, Yamada A, Ohmori R, Kato Y, Yamanaka T, Yoshikawa K, Inoue T (2007) Towards constructing synthetic cells: RNA/RNP evolution and cell-free translational systems in giant liposomes. In: International symposium on micro-nanomechatronics and human science, MHS 07, pp 286–291Google Scholar
  41. Saito H, Kato Y, Le Berre M, Yamada A, Inoue T, Yoshikawa K, Baigl D (2009) Time-resolved tracking of a minimum gene expression system reconstituted in giant liposomes. ChemBioChem 10:1640–1643CrossRefPubMedGoogle Scholar
  42. Schwille P, Diez S (2009) Synthetic biology of minimal systems. Crit Rev Biochem Mol Biol 44:223–242CrossRefPubMedGoogle Scholar
  43. Shapiro R (2007) A simpler origin for life. Sci Am 296:46–53CrossRefPubMedGoogle Scholar
  44. Shimizu Y, Inoue A, Tomari Y, Suzuki T, Yokogawa T, Nishikawa K (2001) Ueda T: cell-free translation reconstituted with purified components. Nat Biotechnol 19:751–755CrossRefPubMedGoogle Scholar
  45. Souza T, Stano P, Luisi PL (2009) The minimal size of liposome-based model cells brings about a remarkably enhanced entrapment and protein synthesis. ChemBioChem 10:1056–1063CrossRefGoogle Scholar
  46. Stano P (2009) Self-reproduction of vesicles and other compartments: a review. In: Birdi KS (ed) CRC handbook of surface and colloid chemistry, 3rd edn. CRC PRESS, Taylor and Francis Group LLC, Boca Raton, pp 681–701Google Scholar
  47. Stano P, Luisi PL (2008) Self-reproduction of micelles, reverse micelles and vesicles. Compartments disclose a general transformation pattern. In: Leitmannova Liu A (ed) Advances on planar lipid bilayers and liposomes. Elsevier, Amsterdam, pp 221–263Google Scholar
  48. Stano P, Wehrli E, Luisi PL (2006) Insights on the oleate vesicles self-reproduction. J Phys Condens Matter 18:S2231–S2238CrossRefGoogle Scholar
  49. Szostak JW, Bartel DP, Luisi PL (2001) Synthesizing life. Nature 409:387–390CrossRefPubMedGoogle Scholar
  50. Walde P (2006) Surfactant assemblies and their various possible roles for the origin(s) of life. Orig Life Evol Biosph 36:109–150CrossRefPubMedGoogle Scholar
  51. Walde P, Goto A, Monnard PA, Wessicken M, Luisi PL (1994a) Oparin’s reactions revisited: enzymatic synthesis of poly(adenylic acid) in micelles and self-reproducing vesicles. J Am Chem Soc 116:7541–7544CrossRefGoogle Scholar
  52. Walde P, Wick R, Fresta M, Mangone A, Luisi PL (1994b) Autopoietic self-reproduction of fatty acid vesicles. J Am Chem Soc 116:11649–11654CrossRefGoogle Scholar
  53. Yu W, Sato K, Wakabayashi M, Nakatshi T, Ko-Mitamura EP, Shima Y, Urabe I, Yomo T (2001) Synthesis of functional protein in liposome. J Biosci Bioeng 92:590–593CrossRefPubMedGoogle Scholar
  54. Zhang Y, Ruder WC, LeDuc PR (2008) Artificial cells: building bioinspired systems using small-scale biology. TRENDS Biotechnol 26:14–20CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  1. 1.Biology DepartmentUniversity of RomaTreRomeItaly

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