Naturwissenschaften

, Volume 93, Issue 1, pp 1–13

Approaches to semi-synthetic minimal cells: a review

  • Pier Luigi Luisi
  • Francesca Ferri
  • Pasquale Stano
Review

Abstract

Following is a synthetic review on the minimal living cell, defined as an artificial or a semi-artificial cell having the minimal and sufficient number of components to be considered alive. We describe concepts and experiments based on these constructions, and we point out that an operational definition of minimal cell does not define a single species, but rather a broad family of interrelated cell-like structures. The relevance of these researches, considering that the minimal cell should also correspond to the early simple cell in the origin of life and early evolution, is also explained. In addition, we present detailed data in relation to minimal genome, with observations cited by several authors who agree on setting the theoretical full-fledged minimal genome to a figure between 200 and 300 genes. However, further theoretical assumptions may significantly reduce this number (i.e. by eliminating ribosomal proteins and by limiting DNA and RNA polymerases to only a few, less specific molecular species). Generally, the experimental approach to minimal cells consists in utilizing liposomes as cell models and in filling them with genes/enzymes corresponding to minimal cellular functions. To date, a few research groups have successfully induced the expression of single proteins, such as the green fluorescence protein, inside liposomes. Here, different approaches are described and compared. Present constructs are still rather far from the minimal cell, and experimental as well as theoretical difficulties opposing further reduction of complexity are discussed. While most of these minimal cell constructions may represent relatively poor imitations of a modern full-fledged cell, further studies will begin precisely from these constructs. In conclusion, we give a brief outline of the next possible steps on the road map to the minimal cell.

References

  1. 1.
    Bachmann PA, Luisi PL, Lang J (1992) Autocatalytic self-replicating micelles as models for prebiotic structures. Nature 357:57–59CrossRefGoogle Scholar
  2. 2.
    Berclaz N, Mueller 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
  3. 3.
    Bloechliger 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
  4. 4.
    Calderone CT, Liu DR (2004) Nucleic-acid-templated synthesis as a model system for ancient translation. Curr Opin Chem Biol 8:645–653PubMedCrossRefGoogle Scholar
  5. 5.
    Cello J, Paul AV, Wimmer E (2002) Chemical synthesis of poliovirus cDNA: generation of infectious virus in the absence of natural template. Science 297:1016–1018PubMedCrossRefGoogle Scholar
  6. 6.
    Chakrabarti AC, Breaker RR, Joyce GF, Deamer DW (1994) Production of RNA by a polymerase protein encapsulated within phospholipid vesicles. J Mol Evol 39:555–559PubMedCrossRefGoogle Scholar
  7. 7.
    Dyson FJ (1982) A model for the origin of life. J Mol Evol 18:344–350PubMedCrossRefGoogle Scholar
  8. 8.
    Fischer A, Franco A, Oberholzer T (2002) Giant vesicles as microreactors for enzymatic mRNA synthesis. ChemBioChem 3:409–417PubMedCrossRefGoogle Scholar
  9. 9.
    Fraser CM, Gocayne JD, White O, Adams MD, Clayton RA, Fleischmann RD, Bult CJ, Kerlavage AR, Sutton G, Kelley JM et al (1995) The minimal gene complement of Mycoplasma genitalium. Science 270:397–403PubMedCrossRefGoogle Scholar
  10. 10.
    Frick DN, Richardson CC (2001) DNA primases. Annu Rev Biochem 70:39–80PubMedCrossRefGoogle Scholar
  11. 11.
    Gavrilova LP, Kostiashkina OE, Koteliansky VE, Rutkevitch NM, Spirin AS (1976) Factor-free (non-enzymic) and factor-dependent systems of translation of polyuridylic acid by Escherichia coli ribosomes. J Mol Biol 101:537–552PubMedCrossRefGoogle Scholar
  12. 12.
    Gil R, Sabater-Munoz B, Latorre A, Silva FJ, Moya A (2002) Extreme genome reduction in Buchnera spp: toward the minimal genome needed for symbiotic life. PNAS 99:4454–4458PubMedCrossRefGoogle Scholar
  13. 13.
    Gil R, Silva FJ, Peretó J, Moya A (2004) Determination of the core of a minimal bacteria gene set. Microbiol Mol Biol Rev 68:518–537PubMedCrossRefGoogle Scholar
  14. 14.
    Glade N, Demongeot J, Tabony J (2004) Microtubule self-organisation by reaction–diffusion processes causes collective transport and organisation of cellular particles. BMC Cell Biol 5:23. DOI 10.1186/1471-2121-5-23PubMedCrossRefGoogle Scholar
  15. 15.
    Hutchinson CA, Peterson SN, Gill SR, Cline RT, White O, Fraser CM, Smith HO, Venter JC (1999) Global transposon mutagenesis and a minimal Mycoplasma genome. Science 286:2165–2169CrossRefGoogle Scholar
  16. 16.
    Ishikawa K, Sato K, Shima Y, Urabe I, Yomo T (2004) Expression of a cascading genetic network within liposomes. FEBS Lett 576:387–390PubMedCrossRefGoogle Scholar
  17. 17.
    Islas S, Becerra A, Luisi PL, Lazcano A (2004) Comparative genomics and the gene complement of a minimal cell. Orig Life Evol Biosph 34:243–256PubMedCrossRefGoogle Scholar
  18. 18.
    Itaya M (1995) An estimation of the minimal genome size required for life. FEBS Lett 362:257–260PubMedCrossRefGoogle Scholar
  19. 19.
    Jay D, Gilbert W (1987) Basic protein enhances the encapsulation of DNA into lipid vesicles: model for the formation of primordial cells. PNAS 84:1978–1980PubMedCrossRefGoogle Scholar
  20. 20.
    Kolisnychenko V, Plunkett G III, Herring CD, Fehér T, Pósfai J, Blattner FR, Pósfai G (2002) Engineering a reduced Escherichia coli genome. Genome Res 12:640–647PubMedCrossRefGoogle Scholar
  21. 21.
    Koonin EV (2000) How many genes can make a cell: the minimal-gene-set concept. Annu Rev Genomics Hum Genet 1:99–116PubMedCrossRefGoogle Scholar
  22. 22.
    Koonin EV (2003) Comparative genomics, minimal gene-sets and the last universal common ancestor. Nat Rev Microbiol 1:127–136PubMedCrossRefGoogle Scholar
  23. 23.
    Koster G, Van Duijn M, Hofs B, Dogterom M (2003) Membrane tube formation from giant vesicles by dynamic association of motor proteins. PNAS 100:15583–15588PubMedCrossRefGoogle Scholar
  24. 24.
    Lazcano A, Guerriero R, Margulius L, Oró J (1988) The evolutionary transition from RNA to DNA in early cells. J Mol Evol 27:283–290PubMedCrossRefGoogle Scholar
  25. 25.
    Lazcano A, Valverde V, Hernandez G, Gariglio P, Fox GE, Oró J (1992) On the early emergence of reverse transcription: theoretical basis and experimental evidence. J Mol Evol 35:524–536PubMedCrossRefGoogle Scholar
  26. 26.
    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
  27. 27.
    Luci P (2003) Gene cloning expression and purification of membrane proteins. ETH-Z Dissertation No. 15108, Swiss Federal Institute of Technology (ETH) ZurichGoogle Scholar
  28. 28.
    Luisi PL (2002) Toward the engineering of minimal living cells. Anat Rec 268:208–214PubMedCrossRefGoogle Scholar
  29. 29.
    Luisi PL (2003) Autopoiesis: a review and a reappraisal. Naturwissenschaften 90:49–59PubMedGoogle Scholar
  30. 30.
    Luisi PL, Varela FJ (1990) Self-replicating micelles—a chemical version of minimal autopoietic systems. Orig Life Evol Biosph 19:633–643CrossRefGoogle Scholar
  31. 31.
    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
  32. 32.
    Luisi PL, Stano P, Rasi S, Mavelli F (2004) A possible route to prebiotic vesicle reproduction. Artif Life 10:297–308PubMedCrossRefGoogle Scholar
  33. 33.
    Marchi-Artzner V, Jullien L, Belloni L, Raison D, Lacombe L, Lehn JM (1996) Interaction, lipid exchange, and effect of vesicle size in systems of oppositely charged vesicles. J Phys Chem 100:13844–13856CrossRefGoogle Scholar
  34. 34.
    Monnard PA (2003) Liposome-entrapped polymerases as models for microscale/nanoscale bioreactors. J Membr Biol 191:87–97PubMedCrossRefGoogle Scholar
  35. 35.
    Morowitz HJ (1967) Biological self-replicating systems. Prog Theor Biol 1:35–58Google Scholar
  36. 36.
    Mushegian A (1999) The minimal genome concept. Curr Opin Gen Dev 9:709–714CrossRefGoogle Scholar
  37. 37.
    Mushegian A, Koonin EV (1996) A minimal gene set for cellular life derived by comparison of complete bacterial genomes. PNAS 93:10268–10273PubMedCrossRefGoogle Scholar
  38. 38.
    Nissen P, Hansen J, Ban N, Moore PB, Steitz TA (2000) The structural basis of ribosome activity in peptide bond synthesis. Science 289:920–930PubMedCrossRefGoogle Scholar
  39. 39.
    Noireaux V, Libchaber A (2004) A vesicle bioreactor as a step toward an artificial cell assembly. PNAS 101:17669–17674PubMedCrossRefGoogle Scholar
  40. 40.
    Noireaux V, Bar-Ziv R, Libchaber A (2003) Principles of cell-free genetic circuit assembly. PNAS 100:12672–12677PubMedCrossRefGoogle Scholar
  41. 41.
    Nomura SM, Tsumoto K, Hamada T, Akiyoshi K, Nakatani Y, Yoshikawa K (2003) Gene expression within cell-sized lipid vesicles. ChemBioChem 4:1172–1175PubMedCrossRefGoogle Scholar
  42. 42.
    Oberholzer T, Luisi PL (2002) The use of liposomes for constructing cell models. J Biol Phys 28:733–744CrossRefGoogle Scholar
  43. 43.
    Oberholzer T, Albrizio M, Luisi PL (1995) Polymerase chain reaction in liposomes. Chem Biol 2:677–682PubMedCrossRefGoogle Scholar
  44. 44.
    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 Commun 207:250–257PubMedCrossRefGoogle Scholar
  45. 45.
    Oberholzer T, Nierhaus KH, Luisi PL (1999) Protein expression in liposomes. Biochem Biophys Res Commun 261:238–241PubMedCrossRefGoogle Scholar
  46. 46.
    Ono N, Ikegami T (2000) Self-maintenance and self-reproduction in an abstract cell model. J Theor Biol 206:243–253PubMedCrossRefGoogle Scholar
  47. 47.
    Pantazatos DP, MacDonald RC (1999) Directly observed membrane fusion between oppositely charged phospholipid bilayers. J Membr Biol 170:27–38PubMedCrossRefGoogle Scholar
  48. 48.
    Paul N, Joyce GF (2002) A self-replicating ligase ribozyme. PNAS 99:12733–12740PubMedCrossRefGoogle Scholar
  49. 49.
    Pietrini AV, Luisi PL (2004) Cell-free protein synthesis through solubilisate exchange in water/oil emulsion compartments. ChemBioChem 5:1055–1062PubMedCrossRefGoogle Scholar
  50. 50.
    Pohorille A, Deamer D (2002) Artificial cells: prospects for biotechnology. Trends Biotechnol 20:123–128PubMedCrossRefGoogle Scholar
  51. 51.
    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
  52. 52.
    Roux A, Cappello G, Cartaud J, Prost J, Goud B, Bassereau P (2002) A minimal system allowing tubulation with molecular motors pulling on giant liposomes. PNAS 99:5394–5399PubMedCrossRefGoogle Scholar
  53. 53.
    Sankararaman S, Menon GI, Kumar PB (2004) Self-organized pattern formation in motor-microtubule mixtures. Phys Rev E 70:031905. DOI 10.1103/PhysRevE.70.031905CrossRefGoogle Scholar
  54. 54.
    Schmidli PK, Schurtenberger P, Luisi PL (1991) Liposome-mediated enzymatic synthesis of phosphatidylcholine as an approach to self-replicating liposomes. J Am Chem Soc 113:8127–8130CrossRefGoogle Scholar
  55. 55.
    Shimkets LJ (1998) Structure and sizes of genomes of the Archaea and Bacteria. In: De Bruijn FJ, Lupskin JR, Weinstock GM (eds) Bacterial genomes: physical structure and analysis. Kluwer, Boston, MA, pp 5–11Google Scholar
  56. 56.
    Spirin A (1986) Ribosome structure and protein synthesis. Benjamin Cummings, Menlo Park, CAGoogle Scholar
  57. 57.
    Stamatatos L, Leventis R, Zuckermann MJ, Silvius JR (1988) Interactions of cationic lipid vesicles with negatively charged phospholipid vesicles and biological membranes. Biochemistry 27:3917–3925PubMedCrossRefGoogle Scholar
  58. 58.
    Suttle DP, Ravel JM (1974) The effects of initiation factor 3 on the formation of 30S initiation complexes with synthetic and natural messengers. Biochem Biophys Res Commun 57:386–393PubMedCrossRefGoogle Scholar
  59. 59.
    Szathmáry E (2005) Life: in search of the simplest cell. Nature 433:469–470. DOI 10.1038/433469aPubMedCrossRefGoogle Scholar
  60. 60.
    Szostak JW, Bartel DP, Luisi PL (2001) Synthesizing life. Nature 409:387–390PubMedCrossRefGoogle Scholar
  61. 61.
    Thomas CF, Luisi PL (2004) Novel properties of DDAB: matrix effect and interaction with oleate. J Phys Chem B 108:11285–11290CrossRefGoogle Scholar
  62. 62.
    Tsumoto K, Nomura SM, Nakatani Y, Yoshikawa K (2001) Giant liposome as a biochemical reactor: transcription of DNA and transportation by laser tweezers. Langmuir 17:7225–7228CrossRefGoogle Scholar
  63. 63.
    Varela F, Maturana HR, Uribe RB (1974) Autopoiesis: the organization of living system, its characterization and a model. Biosystems 5:187–196CrossRefGoogle Scholar
  64. 64.
    Walde P, Goto A, Monnard PA, Wessicken M, Luisi PL (1994) Oparin's reactions revisited: enzymatic synthesis of poly(adenylic acid) in micelles and self-reproducing vesicles. J Am Chem Soc 116:7541–7544CrossRefGoogle Scholar
  65. 65.
    Walde P, Wick R, Fresta M, Mangone A, Luisi PL (1994) Autopoietic self-reproduction of fatty acid vesicles. J Am Chem Soc 116:11649–11654CrossRefGoogle Scholar
  66. 66.
    Weiner AM, Maizels N (1987) tRNA-like structures tag the 3′ ends of genomic RNA molecules for replication: implications for the origin of protein synthesis. PNAS 84:7383–7387PubMedCrossRefGoogle Scholar
  67. 67.
    Woese CR (1983) The primary lines of descent and the universal ancestor. In: Bendall DS (ed) Evolution from molecules to man. Cambridge University Press, Cambridge, pp 209–233Google Scholar
  68. 68.
    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 92:590–593PubMedCrossRefGoogle Scholar
  69. 69.
    Zhang B, Cech TR (1998) Peptidyl-transferase ribozymes: trans reactions, structural characterization and ribosomal RNA-like features. Chem Biol 5:539–553PubMedCrossRefGoogle Scholar
  70. 70.
    Zimmer C (2003) Tinker, tailor: can Venter stitch together a genome from scratch? Science 299:1006–1007PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Pier Luigi Luisi
    • 1
  • Francesca Ferri
    • 2
  • Pasquale Stano
    • 1
    • 3
  1. 1.Biology DepartmentUniversity of RomaTreRomeItaly
  2. 2.Biochemistry DepartmentUniversity of BolognaBolognaItaly
  3. 3.Centro Studi ‘E. Fermi’RomeItaly

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