Gene Expression within Cell‐Sized Lipid Vesicles

Functional protein synthesis was observed in cell‐sized lipid vesicles following encapsulation of a gene‐expression system. Expression of rsGFP (red‐shifted green fluorescent protein) within individual vesicles was observed by fluorescence microscopy. Interestingly, at the early stage of the reaction, the expression efficiency inside the vesicle was remarkably higher than that in the solution outside. The synthesized rsGFP in individual vesicles is safe from attack by proteinase K added to the external aqueous solution. Studies on cell‐sized vesicles expressing protein should contribute to a fundamental understanding of certain aspects of living systems and will be useful for practical applications, such as the construction of microreactors.

[1]  G. Zubay,et al.  In vitro synthesis of protein in microbial systems. , 1973, Annual review of genetics.

[2]  P. Luisi,et al.  Lipid vesicles as possible intermediates in the origin of life , 1999 .

[3]  S. Ichikawa,et al.  Enzymes inside lipid vesicles: preparation, reactivity and applications. , 2001, Biomolecular engineering.

[4]  P. Luisi,et al.  Polymerase chain reaction in liposomes. , 1995, Chemistry & biology.

[5]  G. Ourisson,et al.  Towards Proto‐Cells: “Primitive” Lipid Vesicles Encapsulating Giant DNA and Its Histone Complex , 2001, Cellular & molecular biology letters.

[6]  K. Yoshikawa,et al.  Spontaneous Formation of Giant Liposomes from Neutral Phospholipids , 1997 .

[7]  D. Bartel,et al.  Synthesizing life : Paths to unforeseeable science & technology , 2001 .

[8]  T. Oberholzer,et al.  Giant Vesicles as Microreactors for Enzymatic mRNA Synthesis , 2002, Chembiochem : a European journal of chemical biology.

[9]  M Wakabayashi,et al.  Synthesis of functional protein in liposome. , 2001, Journal of bioscience and bioengineering.

[10]  Kenichi Yoshikawa,et al.  Giant Liposome as a Biochemical Reactor: Transcription of DNA and Transportation by Laser Tweezers , 2001 .

[11]  H. Itoh,et al.  Preparation of giant liposomes in physiological conditions and their characterization under an optical microscope. , 1996, Biophysical journal.

[12]  Hirokazu Hotani,et al.  Giant liposomes: from membrane dynamics to cell morphogenesis , 1999 .

[13]  P. Luisi,et al.  Enzymatic RNA replication in self-reproducing vesicles: an approach to a minimal cell. , 1995, Biochemical and biophysical research communications.

[14]  G Ourisson,et al.  The terpenoid theory of the origin of cellular life: the evolution of terpenoids to cholesterol. , 1994, Chemistry & biology.

[15]  F. Menger,et al.  Chemistry and physics of giant vesicles as biomembrane models. , 1998, Current opinion in chemical biology.

[16]  P. Luisi,et al.  Protein expression in liposomes. , 1999, Biochemical and biophysical research communications.