Unbiased tracking of the progression of mRNA and protein synthesis in bulk and in liposome-confined reactions

Pauline van Nies, Zohreh Nourian, Maurits Kok, Roeland van Wijk, Jonne Moeskops, Ilja Westerlaken, Jos M. Poolman, Rienk Eelkema, Jan H. van Esch, Yutetsu Kuruma, Takuya Ueda, Christophe Danelon

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

The compartmentalization of a cell-free gene expression system inside a self-assembled lipid vesicle is envisioned as the simplest chassis for the construction of a minimal cell. Although crucial for its realization, quantitative understanding of the dynamics of gene expression in bulk and liposome-confined reactions is scarce. Here, we used two orthogonal fluorescence labeling tools to report the amounts of mRNA and protein produced in a reconstituted biosynthesis system, simultaneously and in real-time. The Spinach RNA aptamer and its fluorogenic probe were used for mRNA detection. Applying this dual-reporter assay to the analysis of transcript and protein production inside lipid vesicles revealed that their levels are uncorrelated, most probably a consequence of the low copy-number of some components in liposome-confined reactions. We believe that the stochastic nature of gene expression should be appreciated as a design principle for the assembly of a minimal cell.

Original languageEnglish
Pages (from-to)1963-1966
Number of pages4
JournalChemBioChem
Volume14
Issue number15
DOIs
Publication statusPublished - 2013 Oct 1

Keywords

  • Artificial cells
  • Gene expression
  • Liposomes
  • RNA aptamers
  • Self-assembly

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Organic Chemistry

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    van Nies, P., Nourian, Z., Kok, M., van Wijk, R., Moeskops, J., Westerlaken, I., Poolman, J. M., Eelkema, R., van Esch, J. H., Kuruma, Y., Ueda, T., & Danelon, C. (2013). Unbiased tracking of the progression of mRNA and protein synthesis in bulk and in liposome-confined reactions. ChemBioChem, 14(15), 1963-1966. https://doi.org/10.1002/cbic.201300449