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Multiplexed gene control reveals rapid mRNA turnover.

  • Baudrimont A Biozentrum, University of Basel, 4056 Basel, Switzerland.
  • Voegeli S Biozentrum, University of Basel, 4056 Basel, Switzerland.
  • Viloria EC Biozentrum, University of Basel, 4056 Basel, Switzerland.
  • Stritt F Biozentrum, University of Basel, 4056 Basel, Switzerland.
  • Lenon M Biozentrum, University of Basel, 4056 Basel, Switzerland.
  • Wada T Biozentrum, University of Basel, 4056 Basel, Switzerland.
  • Jaquet V Biozentrum, University of Basel, 4056 Basel, Switzerland.
  • Becskei A Biozentrum, University of Basel, 4056 Basel, Switzerland.
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  • 2017-07-15
Published in:
  • Science advances. - 2017
Biological Assay
Gene Expression
Gene Expression Regulation
Genes, Reporter
Half-Life
Kinetics
Models, Biological
Open Reading Frames
Promoter Regions, Genetic
RNA Stability
RNA, Messenger
Transcription, Genetic
English The rates of mRNA synthesis and decay determine the mRNA expression level. The two processes are under coordinated control, which makes the measurements of these rates challenging, as evidenced by the low correlation among the methods of measurement of RNA half-lives. We developed a minimally invasive method, multiplexed gene control, to shut off expression of genes with controllable synthetic promoters. The method was validated by measuring the ratios of the nascent to mature mRNA molecules and by measuring the half-life with endogenous promoters that can be controlled naturally or through inserting short sequences that impart repressibility. The measured mRNA half-lives correlated highly with those obtained with the metabolic pulse-labeling method in yeast. However, mRNA degradation was considerably faster in comparison to previous estimates, with a median half-life of around 2 min. The half-life permits the estimation of promoter-dependent and promoter-independent transcription rates. The dynamical range of the promoter-independent transcription rates was larger than that of the mRNA half-lives. The rapid mRNA turnover and the broad adjustability of promoter-independent transcription rates are expected to have a major impact on stochastic gene expression and gene network behavior.
Language
  • English
Open access status
gold
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Persistent URL
https://sonar.ch/global/documents/160222
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