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Impact of Methods on the Measurement of mRNA Turnover

Biozentrum, University of Basel, Klingelbergstrasse 50/70, 4056 Basel, Switzerland
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Int. J. Mol. Sci. 2017, 18(12), 2723; https://doi.org/10.3390/ijms18122723
Received: 25 October 2017 / Revised: 4 December 2017 / Accepted: 8 December 2017 / Published: 15 December 2017
(This article belongs to the Section Biochemistry)
The turnover of the RNA molecules is determined by the rates of transcription and RNA degradation. Several methods have been developed to study RNA turnover since the beginnings of molecular biology. Here we summarize the main methods to measure RNA half-life: transcription inhibition, gene control, and metabolic labelling. These methods were used to detect the cellular activity of the mRNAs degradation machinery, including the exo-ribonuclease Xrn1 and the exosome. On the other hand, the study of the differential stability of mature RNAs has been hampered by the fact that different methods have often yielded inconsistent results. Recent advances in the systematic comparison of different method variants in yeast have permitted the identification of the least invasive methodologies that reflect half-lives the most faithfully, which is expected to open the way for a consistent quantitative analysis of the determinants of mRNA stability. View Full-Text
Keywords: posttranscriptional regulation; Saccharomyces cerevisiae; nonsense mediated decay; NMD; splicing; 4-thiouracil; 4sU; rpb1-1; exponential decay posttranscriptional regulation; Saccharomyces cerevisiae; nonsense mediated decay; NMD; splicing; 4-thiouracil; 4sU; rpb1-1; exponential decay
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Wada, T.; Becskei, A. Impact of Methods on the Measurement of mRNA Turnover. Int. J. Mol. Sci. 2017, 18, 2723.

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