Impact of Methods on the Measurement of mRNA Turnover

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,...

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Bibliographic Details
Published in:International journal of molecular sciences 2017-12, Vol.18 (12), p.2723
Main Authors: Wada, Takeo, Becskei, Attila
Format: Article
Language:English
Subjects:
4-thiouracil
4sU
Biodegradation
Equilibrium
Experiments
exponential decay
Gene expression
Half-life
Labeling
Machinery and equipment
Measurement methods
Metabolism
Methods
Molecular biology
mRNA stability
mRNA turnover
Mutation
NMD
nonsense mediated decay
posttranscriptional regulation
Quantitative analysis
Review
Ribonuclease
RNA polymerase
rpb1-1
Saccharomyces cerevisiae
splicing
Stability analysis
Transcription
Yeast
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Summary: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.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms18122723