Abstract
Turnover of mRNA is an important level of gene regulation. Individual mRNAs have different intrinsic stabilities. Moreover, mRNA stability changes dynamically with conditions such as hormonal stimulation or cellular stress. While accurate methods exist to measure the half-life of an individual transcript, global methods to estimate mRNA turnover have limitations in terms of resolution in time and precision. We describe and compare two complementary approaches to estimating global transcript stability: (1) direct measurement of decay rates; (2) indirect estimation of turnover from determination of mRNA synthesis rates and steady-state levels. Since the two approaches have distinct strengths yet confer different cellular perturbations, it is valuable to consider results obtained with both methods. The practical aspects of the chapter are written from a yeast perspective; the general considerations hold true for all eukaryotes, however.
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Acknowledgments
Work in the authors’ laboratories is supported by grants from the Spanish MEC (BIO2007-67708-C04-02) and MiCInn (BFU2009-11965, BFU2008-02114, BFU2007-67575-C03-01/BMC), and by the Swedish Research Council (2007-5460).
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Appendix: The MARMOR Program
Appendix: The MARMOR Program
1.1 Macro 1
1.2 Macro 2
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Marín-Navarro, J., Jauhiainen, A., Moreno, J., Alepuz, P., Pérez-Ortín, J.E., Sunnerhagen, P. (2011). Global Estimation of mRNA Stability in Yeast. In: Becskei, A. (eds) Yeast Genetic Networks. Methods in Molecular Biology, vol 734. Humana Press. https://doi.org/10.1007/978-1-61779-086-7_1
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DOI: https://doi.org/10.1007/978-1-61779-086-7_1
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