How can we study mRNA half-lives?
Early on, several methods or approaches were utilized for the studies of mRNA half-lives.
- Transcription Inhibitors: These methods utilized transcription inhibitors such as actinomycin, DRB, cordycepin, and α-amanitin.
- Pulse-labeling: Radioactive nucleosides are added to cells and “chased” with unlabeled nucleosides.
- Steady state approaches: Cells are continuously labeled followed by the quantification of accumulated radioactive mRNA.
- Degradation of mRNA in vitro: Several methods are available. A crude cytosol, polysomes, or messenger ribonucleoprotein for nucleated cells are prepared and the decay of cell-derived mRNA is studied.
- Decay of mRNA in animals: Animal models are utilized for this type of studies. Often, whole-genome microarrays are employed.
- Stability of mRNAs: Variables such as primary and secondary structure, translation rate, intracellular location, and others inﬂuence mRNA stability. The stability of a specific mRNA can be affected by minor changes in structure. Some changes in sequence may inﬂuence the half-life indirectly as well. Truncated mRNAs appear to be more stable than their wild-type counterparts.