Researchers developed a fluorescent microscopy method to measure mRNA degradation in single cells. The fluorescent bio-sensor allows the distinction of intact transcripts and degradation intermediates. This new method called as TREAT.
The life of an RNA starts in the nucleus, where it synthesizes and further modified. And then moves into the cytoplasm, it translates into a protein, and eventually degraded.
The level of protein produced depends on the regulation of transcription, but also the abundance of mRNA. Jeff Chao, Junior group leader at the FMI, said, the method clears the regulation of mRNA degradation, particularly during development or rapid environmental changes, can dramatically influence RNA levels.
While, the RNA degradation steps characterized, a clear understanding of when and where degradation happens has missing so far.
However, the new method measures the spatial and the temporal dynamics of the degradation of single mRNA molecules in living cells.
The TREAT technique allows scientist to observe mRNA degradation in real-time in living cells. To visualize degradation, the scientists engineered a transcript that is labeled with two RNA-binding proteins fused to two distinct fluorescent tags, PP7 and MS2. Between PP7 and MS2, the scientists introduced the viral pseudo-knots.
Using TREAT, we measure mRNA decay in single cells and found that individual degradation events occur independently in the cytosol. The degraded mRNAs did not gather in processing bodies, and also play a direct role in RNA degradation.
Processing bodies are membrane-less compartments that form during phase transitions. They comprises RNA-binding proteins and mRNAs. Since they contain many proteins involved in mRNA turnover, it proposed the cellular sites of RNA degradation.
TREAT nicely complements the other technology called TRICK. With the selection of appropriate fluorescent markers, researchers now monitor both RNA turnover and RNA translation ‘live’, and address the processes interconnected within a single cell.
More information: [Molecular cell]