Visualization of single ribonucleic acid in living cell achieved?

Yes says a research group lead by Philip Santangelo, an Asst., Professor in the Wallace H. Coulter Department of Biomedical Engineering at Georgia Tech and Emory University. This finding is of importance because of the fact that, this tool will help scientists learn more about how RNA operates within living cells. And more over the researchers have overcome some of the drawbacks of the earlier method like “the need for synthetic RNA or a large number of fluorescent molecules”.

In the study, the probes - produced by attaching a few small fluorescent molecules called fluorophores to a modified nucleic acid sequence and combining the sequences with a protein - exhibited single-molecule sensitivity and allowed the researchers to target and follow native RNA and non-engineered viral RNA in living cells.

The significance of the research lies in the fact that the probes recognize RNA sequences and bind to them using the same base pairing most of us are familiar in regards to DNA, For their experiments, the team used a bacterial toxin to transport the probes into living cells - a delivery technique that when combined with the high affinity of the probes for their targets, required significantly fewer probes than existing techniques. The toxin created several tiny holes in the cell membrane that allowed the probes to enter the cell's cytoplasm and later testing the sensitivity by the conventional fluorescence microscopy to image individual probes inside a cell. More interestingly, they were able to overcome the draw back of earlier method like “accumulation of probes inside a cell”.

With single-molecule sensitivity accomplished, the researchers investigated whether they could visualize individual RNA molecules using the probes. To do this, they simultaneously delivered probes designed to target a human messenger RNA (mRNA) sequence region and a probe designed with no target in the human genome. They were able to image unbound probes of both types as well as individual RNA molecules that had attached to the former probes.

With this the researchers also were, able to observe a process called dynamic RNA-protein co-localization (joining of RNA molecules and RNA binding proteins in a single cell). Congratulations for the group and wish them further success in their endeavor…More..