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Monday, May 9, 2016

Real Time Gene Expression

Biochemists Watch Gene Expression in Real Time - Study | May 7, 2016

The fundamental rule of molecular biology is “DNA makes RNA makes protein,” but up until now biochemists haven’t “seen” it in action, as Albert Einstein College scientists did by adding codes of fluorescent proteins into live cells and neurons.
Albert Einstein College scientists were trying to overcome two challenges – finding a way to visualize single molecules of messenger RNA, or mRNA, as well as single molecules of protein translated from mRNA. Proteins perform most cellular functions and are the reason we are all alive.

"Using their custom microscope, Stasevich et al. have imaged RNA translation into proteins in real time. The red dots are RNA, and the proteins fluoresce blue or green. In the background the cell nucleus can be seen in green, where the mature proteins gather after they're created in the cell cytoplasm." Source;

<more at; related articles and links: (No longer lost in translation: Biochemists watch gene expression in real time. May 5, 2016) and (Real-time quantification of single RNA translation dynamics in living cells. Tatsuya Morisaki, Kenneth Lyon, Keith F. DeLuca, Jennifer G. DeLuca, Brian P. English, Zhengjian Zhang, Luke D. Lavis, Jonathan B. Grimm, Sarada Viswanathan, Loren L. Looger, Timothee Lionnet, and Timothy J. Stasevich. Science, May 2016 DOI: 10.1126/science.aaf0899. [Abstract: Although mRNA translation is a fundamental biological process, it has never been imaged in real-time with single molecule precision in vivo. To achieve this, we developed Nascent Chain Tracking (NCT), a technique that uses multi-epitope tags and antibody-based fluorescent probes to quantify single mRNA protein synthesis dynamics. NCT reveals an elongation rate of ~10 amino acids per second, with initiation occurring stochastically every ~30 s. Polysomes contain ~1 ribosome every 200-900 nucleotides and are globular rather than elongated in shape. By developing multi-color probes, we show most polysomes act independently; however, a small fraction (~5%) form complexes in which two distinct mRNAs can be translated simultaneously. The sensitivity and versatility of NCT make it a powerful new tool for quantifying mRNA translation kinetics.])>

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