Tracking mRNA in living cells : Analyzing the kinetics of mRNA dynamics using an MS2 based system
2025
Kraft, Aron
Translation is crucial to living cells and is itself highly dependent on the journey of mRNA.mRNA is responsible for delivering the embedded code of the DNA to the ribosome in order forthe correct proteins to be produced. The details of this system are complex and hard to graspby in vitro experiments. mRNA are translated to proteins in the cytosol and, in the case ofmembrane proteins, the protein can get inserted into the membrane as translation is going on.This single-molecule tracking project aims to further experiment with an existing MS2 system fortracking mRNA in vivo and analyze the kinetics of mRNA dynamics inside E. coli cells. Thesystem utilizes the affinity between the MS2 RNA stem loop and the MS2 bacteriophage coatprotein (MS2CP). The latter is fused to Halotag (creating MS2CPHalo) in order for tracking tobecome viable through fluorescence microscopy. We show that the system is capable oftracking membrane protein mRNA and displays key differences in mRNA behaviour underdifferent translation conditions, such as co-translational insertion and ribosome binding site(RBS) mutation.
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