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September 2016-March 2017: Master Thesis on the dynamics of RNA modifications in S. cerevisiae.
Since June 2017: PhD thesis on the dynamics of RNA modifications in neurological disease.
Investigation of RNA modification dynamics in neurological disease by NAIL-MS
RNA modifications are involved in many molecular mechanisms such as translation fidelity or folding of tertiary structures. Deviations of the “normal” modification pattern inside a cell most often results in diverse neurological diseases.
So far, it was challenging to investigate these modification dynamics, as most methods only display a static view. With our developed NAIL-MS approach (nucleic acid isotope labeling coupled mass spectrometry) we want to get a deeper insight into modification dynamics in several organisms.
I am particularly interested in the investigation of modification dynamics in human cell culture. Knockout strains or multiple stressors are used to see how the cells react to environmental challenges. Therefore I use NAIL-MS: The general workflow consist of heavy isotope labeling of RNA nucleosides by feeding precursor molecules of nucleoside biosynthesis. After total RNA isolation and purification of the desired RNA species by size exclusion chromatography (e.g. tRNA, rRNA), the RNA is digested to nucleosides and finally analyzed by high sensitivity mass spectrometry. This allows the discrimination of isotopomers and thereby enables the elucidation of many RNA modification mechanisms inside the cell.