Nucleic Acid Isotope Labelled coupled Mass Spectrometry
To bypass this problem our lab uses nucleic acid isotope labeling coupled mass spectrometry, short NAIL-MS. This approach uses stable isotope labeling of RNA nucleosides, followed by RNA purification (e.g. tRNA, rRNA, mRNA) for absolute quantification by high sensitivity mass spectrometry. This enables the discrimination of RNA modification from new transcripts and modifications from already existing ones in vivo. Moreover this approach allows multiplexing of samples, the discovery of new modifications and the absolute quantification by the use of a stable isotope labeled internal standard (SILIS). Additionally, our present tools are aimed to be extended by establishing a method to analyze RNA modifications on the oligonucleotide level by a modified version of our established nucleoside based NAIL-MS method.
It is the goal of our research to extend and apply this new tool, to examine the various mechanisms and functions of RNA modification biology in various cell types and consequently shed light on the role of RNA modifications in cell homeostasis and neurological disease pathogenesis.
