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2019 Asadi-Atoi et al.

2019 Asadi-Atoi et al.

Benefits of stable isotope labeling in RNA analysis. Biological Chemistry (2019.
Asadi-Atoi et al., 2019

Abstract

RNAs are key players in life as they connect the genetic code (DNA) with all cellular processes dominated by proteins. They contain a variety of chemical modifications and many RNAs fold into complex structures. Here, we review recent progress in the analysis of RNA modification and structure on the basis of stable isotope labeling techniques. Mass spectrometry (MS) and NMR spectroscopy are the key tools and many break-through developments were made possible by analysis of stable isotope labeled RNA. Therefore, we discuss current stable isotope labeling techniques such as metabolic labeling, enzymatic labeling and chemical synthesis. RNA structure analysis by NMR is challenging due to two major problems that become even more salient when the size of the RNA increases, namely chemical shift overlaps and line broadening leading to complete signal loss. Several isotope labeling strategies have been developed to provide solutions to these major issues, such as deuteration, segmental isotope labeling or site-specific labeling. Quantification of modified nucleosides in RNA by mass spectrometry is only possible through the application of stable isotope labeled internal standard. With nucleic acid isotope labeling coupled mass spectrometry, NAIL-MS, it is now possible to analyze the dynamic processes of post- transcriptional RNA modification and demodification. The trend, in both NMR and MS RNA analytics, is without doubt shifting from the analysis of snapshot moments towards the development and application of tools capable of analyzing the dynamics of RNA structure and modification profiles.

Publish on : 08/04/2019

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