Quantitative analysis of m6A RNA modification by LC-MS
Lavina Mathur, Sunhee Jung, Cholsoon Jang, Gina Lee
Abstract
N 6 -adenosine methylation (m 6 A) of messenger RNA (mRNA) plays key regulatory roles in gene expression. Accurate measurement of m 6 A levels is thus critical to understand its dynamic changes in various biological settings. Here, we provide a protocol to quantitate the levels of adenosine and m 6 A in cellular mRNAs. Using nuclease and phosphatase, we digest mRNA into nucleosides, which are subsequently quantified using liquid chromatography mass spectrometry. For complete details on the use and execution of this protocol, please refer to Cho et al. (2021).
Topics & Concepts
RNAMessenger RNANucleaseNuclease protection assayGene expressionChemistryAdenosineN6-MethyladenosineMolecular biologyMethylationBiochemistryCell biologyGeneBiologyNon-coding RNAMethyltransferaseRNA modifications and cancerCancer-related gene regulationRNA and protein synthesis mechanisms