Quantitative analysis of m(6)A RNA modification by LC-MS

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)...

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Detalles Bibliográficos
Autores principales: Mathur, Lavina, Jung, Sunhee, Jang, Cholsoon, Lee, Gina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8353476/
https://www.ncbi.nlm.nih.gov/pubmed/34401789
http://dx.doi.org/10.1016/j.xpro.2021.100724
Descripción
Sumario: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).

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