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
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author Mathur, Lavina
Jung, Sunhee
Jang, Cholsoon
Lee, Gina
author_facet Mathur, Lavina
Jung, Sunhee
Jang, Cholsoon
Lee, Gina
author_sort Mathur, Lavina
collection PubMed
description 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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spelling pubmed-83534762021-08-15 Quantitative analysis of m(6)A RNA modification by LC-MS Mathur, Lavina Jung, Sunhee Jang, Cholsoon Lee, Gina STAR Protoc Protocol 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). Elsevier 2021-08-06 /pmc/articles/PMC8353476/ /pubmed/34401789 http://dx.doi.org/10.1016/j.xpro.2021.100724 Text en © 2021 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Mathur, Lavina
Jung, Sunhee
Jang, Cholsoon
Lee, Gina
Quantitative analysis of m(6)A RNA modification by LC-MS
title Quantitative analysis of m(6)A RNA modification by LC-MS
title_full Quantitative analysis of m(6)A RNA modification by LC-MS
title_fullStr Quantitative analysis of m(6)A RNA modification by LC-MS
title_full_unstemmed Quantitative analysis of m(6)A RNA modification by LC-MS
title_short Quantitative analysis of m(6)A RNA modification by LC-MS
title_sort quantitative analysis of m(6)a rna modification by lc-ms
topic Protocol
url 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
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