m6Am RNA modification detection by m6Am-seq
Copyright © 2021 Elsevier Inc. All rights reserved..
Reversible and dynamic RNA modifications play important roles in fine-tuning gene expression. N6, 2'-O-dimethyladenosine (m6Am), a terminal modification at mRNA cap, mediates various biological effects. However, limitations of the current m6Am detection methods lead to a lack of potential applications. Here, we describe a specific and sensitive method, termed m6Am-seq, that can detect m6Am at single-base resolution. m6Am-seq is based on optimized in-vitro demethylation assay and RNA immunoprecipitation, which can distinguish m6Am from 5'-UTR m6A. We provide a step by step protocol to perform m6Am-seq, including experimental procedures and sequencing data analysis. Collectively, we describe m6Am-seq, a robust tool to reveal both m6Am and 5'-UTR m6A methylome, enabling further functional and mechanistic study of m6Am modification.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2022 |
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| Erschienen: |
2022 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:203 |
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| Enthalten in: |
Methods (San Diego, Calif.) - 203(2022) vom: 01. Juli, Seite 242-248 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Zhang, Meiling [VerfasserIn] |
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| Links: |
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| Themen: |
63231-63-0 |
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| Anmerkungen: |
Date Completed 21.06.2022 Date Revised 27.06.2022 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.ymeth.2021.10.001 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM331654679 |
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| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2021 Elsevier Inc. All rights reserved. | ||
| 520 | |a Reversible and dynamic RNA modifications play important roles in fine-tuning gene expression. N6, 2'-O-dimethyladenosine (m6Am), a terminal modification at mRNA cap, mediates various biological effects. However, limitations of the current m6Am detection methods lead to a lack of potential applications. Here, we describe a specific and sensitive method, termed m6Am-seq, that can detect m6Am at single-base resolution. m6Am-seq is based on optimized in-vitro demethylation assay and RNA immunoprecipitation, which can distinguish m6Am from 5'-UTR m6A. We provide a step by step protocol to perform m6Am-seq, including experimental procedures and sequencing data analysis. Collectively, we describe m6Am-seq, a robust tool to reveal both m6Am and 5'-UTR m6A methylome, enabling further functional and mechanistic study of m6Am modification | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Demethylation | |
| 650 | 4 | |a RNA immunoprecipitation | |
| 650 | 4 | |a Single-base resolution | |
| 650 | 4 | |a m(6)Am | |
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| 650 | 7 | |a Adenosine |2 NLM | |
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| 700 | 1 | |a Sun, Hanxiao |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Kai |e verfasserin |4 aut | |
| 700 | 1 | |a Xiao, Ye |e verfasserin |4 aut | |
| 700 | 1 | |a Yi, Chengqi |e verfasserin |4 aut | |
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