Detection of m(6)A RNA modifications at single-nucleotide resolution using m(6)A-selective allyl chemical labeling and sequencing

As the most abundant internal mRNA modification, N(6)-methyladenosine (m(6)A) was involved in almost all the aspects of RNA metabolism. Here, we introduce our protocol for m(6)A-SAC-seq, which enables the whole transcriptome-wide mapping of m(6)A RNA modification at single-nucleotide resolution with...

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Autores principales: Peng, Yong, Meng, Hanzhe, Ge, Ruiqi, Liu, Shun, Chen, Mengjie, He, Chuan, Hu, Lulu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9485529/
https://www.ncbi.nlm.nih.gov/pubmed/36112507
http://dx.doi.org/10.1016/j.xpro.2022.101677
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author Peng, Yong
Meng, Hanzhe
Ge, Ruiqi
Liu, Shun
Chen, Mengjie
He, Chuan
Hu, Lulu
author_facet Peng, Yong
Meng, Hanzhe
Ge, Ruiqi
Liu, Shun
Chen, Mengjie
He, Chuan
Hu, Lulu
author_sort Peng, Yong
collection PubMed
description As the most abundant internal mRNA modification, N(6)-methyladenosine (m(6)A) was involved in almost all the aspects of RNA metabolism. Here, we introduce our protocol for m(6)A-SAC-seq, which enables the whole transcriptome-wide mapping of m(6)A RNA modification at single-nucleotide resolution with stoichiometry information. m(6)A-SAC-seq relies on selective allyl labeling of m(6)A by specific methyltransferase and chemical treatment that introduce mutation upon reverse transcription. The technique only requires ∼30 ng of input RNA. For complete details on the use and execution of this protocol, please refer to Hu et al. (2022).
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spelling pubmed-94855292022-09-21 Detection of m(6)A RNA modifications at single-nucleotide resolution using m(6)A-selective allyl chemical labeling and sequencing Peng, Yong Meng, Hanzhe Ge, Ruiqi Liu, Shun Chen, Mengjie He, Chuan Hu, Lulu STAR Protoc Protocol As the most abundant internal mRNA modification, N(6)-methyladenosine (m(6)A) was involved in almost all the aspects of RNA metabolism. Here, we introduce our protocol for m(6)A-SAC-seq, which enables the whole transcriptome-wide mapping of m(6)A RNA modification at single-nucleotide resolution with stoichiometry information. m(6)A-SAC-seq relies on selective allyl labeling of m(6)A by specific methyltransferase and chemical treatment that introduce mutation upon reverse transcription. The technique only requires ∼30 ng of input RNA. For complete details on the use and execution of this protocol, please refer to Hu et al. (2022). Elsevier 2022-09-15 /pmc/articles/PMC9485529/ /pubmed/36112507 http://dx.doi.org/10.1016/j.xpro.2022.101677 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Peng, Yong
Meng, Hanzhe
Ge, Ruiqi
Liu, Shun
Chen, Mengjie
He, Chuan
Hu, Lulu
Detection of m(6)A RNA modifications at single-nucleotide resolution using m(6)A-selective allyl chemical labeling and sequencing
title Detection of m(6)A RNA modifications at single-nucleotide resolution using m(6)A-selective allyl chemical labeling and sequencing
title_full Detection of m(6)A RNA modifications at single-nucleotide resolution using m(6)A-selective allyl chemical labeling and sequencing
title_fullStr Detection of m(6)A RNA modifications at single-nucleotide resolution using m(6)A-selective allyl chemical labeling and sequencing
title_full_unstemmed Detection of m(6)A RNA modifications at single-nucleotide resolution using m(6)A-selective allyl chemical labeling and sequencing
title_short Detection of m(6)A RNA modifications at single-nucleotide resolution using m(6)A-selective allyl chemical labeling and sequencing
title_sort detection of m(6)a rna modifications at single-nucleotide resolution using m(6)a-selective allyl chemical labeling and sequencing
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9485529/
https://www.ncbi.nlm.nih.gov/pubmed/36112507
http://dx.doi.org/10.1016/j.xpro.2022.101677
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