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Detection of m⁶A RNA modifications at single-nucleotide resolution using m⁶A-selective allyl chemical labeling and sequencing.

ABSTRACT: As the most abundant internal mRNA modification, N⁶-methyladenosine (m⁶A) was involved in almost all the aspects of RNA metabolism. Here, we introduce our protocol for m⁶A-SAC-seq, which enables the whole transcriptome-wide mapping of m⁶A RNA modification at single-nucleotide resolution with stoichiometry information. m⁶A-SAC-seq relies on selective allyl labeling of m⁶A 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).

SUBMITTER: Peng Y

PROVIDER: S-EPMC9485529 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature

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Detection of m6A RNA modifications at single-nucleotide resolution using m6A-selective allyl chemical labeling and sequencing.

Peng Yong Y Meng Hanzhe H Ge Ruiqi R Liu Shun S Chen Mengjie M He Chuan C Hu Lulu L

STAR protocols 20220915 4

As the most abundant internal mRNA modification, N6-methyladenosine (m6A) was involved in almost all the aspects of RNA metabolism. Here, we introduce our protocol for m6A-SAC-seq, which enables the whole transcriptome-wide mapping of m6A RNA modification at single-nucleotide resolution with stoichiometry information. m6A-SAC-seq relies on selective allyl labeling of m6A by specific methyltransferase and chemical treatment that in ...[more]

PMID: 36112507

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