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m6A RNA modifications are measured at single-base resolution across the mammalian transcriptome
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m6A RNA modifications are measured at single-base resolution across the mammalian transcriptome
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Journal Article
m6A RNA modifications are measured at single-base resolution across the mammalian transcriptome
2022
Overview
Functional studies of the RNA
N
6
-methyladenosine (m
6
A) modification have been limited by an inability to map individual m
6
A-modified sites in whole transcriptomes. To enable such studies, here, we introduce m
6
A-selective allyl chemical labeling and sequencing (m
6
A-SAC-seq), a method for quantitative, whole-transcriptome mapping of m
6
A at single-nucleotide resolution. The method requires only ~30 ng of poly(A) or rRNA-depleted RNA. We mapped m
6
A modification stoichiometries in RNA from cell lines and during in vitro monocytopoiesis from human hematopoietic stem and progenitor cells (HSPCs). We identified numerous cell-state-specific m
6
A sites whose methylation status was highly dynamic during cell differentiation. We observed changes of m
6
A stoichiometry as well as expression levels of transcripts encoding or regulated by key transcriptional factors (TFs) critical for HSPC differentiation. m
6
A-SAC-seq is a quantitative method to dissect the dynamics and functional roles of m
6
A sites in diverse biological processes using limited input RNA.
m
6
A-SAC-seq uses chemical labeling to quantify m
6
A at single-base resolution in the mammalian transcriptome.
Publisher
Nature Publishing Group US,Nature Publishing Group
Subject
