| Literature DB >> 35288668 |
Lulu Hu1,2,3,4, Shun Liu5,6,7,8,9, Yong Peng5,6,7,8,9, Ruiqi Ge5,6,7, Rui Su10, Chamara Senevirathne11, Bryan T Harada5,6,7, Qing Dai5,6,7, Jiangbo Wei5,6,7, Lisheng Zhang5,6,7, Ziyang Hao5,6,7, Liangzhi Luo5,6,7, Huanyu Wang5,6,7, Yuru Wang5,6,7, Minkui Luo11,12, Mengjie Chen13,14, Jianjun Chen15,16,17, Chuan He18,19,20.
Abstract
Functional studies of the RNA N6-methyladenosine (m6A) modification have been limited by an inability to map individual m6A-modified sites in whole transcriptomes. To enable such studies, here, we introduce m6A-selective allyl chemical labeling and sequencing (m6A-SAC-seq), a method for quantitative, whole-transcriptome mapping of m6A at single-nucleotide resolution. The method requires only ~30 ng of poly(A) or rRNA-depleted RNA. We mapped m6A 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 m6A sites whose methylation status was highly dynamic during cell differentiation. We observed changes of m6A stoichiometry as well as expression levels of transcripts encoding or regulated by key transcriptional factors (TFs) critical for HSPC differentiation. m6A-SAC-seq is a quantitative method to dissect the dynamics and functional roles of m6A sites in diverse biological processes using limited input RNA.Entities:
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Year: 2022 PMID： 35288668 PMCID： PMC9378555 DOI： 10.1038/s41587-022-01243-z
Source DB: PubMed Journal: Nat Biotechnol ISSN： 1087-0156 Impact factor: 68.164