m(6)A RNA modifications are measured at single-base resolution across the mammalian transcriptome Journal Article
| Authors: | Hu, L.; Liu, S.; Peng, Y.; Ge, R.; Su, R.; Senevirathne, C.; Harada, B. T.; Dai, Q.; Wei, J.; Zhang, L.; Hao, Z.; Luo, L.; Wang, H.; Wang, Y.; Luo, M.; Chen, M.; Chen, J.; He, C. |
| Article Title: | m(6)A RNA modifications are measured at single-base resolution across the mammalian transcriptome |
| 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. © 2022, The Author(s), under exclusive licence to Springer Nature America, Inc. |
| Keywords: | methylation; genetics; animal; metabolism; animals; cell differentiation; rna; cell culture; messenger rna; rna, messenger; mammal; stem cells; alkylation; rna processing; mammals; in-vitro; transcriptome; rna processing, post-transcriptional; stoichiometry; hematopoietic progenitor cells; cell lines; chemical labeling; transcriptomes; humans; human; single nucleotides; chemical sequencing; haematopoietic stem cells; single basis |
| Journal Title: | Nature Biotechnology |
| Volume: | 40 |
| Issue: | 8 |
| ISSN: | 1087-0156 |
| Publisher: | Nature Publishing Group |
| Date Published: | 2022-08-01 |
| Start Page: | 1210 |
| End Page: | 1219 |
| Language: | English |
| DOI: | 10.1038/s41587-022-01243-z |
| PUBMED: | 35288668 |
| PROVIDER: | scopus |
| PMCID: | PMC9378555 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 3 October 2022 -- Source: Scopus |
