AlkAniline-Seq: Profiling of m7 G and m3 C RNA Modifications at Single Nucleotide Resolution
| Autor: | Adeline Galvanin, Jasmin Hertler, Valérie Bourguignon-Igel, Yuri Motorin, Virginie Marchand, Denis L. J. Lafontaine, Felix G.M. Ernst, Lilia Ayadi, Mark Helm, Annika Kotter |
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| Přispěvatelé: | Ingénierie, Biologie et Santé en Lorraine (IBSLor), Université de Lorraine (UL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Ingénierie Moléculaire et Physiopathologie Articulaire (IMoPA), Centre National de la Recherche Scientifique (CNRS)-Université de Lorraine (UL), Université libre de Bruxelles (ULB), Center for Microscopy and Molecular Imaging (IBMM - CMMI), Johannes Gutenberg - Universität Mainz (JGU) |
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Computational biology Catalysis Deep sequencing 03 medical and health sciences deep sequencing Adapter (genetics) [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry Molecular Biology/Genomics [q-bio.GN] Epitranscriptomics abasic site Nucleotide AP site ComputingMilieux_MISCELLANEOUS chemistry.chemical_classification Regulation of gene expression Chemistry RNA [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry Molecular Biology/Molecular biology General Chemistry Methylation Sciences bio-médicales et agricoles RNA modification 3. Good health 030104 developmental biology methylation epitranscriptomics |
| Zdroj: | Angewandte Chemie, 57 (51 Angewandte Chemie International Edition Angewandte Chemie International Edition, Wiley-VCH Verlag, 2018, 57 (51), pp.16785-16790. ⟨10.1002/anie.201810946⟩ |
| ISSN: | 1433-7851 1521-3773 |
| DOI: | 10.1002/anie.201810946⟩ |
| Popis: | RNA modifications play essential roles in gene expression regulation. Only seven out of >150 known RNA modifications are detectable transcriptome-wide by deep sequencing. Here we describe a new principle of RNAseq library preparation, which relies on a chemistry based positive enrichment of reads in the resulting libraries, and therefore leads to unprecedented signal-to-noise ratios. The proposed approach eschews conventional RNA sequencing chemistry and rather exploits the generation of abasic sites and subsequent aniline cleavage. The newly generated 5'-phosphates are used as unique entry for ligation of an adapter in library preparation. This positive selection, embodied in the AlkAniline-Seq, enables a deep sequencing-based technology for the simultaneous detection of 7-methylguanosine (m7 G) and 3-methylcytidine (m3 C) in RNA at single nucleotide resolution. As a proof-of-concept, we used AlkAniline-Seq to comprehensively validate known m7 G and m3 C sites in bacterial, yeast, and human cytoplasmic and mitochondrial tRNAs and rRNAs, as well as for identifying previously unmapped positions. SCOPUS: ar.j info:eu-repo/semantics/published |
| Databáze: | OpenAIRE |
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