期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 59, 期 17, 页码 6881-6886出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201916272
关键词
gene expression; nucleoside modifications; oligonucleotides; RNA sequencing; RNA structures
资金
- Austrian Science Fund FWF [P27947, P31691, F8011-B, P27024-BBL, F8009-B]
- Austrian Research Promotion Agency FFG [West Austrian BioNMR] [858017]
- Wiener Wissenschafts-, Forschungs- und Technologiefonds [WWTF LS17-003]
- Austrian Science Fund (FWF) [P27024] Funding Source: Austrian Science Fund (FWF)
Temporal information about cellular RNA populations is essential to understand the functional roles of RNA. We have developed the hydrazine/NH4Cl/OsO4-based conversion of 6-thioguanosine (6sG) into A ', where A ' constitutes a 6-hydrazino purine derivative. A ' retains the Watson-Crick base-pair mode and is efficiently decoded as adenosine in primer extension assays and in RNA sequencing. Because 6sG is applicable to metabolic labeling of freshly synthesized RNA and because the conversion chemistry is fully compatible with the conversion of the frequently used metabolic label 4-thiouridine (4sU) into C, the combination of both modified nucleosides in dual-labeling setups enables high accuracy measurements of RNA decay. This approach, termed TUC-seq DUAL, uses the two modified nucleosides in subsequent pulses and their simultaneous detection, enabling mRNA-lifetime evaluation with unprecedented precision.
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