In:
Angewandte Chemie, Wiley, Vol. 134, No. 49 ( 2022-12-05)
Abstract:
5‐Formylcytidine (f 5 C) is one of the epigenetic nucleotides in tRNA. Despite the evident importance of f 5 C in gene expression regulation, little is known about its exact amount and position. To capture this information, we developed a modification‐specific functionalization with a semi‐stabilized ylide. The chemical labelling exhibited a high selectivity towards f 5 C and allowed distinction from similar 5‐formyluridine. We realized a detection strategy based on the fluorescence signal of the cyclization product 4,5‐pyridin‐2‐amine‐cytidine paC, which exhibited a high quantum yield. The results clearly identified f 5 C with a limit of detection at 0.58 nM. This method altered the hydrogen bonding activities of f 5 C and modulated its reverse transcription signature in its sequencing profile. We showed that f 5 C can be detected from tRNA segments with a single‐base resolution. Taken together, this approach is a sensitive, antibody‐free, and applicable detection and sequencing method for f 5 C‐containing RNA.
Type of Medium:
Online Resource
ISSN:
0044-8249
,
1521-3757
DOI:
10.1002/ange.v134.49
DOI:
10.1002/ange.202210652
Language:
English
Publisher:
Wiley
Publication Date:
2022
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