A SAM analogue-utilizing ribozyme for site-specific RNA alkylation in living cells.
Journal
Nature chemistry
ISSN: 1755-4349
Titre abrégé: Nat Chem
Pays: England
ID NLM: 101499734
Informations de publication
Date de publication:
Nov 2023
Nov 2023
Historique:
received:
26
08
2022
accepted:
08
08
2023
medline:
6
11
2023
pubmed:
5
9
2023
entrez:
4
9
2023
Statut:
ppublish
Résumé
Post-transcriptional RNA modification methods are in high demand for site-specific RNA labelling and analysis of RNA functions. In vitro-selected ribozymes are attractive tools for RNA research and have the potential to overcome some of the limitations of chemoenzymatic approaches with repurposed methyltransferases. Here we report an alkyltransferase ribozyme that uses a synthetic, stabilized S-adenosylmethionine (SAM) analogue and catalyses the transfer of a propargyl group to a specific adenosine in the target RNA. Almost quantitative conversion was achieved within 1 h under a wide range of reaction conditions in vitro, including physiological magnesium ion concentrations. A genetically encoded version of the SAM analogue-utilizing ribozyme (SAMURI) was expressed in HEK293T cells, and intracellular propargylation of the target adenosine was confirmed by specific fluorescent labelling. SAMURI is a general tool for the site-specific installation of the smallest tag for azide-alkyne click chemistry, which can be further functionalized with fluorophores, affinity tags or other functional probes.
Identifiants
pubmed: 37667013
doi: 10.1038/s41557-023-01320-z
pii: 10.1038/s41557-023-01320-z
pmc: PMC10624628
doi:
Substances chimiques
RNA
63231-63-0
RNA, Catalytic
0
S-Adenosylmethionine
7LP2MPO46S
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
1523-1531Informations de copyright
© 2023. The Author(s).
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