A SAM Analogue-utilizing Ribozyme for Site-specific RNA Alkylation in Living Cells

Overview

Journal Nat Chem

Specialty Chemistry

Date 2023 Sep 4

PMID 37667013

Authors

Takumi Okuda

Ann-Kathrin Lenz

Florian Seitz

Jorg Vogel

Claudia Hobartner

Affiliations

Soon will be listed here.

Abstract

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.

Citing Articles

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