TY  - JOUR
A1  - Okuda, Takumi
A1  - Lenz, Ann-Kathrin
A1  - Seitz, Florian
A1  - Vogel, Jörg
A1  - Höbartner, Claudia
T1  - A SAM analogue-utilizing ribozyme for site-specific RNA alkylation in living cells
T2  - Nature Chemistry
N2  - 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.
KW  - Alkyltransferase Ribozyme SAMURI
KW  - Site-specific RNA labelling
KW  - bioorthogonal SAM analogue ProSeDMA
KW  - Chemical modification
KW  - RNA
Y1  - 2023
UR  - https://opus.bibliothek.uni-wuerzburg.de/frontdoor/index/index/docId/32876
UR  - https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-328762
ER  -