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Updated: May 22, 2026
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Sequence-specific Labeling of Nucleic Acids and Proteins with Methyltransferases and Cofactor Analogues
Published on: November 22, 2014
Programmable sequence-specific click-labeling of RNA using archaeal box C/D RNP methyltransferases
Migle Tomkuviene1, Béatrice Clouet-d'Orval, Ignas Cerniauskas
1Department of Biological DNA Modification, Institute of Biotechnology, Vilnius University, Vilnius LT-02241, Lithuania.
Nucleic Acids Research
|May 9, 2012
Summary
Researchers developed a new method for precisely labeling RNA molecules with chemical probes. This technique enables sequence-specific modification of RNA, advancing biophysical and mechanistic studies of RNA function.
Area of Science:
- Biochemistry
- Molecular Biology
- RNA Biology
Background:
- Investigating RNA function requires precise labeling with spectroscopic and chemical probes.
- Current technologies face challenges in achieving site-specific RNA modification.
Purpose of the Study:
- To develop a novel method for synthetically tunable, sequence-specific RNA labeling.
- To enable site-specific incorporation of chemical probes for RNA research.
Main Methods:
- Reconstitution of a functional box C/D small ribonucleoprotein RNA 2'-O-methyltransferase (C/D RNP) from Pyrococcus abyssi.
- In vitro transfer of a prop-2-ynyl group to target sites in tRNA and pre-mRNA using a modified C/D guide RNA.
- Further functionalization of the transferred alkyne group via click chemistry.
Main Results:
- Demonstrated efficient derivatization of preselected target sites in RNA molecules.
- Showcased programmable target selection by altering the C/D guide RNA sequence.
- Successfully appended a fluorophore to the transferred alkyne using bioorthogonal click reaction.
Conclusions:
- The developed approach allows for the first time synthetically tunable, sequence-specific RNA labeling with single-nucleotide precision.
- This method significantly advances the ability to probe RNA structure and function.
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