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  • Title: Site-specific chemical labeling of long RNA molecules.
    Author: Jahn K, Olsen EM, Nielsen MM, Tørring T, MohammadZadegan R, Andersen ES, Gothelf KV, Kjems J.
    Journal: Bioconjug Chem; 2011 Jan 19; 22(1):95-100. PubMed ID: 21138255.
    Abstract:
    Site-specific labeling of RNA molecules is a valuable tool for studying their structure and function. Here, we describe a new site-specific RNA labeling method, which utilizes a DNA-templated chemical reaction to attach a label at a specific internal nucleotide in an RNA molecule. The method is nonenzymatic and based on the formation of a four-way junction, where a donor strand is chemically coupled to an acceptor strand at a specific position via an activated chemical group. A disulfide bond in the linker is subsequently cleaved under mild conditions leaving a thiol group attached to the acceptor-RNA strand. The site-specific thiol-modified target RNA can then be chemically labeled with an optional group, here demonstrated by coupling of a maleimide-functionalized fluorophore. The method is rapid and allows site specific labeling of both in vitro and in vivo synthesized RNA with a broad range of functional groups.
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