Site-Specific Chemical Labeling of Long RNA Molecules

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  • RNA

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  • Molekylærbiologisk Institut
  • Interdisciplinær Nanoscience Forskerskole
  • Institut for Kemi
  • Interdisciplinary Nanoscience Center
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.
OriginalsprogEngelsk
TidsskriftBioconjugate Chemistry
Vol/bind22
Nummer1
Sider (fra-til)95-100
Antal sider6
ISSN1043-1802
DOI
StatusUdgivet - 19 jan. 2011

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