A multifunctional bioconjugate module for versatile photoaffinity labeling and click chemistry of RNA

• Verfasst von: Kellner, Stefanie [VerfasserIn]
• Verfasst von: Seidu-Larry, Salifu [VerfasserIn]
• Verfasst von: Burhenne, Jürgen [VerfasserIn]
• Verfasst von: Motorin, Yuri [VerfasserIn]
• Verfasst von: Helm, Mark [VerfasserIn]
• Titel: A multifunctional bioconjugate module for versatile photoaffinity labeling and click chemistry of RNA
• Verf.angabe: Stefanie Kellner, Salifu Seidu-Larry, Jürgen Burhenne, Yuri Motorin and Mark Helm
• E-Jahr: 2011
• Jahr: 6 June 2011
• Umfang: 13 S.
• Fussnoten: Gesehen am 28.07.2022
• Titel Quelle: Enthalten in: Nucleic acids research
• Ort Quelle: Oxford : Oxford Univ. Press, 1974
• Jahr Quelle: 2011
• Band/Heft Quelle: 39(2011), 16, Seite 7348-7360
• ISSN Quelle: 1362-4962
• DOI: doi:10.1093/nar/gkr449
• Datenträger: Online-Ressource
• Sprache: eng
• K10plus-PPN: 1811903878

Abstract

A multifunctional reagent based on a coumarin scaffold was developed for derivatization of naive RNA. The alkylating agent N3BC [7-azido-4-(bromomethyl)coumarin], obtained by Pechmann condensation, is selective for uridine. N3BC and its RNA conjugates are pre-fluorophores which permits controlled modular and stepwise RNA derivatization. The success of RNA alkylation by N3BC can be monitored by photolysis of the azido moiety, which generates a coumarin fluorophore that can be excited with UV light of 320 nm. The azidocoumarin-modified RNA can be flexibly employed in structure-function studies. Versatile applications include direct use in photo-crosslinking studies to cognate proteins, as demonstrated with tRNA and RNA fragments from the MS2 phage and the HIV genome. Alternatively, the azide function can be used for further derivatization by click-chemistry. This allows e.g. the introduction of an additional fluorophore for excitation with visible light.