Site-specific one-pot dual labeling of DNA by orthogonal cycloaddition chemistry

• Verfasst von: Schoch, Juliane [VerfasserIn]
• Verfasst von: Staudt, Markus [VerfasserIn]
• Verfasst von: Samanta, Ayan [VerfasserIn]
• Verfasst von: Wießler, Manfred [VerfasserIn]
• Verfasst von: Jäschke, Andres [VerfasserIn]
• Titel: Site-specific one-pot dual labeling of DNA by orthogonal cycloaddition chemistry
• Verf.angabe: Juliane Schoch, Markus Staudt, Ayan Samanta, Manfred Wiessler, and Andres Jäschke
• Jahr: 2012
• Umfang: 5 S.
• Fussnoten: Gesehen am 05.07.2018
• Titel Quelle: Enthalten in: Bioconjugate chemistry
• Ort Quelle: Columbus, Ohio : American Chemical Society, 1990
• Jahr Quelle: 2012
• Band/Heft Quelle: 23(2012), 7, Seite 1382-1386
• ISSN Quelle: 1520-4812
• DOI: doi:10.1021/bc300181n
• Datenträger: Online-Ressource
• Sprache: eng
• Bibliogr. Hinweis: Erscheint auch als : Druck-Ausgabe: Site-specific one-pot dual labeling of DNA by orthogonal cycloaddition chemistry. - 2012
• K10plus-PPN: 1577323394

Abstract

Bioorthogonal reactions are of high interest in biosciences as they allow the introduction of fluorescent dyes, affinity tags, or other unnatural moieties into biomolecules. The site-specific attachment of two or more different labels is particularly demanding and typically requires laborious multistep syntheses. Here, we report that the most popular cycloaddition in bioconjugation, the copper-catalyzed azide-alkyne click reaction (CuAAC), is fully orthogonal to the inverse electron-demand Diels-Alder reaction (DAinv). We demonstrate that both bioorthogonal reactions can be conducted concurrently in a one-pot reaction by just mixing all components. Orthogonality has been established even for highly reactive trans-cyclooctene-based dienophiles (with rate constants up to 380 000 M-1 s-1). These properties allow for the convenient site-specific one-step preparation of oligonucleotide FRET probes and related reporters needed in cellular biology and biophysical chemistry.