Fast and Efficient Postsynthetic DNA Labeling in Cells by Means of Strain-Promoted Sydnone-Alkyne Cycloadditions |
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Authors: | Dr. Katja Krell Bastian Pfeuffer Dr. Franziska Rönicke Dr. Zoeisha S. Chinoy Dr. Camille Favre Dr. Frédéric Friscourt Prof. Hans-Achim Wagenknecht |
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Affiliation: | 1. Institute of Organic Chemistry, Karlsruhe Institute of Technology (KIT), Fritz-Haber-Weg 6, 76131 Karlsruhe, Germany;2. Institut Européen de Chimie et Biologie and ISM CNRS UMR5255, Université de Bordeaux, 2 Rue Robert Escarpit, 33607 Pessac, France |
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Abstract: | Sydnones are highly stable mesoionic 1,3-dipoles that react with cyclooctynes through strain-promoted sydnone-alkyne cycloaddition (SPSAC). Although sydnones have been shown to be valuable bioorthogonal chemical reporters for the labeling of proteins and complex glycans, nucleic acids have not yet been tagged by SPSAC. Evaluation of SPSAC kinetics with model substrates showed fast reactions with cyclooctyne probes (up to k=0.59 M−1 s−1), and two different sydnones were effectively incorporated into both 2’-deoxyuridines at position 5, and 7-deaza-2’-deoxyadenosines at position 7. These modified nucleosides were synthetically incorporated into single-stranded DNAs, which were successfully postsynthetically labeled with cyclooctyne probes both in vitro and in cells. These results show that sydnones are versatile bioorthogonal tags and have the premise to become essential tools for tracking DNA and potentially RNA in living cells. |
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Keywords: | cell biology cyclooctyne fluorescence oligonucleotide sydnone strain |
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