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Fluorescent Polymer-AS1411-Aptamer Probe for dSTORM Super-Resolution Imaging of Endogenous Nucleolin

Abstract : Nucleolin is a multifunctional protein involved in essential biological processes. To precisely localize it and unravel its different roles in cells, fluorescence imaging is a powerful tool, especially super-resolution techniques. Here, we developed polymer-aptamer probes, both small and bright, adapted to dSTORM nanoscopy. Well-defined fluorescent polymer chains bearing fluorophores (AlexaFluor647) and a reactive end-group were prepared via RAFT polymerization. The reactive end-group was then used for the oriented conjugation with AS1411, a DNA aptamer that recognizes nucleolin with high affinity. Conjugation via strained alkyne/azide click chemistry (SPAAC) between dibenzylcyclooctyne (DBCO)-ended fluorescent polymer chains and 3'-azidofunctionalized nucleic acids proved to be the most efficient approach. In vitro and in cellulo evaluations demonstrated that selective recognition for nucleolin was retained. Their brightness and small size make these polymer-aptamer probes an appealing alternative to immunofluorescence, especially for super-resolution (10-20 nm) nanoscopy. dSTORM imaging demonstrated the ability of our fluorescent polymer-aptamer probe to provide selective and superresolved detection of cell surface nucleolin.
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Contributor : Arnaud Favier Connect in order to contact the contributor
Submitted on : Tuesday, August 30, 2022 - 12:31:57 PM
Last modification on : Tuesday, September 27, 2022 - 4:20:45 AM


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Laura Fabre, Corentin Rousset, Karine Monier, Fernande da Cruz-Boisson, Philippe Bouvet, et al.. Fluorescent Polymer-AS1411-Aptamer Probe for dSTORM Super-Resolution Imaging of Endogenous Nucleolin. Biomacromolecules, American Chemical Society, 2022, 23 (6), pp.2302-2314. ⟨10.1021/acs.biomac.1c01706⟩. ⟨hal-03764508⟩



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