核仁素
适体
荧光
生物物理学
聚合物
纳米技术
化学
材料科学
生物化学
生物
分子生物学
有机化学
物理
量子力学
细胞质
核仁
作者
Laura Fabre,Corentin Rousset,Karine Monier,Fernande Da Cruz-Boisson,Philippe Bouvet,Marie‐Thérèse Charreyre,Thierry Delair,Étienne Fleury,Arnaud Favier
出处
期刊:Biomacromolecules
[American Chemical Society]
日期:2022-05-12
卷期号:23 (6): 2302-2314
被引量:5
标识
DOI:10.1021/acs.biomac.1c01706
摘要
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 direct stochastic optical reconstruction microscopy (dSTORM). 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 strain-promoted alkyne/azide click chemistry (SPAAC) between dibenzylcyclooctyne-ended fluorescent polymer chains and 3′-azido-functionalized 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 super-resolved detection of cell surface nucleolin.
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