Advanced Fluorescent Polymer Probes for the Site-Specific Labeling of Proteins in Live Cells Using the HaloTag Technology

使用 HaloTag 技术对活细胞中的蛋白质进行位点特异性标记的先进荧光聚合物探针

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作者:Thomas Berki, Anush Bakunts, Damien Duret, Laura Fabre, Catherine Ladavière, Andrea Orsi, Marie-Thérèse Charreyre, Andrea Raimondi, Eelco van Anken, Arnaud Favier

Abstract

We report the site-specific and covalent bioconjugation of fluorescent polymer chains to proteins in live cells using the HaloTag technology. Polymer chains bearing a Halo-ligand precisely located at their α-chain-end were synthesized in a controlled manner owing to the RAFT polymerization process. They were labeled in lateral position by several organic fluorophores such as AlexaFluor 647. The resulting Halo-ligand polymer probe was finally shown to selectively recognize and label HaloTag proteins present at the membrane of live cells using confocal fluorescence microscopy. Such a polymer bioconjugation approach holds great promises for various applications ranging from cell imaging to cell surface functionalization.

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