期刊
NATURE CHEMISTRY
卷 13, 期 1, 页码 15-+出版社
NATURE PORTFOLIO
DOI: 10.1038/s41557-020-00584-z
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资金
- Deutsche Forschungsgemeinschaft [SPP 1623, SFB 765]
DNA nanotechnology offers opportunities for manipulating and imaging proteins on cell surfaces by creating nucleic acid-protein junctions using PNA tags. This method allows for quick assembly of different fluorescent dyes, increased brightness, reversible labeling, and is applicable to various cell and protein systems.
DNA nanotechnology is an emerging field that promises fascinating opportunities for the manipulation and imaging of proteins on a cell surface. The key to progress is the ability to create a nucleic acid-protein junction in the context of living cells. Here we report a covalent labelling reaction that installs a biostable peptide nucleic acid (PNA) tag. The reaction proceeds within minutes and is specific for proteins carrying a 2 kDa coiled-coil peptide tag. Once installed, the PNA label serves as a generic landing platform that enables the recruitment of fluorescent dyes via nucleic acid hybridization. We demonstrate the versatility of this approach by recruiting different fluorophores, assembling multiple fluorophores for increased brightness and achieving reversible labelling by way of toehold-mediated strand displacement. Additionally, we show that labelling can be carried out using two different coiled-coil systems, with epidermal growth factor receptor and endothelin receptor type B, on both HEK293 and CHO cells. Finally, we apply the method to monitor internalization of epidermal growth factor receptor on CHO cells.
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