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Live cell PNA labelling enables erasable fluorescence imaging of membrane proteins. | LitMetric

AI Article Synopsis

  • - DNA nanotechnology is advancing the manipulation and imaging of proteins on cell surfaces, focusing on creating specific nucleic acid-protein junctions in living cells.
  • - Researchers developed a rapid, covalent labeling method using a biostable peptide nucleic acid (PNA) tag that targets proteins with a specific coiled-coil peptide, allowing for the recruitment of fluorescent dyes.
  • - The technique was successfully demonstrated on various cells and proteins, enabling versatile applications such as enhanced brightness with multiple fluorophores and real-time monitoring of protein internalization, specifically the epidermal growth factor receptor.

Article Abstract

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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Source
http://dx.doi.org/10.1038/s41557-020-00584-zDOI Listing

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