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Issue 40, 2018
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Live-cell labeling of endogenous proteins with nanometer precision by transduced nanobodies

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Abstract

Accurate labeling of endogenous proteins for advanced light microscopy in living cells remains challenging. Nanobodies have been widely used for antigen labeling, visualization of subcellular protein localization and interactions. To facilitate an expanded application, we present a scalable and high-throughput strategy to simultaneously target multiple endogenous proteins in living cells with micro- to nanometer resolution. For intracellular protein labeling, we advanced nanobodies by site-specific and stoichiometric attachment of bright organic fluorophores. Their fast and fine-tuned intracellular transfer by microfluidic cell squeezing enabled high-throughput delivery with less than 10% dead cells. This strategy allowed for the dual-color imaging of distinct endogenous cellular structures, and culminated in super-resolution imaging of native protein networks in genetically non-modified living cells. The simultaneous delivery of multiple engineered nanobodies does not only offer exciting prospects for multiplexed imaging of endogenous protein, but also holds potential for visualizing native cellular structures with unprecedented accuracy.

Graphical abstract: Live-cell labeling of endogenous proteins with nanometer precision by transduced nanobodies

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Publication details

The article was received on 02 Jul 2018, accepted on 20 Aug 2018 and first published on 20 Aug 2018


Article type: Edge Article
DOI: 10.1039/C8SC02910E
Citation: Chem. Sci., 2018,9, 7835-7842
  • Open access: Creative Commons BY-NC license
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    Live-cell labeling of endogenous proteins with nanometer precision by transduced nanobodies

    A. Klein, S. Hank, A. Raulf, E. F. Joest, F. Tissen, M. Heilemann, R. Wieneke and R. Tampé, Chem. Sci., 2018, 9, 7835
    DOI: 10.1039/C8SC02910E

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