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

  • 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.

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Author:Alina Klein, Susanne Hank, Anika Raulf, Eike F. Joest, Felicitas Tissen, Mike HeilemannORCiDGND, Ralph WienekeORCiDGND, Robert TampéORCiDGND
Pubmed Id:
Parent Title (English):Chemical science
Place of publication:Cambridge
Document Type:Article
Year of Completion:2018
Date of first Publication:2018/08/20
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2018/12/20
Issue:40, Art. 7835
Page Number:8
First Page:1
Last Page:8
This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence.
Institutes:Biochemie, Chemie und Pharmazie / Biochemie und Chemie
Wissenschaftliche Zentren und koordinierte Programme / Sonderforschungsbereiche / Forschungskollegs
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 54 Chemie / 540 Chemie und zugeordnete Wissenschaften
Licence (German):License LogoCreative Commons - Namensnennung-Nicht kommerziell 3.0