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

October 2018. Accurate labelling of endogenous proteins for advanced light microscopy of living cells remains challenging. Nanobodies have been widely used for antigen labeling, visualization of subcellular protein localization and interactions. Nanobodies are very small antibodies that have only a single, monomeric domain.

To facilitate wider application, scientists at Goethe University Frankfurt developed a scalable and high-throughput strategy to simultaneously target multiple endogenous proteins in living cells with micro- to nanometer resolution. For intracellular protein labelling, they advanced nanobodies by site-specific and stoichiometric attachment of bright organic fluorophores.

Their fast and fine-tuned intracellular transfer by microfluidic cell squeezing enables high-throughput delivery with less than 10 percent cell mortality. The strategy allows for the dual-colour imaging of distinct endogenous cellular structures, and allows 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.

 

Contact:
Robert Tampé and Ralph Wieneke, Institute of Biochemistry, Riedberg Campus, Goethe University, Frankfurt/Main, Germany, tampe@em.uni-frankfurt.de

 

Publication:
Klein A, Hank S, Raulf A, Joest EF, Tissen F, Heilemann M, Wieneke R*, Tampé R* (2018) Live-cell labeling of endogenous proteins with nanometer precision by transduced nanobodies. Chemical Science 9: 7835-7842. http://dx.doi.org/10.1039/C8SC02910E

 

 

 

Cluster of Excellence Macromolecular Complexes, Frankfurt am Main, Germany