Issue 16, 2021

Light-guided intrabodies for on-demand in situ target recognition in human cells

Abstract

Due to their high stability and specificity in living cells, fluorescently labeled nanobodies are perfect probes for visualizing intracellular targets at an endogenous level. However, intrabodies bind unrestrainedly and hence may interfere with the target protein function. Here, we report a strategy to prevent premature binding through the development of photo-conditional intrabodies. Using genetic code expansion, we introduce photocaged amino acids within the nanobody-binding interface, which, after photo-activation, show instantaneous binding of target proteins with high spatiotemporal precision inside living cells. Due to the highly stable binding, light-guided intrabodies offer a versatile platform for downstream imaging and regulation of target proteins.

Graphical abstract: Light-guided intrabodies for on-demand in situ target recognition in human cells

Supplementary files

Article information

Article type
Edge Article
Submitted
07 mar 2021
Accepted
22 mar 2021
First published
08 apr 2021
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY-NC license

Chem. Sci., 2021,12, 5787-5795

Light-guided intrabodies for on-demand in situ target recognition in human cells

E. F. Joest, C. Winter, J. S. Wesalo, A. Deiters and R. Tampé, Chem. Sci., 2021, 12, 5787 DOI: 10.1039/D1SC01331A

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