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Nat Commun. 2018 Aug 17;9(1):3297. doi: 10.1038/s41467-018-05855-5.

Conditional control of fluorescent protein degradation by an auxin-dependent nanobody.

Author information

1
Cell Cycle, Biotechnology Center, Technische Universität Dresden, Tatzberg 47-49, 01307, Dresden, Germany.
2
Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstraße 108, 01307, Dresden, Germany.
3
Turku Centre for Biotechnology, University of Turku, Tykistokatu 6, 20520, Turku, Finland.
4
Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstraße 108, 01307, Dresden, Germany. norden@mpi-cbg.de.
5
Cell Cycle, Biotechnology Center, Technische Universität Dresden, Tatzberg 47-49, 01307, Dresden, Germany. joerg.mansfeld@tu-dresden.de.

Abstract

The conditional and reversible depletion of proteins by auxin-mediated degradation is a powerful tool to investigate protein functions in cells and whole organisms. However, its wider applications require fusing the auxin-inducible degron (AID) to individual target proteins. Thus, establishing the auxin system for multiple proteins can be challenging. Another approach for directed protein degradation are anti-GFP nanobodies, which can be applied to GFP stock collections that are readily available in different experimental models. Here, we combine the advantages of auxin and nanobody-based degradation technologies creating an AID-nanobody to degrade GFP-tagged proteins at different cellular structures in a conditional and reversible manner in human cells. We demonstrate efficient and reversible inactivation of the anaphase promoting complex/cyclosome (APC/C) and thus provide new means to study the functions of this essential ubiquitin E3 ligase. Further, we establish auxin degradation in a vertebrate model organism by employing AID-nanobodies in zebrafish.

PMID:
30120238
PMCID:
PMC6098157
DOI:
10.1038/s41467-018-05855-5
[Indexed for MEDLINE]
Free PMC Article

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