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

Katrin Daniel, Jaroslav Icha, Cindy Horenburg, Doris Müller, Caren Norden () and Jörg Mansfeld ()
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Katrin Daniel: Technische Universität Dresden
Jaroslav Icha: Max Planck Institute of Molecular Cell Biology and Genetics
Cindy Horenburg: Technische Universität Dresden
Doris Müller: Technische Universität Dresden
Caren Norden: Max Planck Institute of Molecular Cell Biology and Genetics
Jörg Mansfeld: Technische Universität Dresden

Nature Communications, 2018, vol. 9, issue 1, 1-13

Abstract: 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.

Date: 2018
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DOI: 10.1038/s41467-018-05855-5

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