A1 Refereed original research article in a scientific journal
Conditional control of fluorescent protein degradation by an auxin-dependent nanobody
Authors: Katrin Daniel, Jaroslav Icha, Cindy Horenburg, Doris Müller, Caren Norden, Jörg Mansfeld
Publisher: NATURE PUBLISHING GROUP
Publication year: 2018
Journal:: Nature Communications
Journal name in source: NATURE COMMUNICATIONS
Journal acronym: NAT COMMUN
Article number: 3297
Volume: 9
First page : 1
Last page: 13
Number of pages: 13
ISSN: 2041-1723
DOI: https://doi.org/10.1038/s41467-018-05855-5
Self-archived copy’s web address: https://research.utu.fi/converis/portal/detail/Publication/35818381
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.
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