期刊
NATURE COMMUNICATIONS
卷 9, 期 -, 页码 -出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/s41467-018-05855-5
关键词
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资金
- German Research Foundation (DFG) [MA 5831/1-1, SFB 655]
- European Research Council (ERC) under the European Union's Horizon 2020 research and innovation program [680042]
- Maria-Reiche-Program of the TU Dresden
- Human Frontier Science Program [CDA-00007/2011]
- Fish and Light Microscopy Facilities of the Biotechnology Center of the TU Dresden
- Max Planck Institute of Molecular Cell Biology and Genetics
- Open Access Publication Funds of the SLUB/TU Dresden
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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