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

Daniel, K., Icha, J., Horenburg, C., Müller, D., Norden, C., & Mansfeld, J. (2018). Conditional control of fluorescent protein degradation by an auxin-dependent nanobody. Nature communications, 9(1): 3297. doi:10.1038/s41467-018-05855-5.

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 Creators:
Daniel, Katrin, Author
Icha, Jaroslav1, Author           
Horenburg, Cindy, Author
Müller, Doris, Author
Norden, Caren1, Author           
Mansfeld, Jorg, Author
Affiliations:
1Max Planck Institute for Molecular Cell Biology and Genetics, Max Planck Society, ou_2340692              

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 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.

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 Dates: 2018-08-17
 Publication Status: Issued
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 Identifiers: DOI: 10.1038/s41467-018-05855-5
Other: cbg-7197
PMID: 30120238
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Title: Nature communications
  Other : Nat Commun
Source Genre: Journal
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Pages: - Volume / Issue: 9 (1) Sequence Number: 3297 Start / End Page: - Identifier: -