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




TekijätKatrin Daniel, Jaroslav Icha, Cindy Horenburg, Doris Müller, Caren Norden, Jörg Mansfeld

KustantajaNATURE PUBLISHING GROUP

Julkaisuvuosi2018

Lehti:Nature Communications

Tietokannassa oleva lehden nimiNATURE COMMUNICATIONS

Lehden akronyymiNAT COMMUN

Artikkelin numero3297

Vuosikerta9

Aloitussivu1

Lopetussivu13

Sivujen määrä13

ISSN2041-1723

DOIhttps://doi.org/10.1038/s41467-018-05855-5

Rinnakkaistallenteen osoitehttps://research.utu.fi/converis/portal/detail/Publication/35818381


Tiivistelmä
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.

Ladattava julkaisu

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Last updated on 2024-26-11 at 10:34