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bioRxiv · 10.1101/298257

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

Daniel, K.·Icha, J.·Horenburg, C.·Muller, D.·Norden, C.·Mansfeld, J.
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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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Daniel, K., Icha, J., Horenburg, C., Muller, D., Norden, C., Mansfeld, J.. 2018-04-09. Conditional control of fluorescent protein degradation by an auxin-dependent nanobody. https://doi.org/10.1101/298257

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