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

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

Publisher: NATURE PUBLISHING GROUP

: 2018

: Nature Communications

: NATURE COMMUNICATIONS

: NAT COMMUN

: 3297

: 9

: 1

: 13

: 2041-1723

DOI: https://doi.org/10.1038/s41467-018-05855-5

: 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.

s41467-018-05855-5.pdf