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

Author

Listed:
  • Katrin Daniel

    (Technische Universität Dresden)

  • Jaroslav Icha

    (Max Planck Institute of Molecular Cell Biology and Genetics
    University of Turku)

  • Cindy Horenburg

    (Technische Universität Dresden)

  • Doris Müller

    (Technische Universität Dresden)

  • Caren Norden

    (Max Planck Institute of Molecular Cell Biology and Genetics)

  • Jörg Mansfeld

    (Technische Universität Dresden)

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.

Suggested Citation

  • Katrin Daniel & Jaroslav Icha & Cindy Horenburg & Doris Müller & Caren Norden & Jörg Mansfeld, 2018. "Conditional control of fluorescent protein degradation by an auxin-dependent nanobody," Nature Communications, Nature, vol. 9(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:9:y:2018:i:1:d:10.1038_s41467-018-05855-5
    DOI: 10.1038/s41467-018-05855-5
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