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Development. 2015 Dec 15;142(24):4374-84. doi: 10.1242/dev.129635. Epub 2015 Nov 9.

The auxin-inducible degradation (AID) system enables versatile conditional protein depletion in C. elegans.

Author information

1
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720-3220, USA Howard Hughes Medical Institute, 4000 Jones Bridge Road, Chevy Chase, MD 20815, USA Life Sciences Division, Department of Genome Dynamics, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA California Institute for Quantitative Biosciences, Berkeley, CA 94720, USA.
2
Department of Cellular and Molecular Pharmacology, University of California, San Francisco, CA 94158, USA.
3
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720-3220, USA.
4
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720-3220, USA Howard Hughes Medical Institute, 4000 Jones Bridge Road, Chevy Chase, MD 20815, USA Life Sciences Division, Department of Genome Dynamics, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA California Institute for Quantitative Biosciences, Berkeley, CA 94720, USA afdernburg@berkeley.edu.

Abstract

Experimental manipulation of protein abundance in living cells or organisms is an essential strategy for investigation of biological regulatory mechanisms. Whereas powerful techniques for protein expression have been developed in Caenorhabditis elegans, existing tools for conditional disruption of protein function are far more limited. To address this, we have adapted the auxin-inducible degradation (AID) system discovered in plants to enable conditional protein depletion in C. elegans. We report that expression of a modified Arabidopsis TIR1 F-box protein mediates robust auxin-dependent depletion of degron-tagged targets. We document the effectiveness of this system for depletion of nuclear and cytoplasmic proteins in diverse somatic and germline tissues throughout development. Target proteins were depleted in as little as 20-30 min, and their expression could be re-established upon auxin removal. We have engineered strains expressing TIR1 under the control of various promoter and 3' UTR sequences to drive tissue-specific or temporally regulated expression. The degron tag can be efficiently introduced by CRISPR/Cas9-based genome editing. We have harnessed this system to explore the roles of dynamically expressed nuclear hormone receptors in molting, and to analyze meiosis-specific roles for proteins required for germ line proliferation. Together, our results demonstrate that the AID system provides a powerful new tool for spatiotemporal regulation and analysis of protein function in a metazoan model organism.

KEYWORDS:

Auxin; Auxin-inducible degradation; C. elegans; Degron; Genetic tool; Tissue-specific depletion

PMID:
26552885
PMCID:
PMC4689222
DOI:
10.1242/dev.129635
[Indexed for MEDLINE]
Free PMC Article

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