title: The auxin-inducible degradation (AID) system enables versatile conditional protein depletion in C. elegans.
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.
authors.affiliation: 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.
authors.forename: Liangyu
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, CA 94158, USA.
authors.forename: Jordan D
authors.initials: JD
authors.lastname: Ward
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720-3220, USA.
authors.forename: Ze
authors.initials: Z
authors.lastname: Cheng
authors.affiliation: 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.
authors.forename: Abby F
authors.initials: AF
authors.lastname: Dernburg
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000378:metabolism; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism; D020964:Embryo Loss / Q000473:pathology; D005298:Fertility / Q000187:drug effects; D017353:Gene Deletion; D005854:Germ Cells / Q000187:drug effects / Q000378:metabolism; D007210:Indoleacetic Acids / Q000494:pharmacology*; D007814:Larva / Q000187:drug effects; D008540:Meiosis / Q000187:drug effects; D009687:Nuclear Proteins / Q000378:metabolism; D009928:Organ Specificity / Q000187:drug effects; D059748:Proteolysis / Q000187:drug effects*; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism
chemical_list: D029742:Caenorhabditis elegans Proteins; D007210:Indoleacetic Acids; D009687:Nuclear Proteins; D018160:Receptors, Cytoplasmic and Nuclear
keywords: Auxin; Auxin-inducible degradation; C. elegans; Degron; Genetic tool; Tissue-specific depletion