An expanded auxin-inducible degron toolkit for Caenorhabditis elegans. Academic Article uri icon


  • The auxin-inducible degron (AID) system has emerged as a powerful tool to conditionally deplete proteins in a range of organisms and cell types. Here, we describe a toolkit to augment the use of the AID system in Caenorhabditis elegans. We have generated a set of single-copy, tissue-specific (germline, intestine, neuron, muscle, pharynx, hypodermis, seam cell, anchor cell) and pan-somatic TIR1-expressing strains carrying a co-expressed blue fluorescent reporter to enable use of both red and green channels in experiments. These transgenes are inserted into commonly used, well-characterized genetic loci. We confirmed that our TIR1-expressing strains produce the expected depletion phenotype for several nuclear and cytoplasmic AID-tagged endogenous substrates. We have also constructed a set of plasmids for constructing repair templates to generate fluorescent protein::AID fusions through CRISPR/Cas9-mediated genome editing. These plasmids are compatible with commonly used genome editing approaches in the C. elegans community (Gibson or SapTrap assembly of plasmid repair templates or PCR-derived linear repair templates). Together these reagents will complement existing TIR1 strains and facilitate rapid and high-throughput fluorescent protein::AID tagging of genes. This battery of new TIR1-expressing strains and modular, efficient cloning vectors serves as a platform for straightforward assembly of CRISPR/Cas9 repair templates for conditional protein depletion.

published proceedings

  • Genetics

altmetric score

  • 33.076

author list (cited authors)

  • Ashley, G. E., Duong, T., Levenson, M. T., Martinez, M., Johnson, L. C., Hibshman, J. D., ... Ward, J. D.

citation count

  • 36

complete list of authors

  • Ashley, Guinevere E||Duong, Tam||Levenson, Max T||Martinez, Michael AQ||Johnson, Londen C||Hibshman, Jonathan D||Saeger, Hannah N||Palmisano, Nicholas J||Doonan, Ryan||Martinez-Mendez, Raquel||Davidson, Brittany R||Zhang, Wan||Ragle, James Matthew||Medwig-Kinney, Taylor N||Sirota, Sydney S||Goldstein, Bob||Matus, David Q||Dickinson, Daniel J||Reiner, David J||Ward, Jordan D

editor list (cited editors)

  • Greenstein, D.

publication date

  • March 2021