- 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(s): Greenstein, D
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.