An expanded auxin-inducible degron toolkit for Caenorhabditis elegans.
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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.
author list (cited authors)
Ashley, G. E., Duong, T., Levenson, M. T., Martinez, M., Johnson, L. C., Hibshman, J. D., ... Ward, J. D.
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)