{"doi":"10.1101/2020.05.12.090217","title":"Expanding the <i>Caenorhabditis elegans</i> auxin-inducible degron system toolkit with internal expression and degradation controls and improved modular constructs for CRISPR/Cas9-mediated genome editing","abstract":"Abstract 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, hypodermis, seam cell, anchor cell) and pan-somatic TIR1- expressing strains carrying an equimolar co-expressed blue fluorescent reporter to enable use of both red and green channels in experiments. We have also constructed a set of plasmids to generate fluorescent protein::AID fusions through CRISPR/Cas9-mediated genome editing. These templates can be produced through frequently used cloning systems (Gibson assembly or SapTrap) or through ribonucleoprotein complex-mediated insertion of PCR-derived, linear repair templates. We have generated a set of sgRNA plasmids carrying modifications shown to boost editing efficiency, targeting standardized transgene insertion sites on chromosomes I and II. Together these reagents should complement existing TIR1 strains and facilitate rapid and high-throughput fluorescent protein::AID* tagging of factors of interest. This battery of new TIR1-expressing strains and modular, efficient cloning vectors serves as a platform for facile assembly of CRISPR/Cas9 repair templates for conditional protein depletion.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2020,"id":120908,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":8,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9389,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":475287,"name":"Tam Duong","orcid":"0000-0002-0754-0355","position":1,"is_corresponding":false},{"id":559267,"name":"Max T. Levenson","orcid":"0000-0002-1470-3968","position":2,"is_corresponding":false},{"id":308271,"name":"Michael A. Q. Martinez","orcid":"0000-0003-1178-7139","position":3,"is_corresponding":false},{"id":271687,"name":"Jonathan D. Hibshman","orcid":"0000-0001-7324-9382","position":4,"is_corresponding":false},{"id":492346,"name":"Hannah N. Saeger","orcid":"0000-0002-3055-5547","position":5,"is_corresponding":false},{"id":559959,"name":"Ryan Doonan","orcid":null,"position":6,"is_corresponding":false},{"id":308272,"name":"Nicholas J. Palmisano","orcid":"0000-0002-7992-4462","position":7,"is_corresponding":false},{"id":492345,"name":"Raquel Martínez-Méndez","orcid":"0000-0003-4219-8003","position":8,"is_corresponding":false},{"id":559268,"name":"Brittany Davidson","orcid":"0000-0003-3538-2822","position":9,"is_corresponding":false},{"id":308278,"name":"Wan Zhang","orcid":"0000-0003-1896-8186","position":10,"is_corresponding":false},{"id":492342,"name":"James Matthew Ragle","orcid":"0000-0002-6626-2615","position":11,"is_corresponding":false},{"id":308274,"name":"Taylor N. Medwig-Kinney","orcid":"0000-0001-7989-3291","position":12,"is_corresponding":false},{"id":559269,"name":"Sydney S. Sirota","orcid":"0000-0001-9990-3266","position":13,"is_corresponding":false},{"id":271688,"name":"Bob Goldstein","orcid":"0000-0001-6961-675X","position":14,"is_corresponding":false},{"id":308284,"name":"David Q. Matus","orcid":"0000-0002-1570-5025","position":15,"is_corresponding":false},{"id":361111,"name":"Daniel J. Dickinson","orcid":"0000-0003-2651-2584","position":16,"is_corresponding":false},{"id":475290,"name":"David J. Reiner","orcid":"0000-0002-0344-7161","position":17,"is_corresponding":false},{"id":492350,"name":"Jordan D. Ward","orcid":"0000-0001-9870-8936","position":18,"is_corresponding":false},{"id":492347,"name":"Guinevere Ashley","orcid":"0000-0002-3983-5553","position":0,"is_corresponding":true}],"reference_count":73,"raw_metadata":null,"created_at":"2026-07-18T23:14:38.147936Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}