{"doi":"10.1101/2022.12.13.520107","title":"Next-generation plasmids for transgenesis in zebrafish and beyond","abstract":"Abstract Transgenesis is an essential technique for any genetic model. Tol2-based transgenesis paired with Gateway-compatible vector collections has transformed zebrafish transgenesis with an accessible, modular system. Here, we established several next-generation transgenesis tools for zebrafish and other species to expand and enhance transgenic applications. To facilitate gene-regulatory element testing, we generated Gateway middle entry vectors harboring the small mouse beta-globin minimal promoter coupled to several fluorophores, CreERT2, and Gal4. To extend the color spectrum for transgenic applications, we established middle entry vectors encoding the bright, blue-fluorescent protein mCerulean and mApple as an alternative red fluorophore. We present a series of p2A peptide-based 3’ vectors with different fluorophores and subcellular localizations to co-label cells expressing proteins of interest. Lastly, we established Tol2 destination vectors carrying the zebrafish exorh promoter driving different fluorophores as a pineal gland-specific transgenesis marker active prior to hatching and through adulthood. e xorh -based reporters and transgenesis markers also drive specific pineal gland expression in the eye-less cavefish ( Astyanax ). Together, our vectors provide versatile reagents for transgenesis applications in zebrafish, cavefish, and other models.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2022,"id":302364,"datarank":0.16479184330021646,"base_score":1.0986122886681096,"endowment":1.0986122886681096,"self_citation_contribution":0.16479184330021646,"citation_network_contribution":0.0,"self_endowment_contribution":0.16479184330021646,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":2,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9492,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":994360,"name":"Hannah R. Moran","orcid":"0000-0001-8056-5762","position":1,"is_corresponding":false},{"id":994361,"name":"Brooke F. Murray","orcid":"0000-0002-1351-3572","position":2,"is_corresponding":false},{"id":994713,"name":"Aaron Scoresby","orcid":null,"position":3,"is_corresponding":false},{"id":994714,"name":"John R. Klem","orcid":null,"position":4,"is_corresponding":false},{"id":994715,"name":"Rachel L. Eckert","orcid":null,"position":5,"is_corresponding":false},{"id":994716,"name":"Elizabeth Lepovsky","orcid":null,"position":6,"is_corresponding":false},{"id":585298,"name":"Sylvain Bertho","orcid":"0000-0002-1219-7352","position":7,"is_corresponding":false},{"id":994362,"name":"Susan Nieuwenhuize","orcid":"0009-0005-6979-8836","position":8,"is_corresponding":false},{"id":347201,"name":"Sibylle Bürger","orcid":"0009-0001-8861-0959","position":9,"is_corresponding":false},{"id":994363,"name":"Gianluca D‘Agati","orcid":"0000-0002-1021-0015","position":10,"is_corresponding":false},{"id":994364,"name":"Charles Betz","orcid":"0000-0002-9837-6916","position":11,"is_corresponding":false},{"id":994717,"name":"Ann‐Christin Puller","orcid":null,"position":12,"is_corresponding":false},{"id":994365,"name":"Anastasia Felker","orcid":"0000-0002-3734-2026","position":13,"is_corresponding":false},{"id":994366,"name":"Karolína Ditrychová","orcid":"0000-0002-2209-2941","position":14,"is_corresponding":false},{"id":994367,"name":"Seraina Bötschi","orcid":"0000-0002-2721-133X","position":15,"is_corresponding":false},{"id":351905,"name":"Markus Affolter","orcid":"0000-0002-5171-0016","position":16,"is_corresponding":false},{"id":560930,"name":"Nicolas Rohner","orcid":"0000-0003-3248-2772","position":17,"is_corresponding":false},{"id":705205,"name":"C. Ben Lovely","orcid":"0000-0002-7838-5823","position":18,"is_corresponding":false},{"id":254324,"name":"Kristen M. Kwan","orcid":"0000-0003-0052-275X","position":19,"is_corresponding":false},{"id":994368,"name":"Alexa Burger","orcid":"0000-0001-7137-3910","position":20,"is_corresponding":false},{"id":652950,"name":"Christian Mosimann","orcid":"0000-0002-0749-2576","position":21,"is_corresponding":false},{"id":994359,"name":"Cassie L. Kemmler","orcid":"0000-0002-4516-029X","position":0,"is_corresponding":true}],"reference_count":135,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T00:32:16.279991Z","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":[]}