{"doi":"10.1101/2022.09.06.506849","title":"Generation of floxed alleles for cell-specific knockout in zebrafish","abstract":"SUMMARY A benefit of the zebrafish as a model is its amenability to genetic approaches. However, a lack of conditional deletion alleles makes it a challenge to perform stage- or cell-specific knockout. Here, we initially applied an existing protocol to establish a floxed allele for gata2a and encountered several issues, including off-target integration and incomplete knock-in. To address these problems, we developed a protocol incorporating simultaneous co-targeting with Cas12a to insert loxP sites in cis , together with transgenic counter-screening to identify off-target insertions. We applied this protocol to establish a floxed allele of foxc1a in a single generation. We demonstrate the utility of our floxed alleles by verifying Cre-dependent deletion, which yielded expected phenotypes in each case. Finally, we used the floxed gata2a allele to demonstrate an endothelial autonomous requirement in lymphatic valve development. Together, our results provide a framework for straightforward generation and application of floxed alleles in zebrafish.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2022,"id":304419,"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":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9464,"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":997813,"name":"Takayuki Nozaki","orcid":null,"position":1,"is_corresponding":false},{"id":997814,"name":"Benjamin J. Toles","orcid":null,"position":2,"is_corresponding":false},{"id":694420,"name":"Amy Kolb","orcid":null,"position":3,"is_corresponding":false},{"id":588036,"name":"Kevin Luk","orcid":"0000-0002-9727-1960","position":4,"is_corresponding":false},{"id":997276,"name":"Sumio Isogai","orcid":"0000-0002-9290-4650","position":5,"is_corresponding":false},{"id":997815,"name":"Kinji Ishida","orcid":null,"position":6,"is_corresponding":false},{"id":997816,"name":"Tomohito Hanasaka","orcid":null,"position":7,"is_corresponding":false},{"id":997277,"name":"Michael Parsons","orcid":"0000-0002-4046-1659","position":8,"is_corresponding":false},{"id":588037,"name":"Scot A. Wolfe","orcid":"0000-0002-7042-201X","position":9,"is_corresponding":false},{"id":304178,"name":"Nathan D. Lawson","orcid":"0000-0001-7788-9619","position":10,"is_corresponding":false},{"id":649394,"name":"Masahiro Shin","orcid":"0000-0002-0231-894X","position":0,"is_corresponding":true}],"reference_count":50,"raw_metadata":null,"created_at":"2026-07-19T00:32:32.651796Z","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":[]}