{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T03:40:32Z","timestamp":1792813232100,"version":"4.5.0"},"reference-count":28,"publisher":"The Company of Biologists","issue":"10","license":[{"start":{"date-parts":[[2020,9,3]],"date-time":"2020-09-03T00:00:00Z","timestamp":1599091200000},"content-version":"vor","delay-in-days":703,"URL":"http:\/\/creativecommons.org\/licenses\/by\/3.0"}],"funder":[{"DOI":"10.13039\/501100002996","name":"Dutch Heart Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002996","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100017732","name":"E-Rare","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100017732","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2018,10,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n               <jats:p>The zebrafish (Danio rerio) has become a popular vertebrate model organism to study organ formation and function due to its optical clarity and rapid embryonic development. The use of genetically modified zebrafish has also allowed identification of new putative therapeutic drugs. So far, most studies have relied on broad overexpression of transgenes harboring patient-derived mutations or loss-of-function mutants, which incompletely model the human disease allele in terms of expression levels or cell-type specificity of the endogenous gene of interest. Most human genetically inherited conditions are caused by alleles carrying single nucleotide changes resulting in altered gene function. Introduction of such point mutations in the zebrafish genome would be a prerequisite to recapitulate human disease but remains challenging to this day. We present an effective approach to introduce small nucleotide changes in the zebrafish genome. We generated four different knock-in lines carrying distinct human cardiovascular-disorder-causing missense mutations in their zebrafish orthologous genes by combining CRISPR\/Cas9 with a short template oligonucleotide. Three of these lines carry gain-of-function mutations in genes encoding the pore-forming (Kir6.1, KCNJ8) and regulatory (SUR2, ABCC9) subunits of an ATP-sensitive potassium channel (KATP) linked to Cant\u00fa syndrome (CS). Our heterozygous zebrafish knock-in lines display significantly enlarged ventricles with enhanced cardiac output and contractile function, and distinct cerebral vasodilation, demonstrating the causality of the introduced mutations for CS. These results demonstrate that introducing patient alleles in their zebrafish orthologs promises a broad application for modeling human genetic diseases, paving the way for new therapeutic strategies using this model organism.<\/jats:p>","DOI":"10.1242\/dmm.035469","type":"journal-article","created":{"date-parts":[[2018,10,18]],"date-time":"2018-10-18T13:16:22Z","timestamp":1539868582000},"source":"Crossref","is-referenced-by-count":88,"title":["Effective CRISPR\/Cas9-based nucleotide editing in zebrafish to model human genetic cardiovascular disorders"],"prefix":"10.1242","volume":"11","author":[{"given":"Federico","family":"Tessadori","sequence":"first","affiliation":[{"name":"Hubrecht Institute-KNAW and UMC Utrecht 1 , 3584 CT Utrecht , the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helen I.","family":"Roessler","sequence":"additional","affiliation":[{"name":"Center for Molecular Medicine, University Medical Center Utrecht 2 Department of Genetics , , 3584 CX Utrecht , the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sanne M. C.","family":"Savelberg","sequence":"additional","affiliation":[{"name":"Center for Molecular Medicine, University Medical Center Utrecht 2 Department of Genetics , , 3584 CX Utrecht , the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sonja","family":"Chocron","sequence":"additional","affiliation":[{"name":"Hubrecht Institute-KNAW and UMC Utrecht 1 , 3584 CT Utrecht , the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah M.","family":"Kamel","sequence":"additional","affiliation":[{"name":"Hubrecht Institute-KNAW and UMC Utrecht 1 , 3584 CT Utrecht , the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karen J.","family":"Duran","sequence":"additional","affiliation":[{"name":"Center for Molecular Medicine, University Medical Center Utrecht 2 Department of Genetics , , 3584 CX Utrecht , the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mieke M.","family":"van Haelst","sequence":"additional","affiliation":[{"name":"Center for Molecular Medicine, University Medical Center Utrecht 2 Department of Genetics , , 3584 CX Utrecht , the Netherlands"},{"name":"Amsterdam Medical Center, University of Amsterdam 3 Department of Clinical Genetics , , 1105 AZ Amsterdam , the Netherlands"},{"name":"Free University Medical Center 4 Department of Clinical Genetics , , 1018 HV Amsterdam , the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3033-0329","authenticated-orcid":false,"given":"Gijs","family":"van Haaften","sequence":"additional","affiliation":[{"name":"Center for Molecular Medicine, University Medical Center Utrecht 2 Department of Genetics , , 3584 CX Utrecht , the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9418-0422","authenticated-orcid":false,"given":"Jeroen","family":"Bakkers","sequence":"additional","affiliation":[{"name":"Hubrecht Institute-KNAW and UMC Utrecht 1 , 3584 CT Utrecht , the Netherlands"},{"name":"University Medical Center Utrecht 5 Department of Medical Physiology, Division of Heart and Lungs , , 3584 CX Utrecht , the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"237","published-online":{"date-parts":[[2018,10,18]]},"reference":[{"key":"2024061302055136800_DMM035469C1","doi-asserted-by":"publisher","first-page":"e0150188","DOI":"10.1371\/journal.pone.0150188","article-title":"Homology directed knockin of point mutations in the zebrafish tardbp and fus genes in ALS using the CRISPR\/Cas9 system","volume":"11","author":"Armstrong","year":"2016","journal-title":"PLoS ONE"},{"key":"2024061302055136800_DMM035469C2","doi-asserted-by":"publisher","first-page":"179","DOI":"10.1016\/j.cmet.2010.02.004","article-title":"New uses for old drugs: neonatal diabetes and sulphonylureas","volume":"11","author":"Ashcroft","year":"2010","journal-title":"Cell Metab."},{"key":"2024061302055136800_DMM035469C3","doi-asserted-by":"publisher","first-page":"114","DOI":"10.1038\/nature11537","article-title":"In vivo genome editing using a high-efficiency TALEN system","volume":"491","author":"Bedell","year":"2012","journal-title":"Nature"},{"key":"2024061302055136800_DMM035469C4","doi-asserted-by":"publisher","first-page":"678","DOI":"10.1016\/j.ejmg.2013.09.009","article-title":"Mutation of KCNJ8 in a patient with Cantu syndrome with unique vascular abnormalities-support for the role of K(ATP) channels in this condition","volume":"56","author":"Brownstein","year":"2013","journal-title":"Eur. 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