{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T21:42:40Z","timestamp":1760391760747,"version":"build-2065373602"},"reference-count":26,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2020,8,5]],"date-time":"2020-08-05T00:00:00Z","timestamp":1596585600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000065","name":"National Institute of Neurological Disorders and Stroke","doi-asserted-by":"publisher","award":["R01NS082645"],"award-info":[{"award-number":["R01NS082645"]}],"id":[{"id":"10.13039\/100000065","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["anatomypubs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Developmental Dynamics"],"published-print":{"date-parts":[[2020,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>With the goal of labeling and manipulating the zebrafish hypothalamus, we sought to target a <jats:italic>green fluorescent protein (gfp)<\/jats:italic> transgene to the expression domains of <jats:italic>nkx2.4b<\/jats:italic>, a gene expressed during hypothalamic and thyroid development. We combined transcription activator\u2010like effector nucleases (TALENs)\u2010mediated mutagenesis with a targeting construct to enable insertion of a <jats:italic>gfp<\/jats:italic> transgene into the endogenous <jats:italic>nkx2.4b<\/jats:italic> genomic locus.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Injection of TALENs targeted to the first exon of <jats:italic>nkx2.4b<\/jats:italic> created a predicted null allele, and homozygous mutant embryos displayed loss of thyroid markers. From embryos injected with both TALENs and a targeting construct carrying a <jats:italic>gfp<\/jats:italic> transgene, we recovered a line in which GFP was expressed specifically in the hypothalamus and thyroid. Fish homozygous for this allele lacked exon 1 of <jats:italic>nkx2.4b<\/jats:italic> and exhibited hypothyroid phenotypes.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>By combining TALENs injections with a targeting construct that contained a <jats:italic>gfp<\/jats:italic> transgene, we were able to recover an allele in which GFP is expressed in the <jats:italic>nkx2.4b<\/jats:italic> expression domains, with homozygous phenotypes suggesting the creation of a loss\u2010of\u2010function transgenic line. These results demonstrate the creation of a useful tool for studying hypothalamus and thyroid development.<\/jats:p><\/jats:sec>","DOI":"10.1002\/dvdy.224","type":"journal-article","created":{"date-parts":[[2020,7,9]],"date-time":"2020-07-09T09:06:36Z","timestamp":1594285596000},"page":"1387-1393","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A transgene targeted to the zebrafish <i>nkx2.4b<\/i> locus drives specific <scp>green fluorescent protein<\/scp> expression and disrupts thyroid development"],"prefix":"10.1002","volume":"249","author":[{"given":"David A.","family":"Hutcheson","sequence":"first","affiliation":[{"name":"Department of Neurobiology and Anatomy University of Utah  Salt Lake City Utah USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9796-7514","authenticated-orcid":false,"given":"Yuanyuan","family":"Xie","sequence":"additional","affiliation":[{"name":"Department of Molecular, Cellular and Developmental Biology, BioFrontiers Institute University of Colorado  Boulder Colorado USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Priscilla","family":"Figueroa","sequence":"additional","affiliation":[{"name":"Department of Neurobiology and Anatomy University of Utah  Salt Lake City Utah USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8037-6694","authenticated-orcid":false,"given":"Richard I.","family":"Dorsky","sequence":"additional","affiliation":[{"name":"Department of Neurobiology and Anatomy University of Utah  Salt Lake City Utah 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