A transgene targeted to the zebrafish nkx2.4b locus drives specific green fluorescent protein expression and disrupts thyroid development.
Dev Dyn
; 249(11): 1387-1393, 2020 11.
Article
em En
| MEDLINE
| ID: mdl-32644242
ABSTRACT
BACKGROUND:
With the goal of labeling and manipulating the zebrafish hypothalamus, we sought to target a green fluorescent protein (gfp) transgene to the expression domains of nkx2.4b, a gene expressed during hypothalamic and thyroid development. We combined transcription activator-like effector nucleases (TALENs)-mediated mutagenesis with a targeting construct to enable insertion of a gfp transgene into the endogenous nkx2.4b genomic locus.RESULTS:
Injection of TALENs targeted to the first exon of nkx2.4b 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 gfp 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 nkx2.4b and exhibited hypothyroid phenotypes.CONCLUSIONS:
By combining TALENs injections with a targeting construct that contained a gfp transgene, we were able to recover an allele in which GFP is expressed in the nkx2.4b expression domains, with homozygous phenotypes suggesting the creation of a loss-of-function transgenic line. These results demonstrate the creation of a useful tool for studying hypothalamus and thyroid development.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Glândula Tireoide
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Peixe-Zebra
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Animais Geneticamente Modificados
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Proteínas de Homeodomínio
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Regulação da Expressão Gênica no Desenvolvimento
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Transgenes
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Proteínas de Peixe-Zebra
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Proteínas de Fluorescência Verde
Limite:
Animals
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article