PUBLICATION
A transgene targeted to the zebrafish nkx2.4b locus drives specific GFP expression and disrupts thyroid development
- Authors
- Hutcheson, D.A., Xie, Y., Figueroa, P., Dorsky, R.I.
- ID
- ZDB-PUB-200710-23
- Date
- 2020
- Source
- Developmental Dynamics : an official publication of the American Association of Anatomists 249(11): 1387-1393 (Journal)
- Registered Authors
- Dorsky, Richard, Hutcheson, David, Xie, Yuanyuan
- Keywords
- Development, GFP, Hypothalamus, Nkx2.4b, TALENs, Thyroid, transgene
- MeSH Terms
-
- Animals
- Animals, Genetically Modified*/embryology
- Animals, Genetically Modified*/genetics
- Gene Expression Regulation, Developmental*
- Green Fluorescent Proteins*/biosynthesis
- Green Fluorescent Proteins*/genetics
- Homeodomain Proteins/genetics*
- Thyroid Gland/embryology*
- Transgenes*
- Zebrafish*/embryology
- Zebrafish*/genetics
- Zebrafish Proteins/genetics*
- PubMed
- 32644242 Full text @ Dev. Dyn.
Citation
Hutcheson, D.A., Xie, Y., Figueroa, P., Dorsky, R.I. (2020) A transgene targeted to the zebrafish nkx2.4b locus drives specific GFP expression and disrupts thyroid development. Developmental Dynamics : an official publication of the American Association of Anatomists. 249(11):1387-1393.
Abstract
Background With the goal of labeling and manipulating the zebrafish hypothalamus, we sought to target a gfp transgene to the expression domains of nkx2.4b, a gene expressed during hypothalamic and thyroid development. We combined 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. This article is protected by copyright. All rights reserved.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping