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
  • Zebrafish*/embryology
  • Zebrafish*/genetics
  • Thyroid Gland/embryology*
  • Animals
  • Homeodomain Proteins/genetics*
  • Animals, Genetically Modified*/embryology
  • Animals, Genetically Modified*/genetics
  • Gene Expression Regulation, Developmental*
  • Transgenes*
  • Green Fluorescent Proteins*/biosynthesis
  • Green Fluorescent Proteins*/genetics
  • Zebrafish Proteins/genetics*
(all 12)
PubMed
32644242 Full text @ Dev. Dyn.
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
Figures
Figure Gallery (3 images)
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Expression
Phenotype
Fish Conditions Stage Phenotype Figure
nkx2.4bzd20/zd20standard conditionsLong-pec
nkx2.4bzd20/zd20standard conditionsLong-pec
nkx2.4bzd20/zd20standard conditionsLong-pec
nkx2.4bzd20/zd20standard conditionsLong-pec
nkx2.4bzd21Tg/zd21Tgstandard conditionsDays 30-44
nkx2.4bzd21Tg/zd21Tgstandard conditionsDays 30-44
1 - 6 of 6
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Mutations / Transgenics
Allele Construct Type Affected Genomic Region
zd20
    		Small Deletion
    zd21TgTransgenic Insertion
    1 - 2 of 2
    Show
    Human Disease / Model
    No data available
    Sequence Targeting Reagents
    Target Reagent Reagent Type
    nkx2.4bTALEN2-nkx2.4bTALEN
    1 - 1 of 1
    Show
    Fish
    Fish
    nkx2.4bzd20/zd20
    nkx2.4bzd21Tg/zd21Tg
    WT
    1 - 3 of 3
    Show
    Antibodies
    No data available
    Orthology
    No data available
    Engineered Foreign Genes
    Marker Marker Type Name
    EGFPEFGEGFP
    GFPEFGGFP
    1 - 2 of 2
    Show
    Mapping
    No data available
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    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.