A transgene targeted to the zebrafish nkx2.4b locus drives specific green fluorescent protein expression and disrupts thyroid development.
A transgene targeted to the zebrafish nkx2.4b locus drives specific green fluorescent protein expression and disrupts thyroid development.
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DOI：
10.1002/dvdy.224
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发表时间：
2020-11
期刊：
影响因子：
--
通讯作者：
Dorsky RI
中科院分区：
文献类型：
--
作者：
Hutcheson DA;Xie Y;Figueroa P;Dorsky RI
为了标记和操纵斑马鱼下丘脑，我们试图将gfp转基因靶向nkx2.4b的表达域，nkx2.4b是下丘脑和甲状腺发育过程中表达的基因。我们将TALENs介导的突变与靶向构建相结合，使gfp转基因能够插入内源性nkx2.4b基因组位点。注射靶向nkx24 b的第一个外显子的TALENs产生了一个预测的零等位基因，纯合突变胚胎显示甲状腺标志物的缺失。从同时注射了TALENs和携带gfp转基因的靶向构建体的胚胎中，我们恢复了gfp在下丘脑和甲状腺中特异性表达的细胞系。该等位基因纯合的鱼缺少nkx2.4b的外显子1，表现出甲状腺功能低下的表型。通过将TALENs注射剂与含有gfp转基因的靶向构建物结合，我们能够恢复gfp在nkx24 b表达域表达的等位基因，纯合表型表明创建了一个功能缺失的转基因系。这些结果为研究下丘脑和甲状腺发育提供了一个有用的工具。
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. 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. 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.
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影响因子：
11.8
作者：
Wang, Xu;Kopinke, Daniel;Lin, Junji;McPherson, Adam D.;Duncan, Robert N.;Otsuna, Hideo;Moro, Enrico;Hoshijima, Kazuyuki;Grunwald, David J.;Argenton, Francesco;Chien, Chi-Bin;Murtaugh, L. Charles;Dorsky, Richard I.
通讯作者：
Dorsky, Richard I.
影响因子：
5.3
作者：
MIZUNO, K;GONZALEZ, FJ;KIMURA, S
通讯作者：
KIMURA, S
影响因子：
--
作者：
Opitz, Robert;Maquet, Emilie;Costagliola, Sabine
通讯作者：
Costagliola, Sabine
影响因子：
16.2
作者：
Rohr, KB;Barth, KA;Wilson, SW
通讯作者：
Wilson, SW
DOI：
10.1016/j.beem.2013.08.005
发表时间：
2014-03-01
影响因子：
7.4
作者：
Szinnai, Gabor
通讯作者：
Szinnai, Gabor