Generation of Mouse for Conditional Expression of Forkhead Box A2.
Generation of Mouse for Conditional Expression of Forkhead Box A2.
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DOI：
10.1210/en.2018-00158
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发表时间：
2018-04
期刊：
影响因子：
4.8
通讯作者：
Peng Wang;San-pin Wu;K. Brooks;A. M. Kelleher;Jessica Milano-Foster;F. DeMayo;T. Spencer
中科院分区：
文献类型：
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作者：
Peng Wang;San-pin Wu;K. Brooks;A. M. Kelleher;Jessica Milano-Foster;F. DeMayo;T. Spencer
叉头盒A2（FOXA 2）是参与器官发育、功能和癌症的先驱转录因子。在子宫中，FOXA 2是怀孕所必需的，并在子宫内膜的腺体中特异性表达。FOXA 2功能的丧失发生在人类子宫内膜癌的发展过程中。目前的研究描述了小鼠FOXA 2的条件表达的小鼠模型的发展。使用由位于Rosa 26位点的小基因组成的系统，我们在胚胎干细胞中产生了CAG-S-mFOXA 2等位基因，随后在小鼠中产生了CAG-S-mFOXA 2等位基因;在激活之前，由于启动子和转基因之间插入了floxed终止盒，所以小基因是沉默的。为了验证功能性，将具有CAG-S-mFOXA 2等位基因的小鼠与分别在未成熟和成年子宫中表达Cre的孕酮受体（Pgr）-Cre小鼠和乳转铁蛋白（Ltf）-iCre小鼠杂交。在未成熟的Pgr-Cre-CAG-S-mFoxa 2小鼠中，FOXA 2蛋白在子宫腔上皮（LE）、腺上皮（GE）、基质和子宫肌层的内层中表达。有趣的是，在成年Pgr-Cre-CAG-S-mFoxa 2小鼠子宫的大部分LE中未观察到FOXA 2蛋白，尽管FOXA 2维持在基质、GE和子宫肌层中。成年Pgr-Cre-CAG-S-mFoxa 2雌性完全不育。相比之下，Ltf-iCre-CAG-S-mFoxa 2小鼠具有生育能力，子宫中没有可检测的组织学差异。Pgr-Cre-CAG-S-mFoxa 2小鼠的成年子宫较小，包含很少的子宫内膜腺体，并显示部分分层的LE和GE区域。该转基因小鼠系为阐明和探索FOXA 2的功能提供了宝贵的资源。
Forkhead box A2 (FOXA2) is a pioneer transcription factor involved in organ development, function, and cancer. In the uterus, FOXA2 is essential for pregnancy and expressed specifically in the glands of the endometrium. Loss of FOXA2 function occurs during development of endometrial cancer in humans. The current study describes the development of a mouse model for conditional expression of mouse FOXA2. Using a system consisting of a minigene located at the Rosa26 locus, we generated a CAG-S-mFOXA2 allele in embryonic stem cells and subsequently in mice; before activation, the minigene is silent because of a floxed stop cassette inserted between the promoter and the transgene. To validate functionality, mice with the CAG-S-mFOXA2 allele were crossed with progesterone receptor (Pgr)-Cre mice and lactotransferrin (Ltf)-iCre mice that express Cre in the immature and adult uterus, respectively. In immature Pgr-Cre-CAG-S-mFoxa2 mice, FOXA2 protein was expressed in the luminal epithelium (LE), glandular epithelium (GE), stroma, and inner layer of the myometrium. Interestingly, FOXA2 protein was not observed in most of the LE of uteri from adult Pgr-Cre-CAG-S-mFoxa2 mice, although FOXA2 was maintained in the stroma, GE, and myometrium. The adult Pgr-Cre-CAG-S-mFoxa2 females were completely infertile. In contrast, Ltf-iCre-CAG-S-mFoxa2 mice were fertile with no detectable histological differences in the uterus. The adult uterus of Pgr-Cre-CAG-S-mFoxa2 mice was smaller, contained few endometrial glands, and displayed areas of partially stratified LE and GE. This transgenic mouse line is a valuable resource to elucidating and exploring FOXA2 function.