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用于滋养层特异性基因操作的Elf5-Cre基因敲入小鼠品系的构建

Generation of Elf5-Cre knockin mouse strain for trophoblast-specific gene manipulation.

作者信息

Kong Shuangbo, Liang Guixian, Tu Zhaowei, Chen Dunjin, Wang Haibin, Lu Jinhua

机构信息

Reproductive Medical Center, The First Affiliated Hospital of Xiamen University, Xiamen, Fujian, 361003, People's Republic of China.

Fujian Provincial Key Laboratory of Reproductive Health Research, Medical College of Xiamen University, Xiamen, Fujian, 361102, People's Republic of China.

出版信息

Genesis. 2018 Apr;56(4):e23101. doi: 10.1002/dvg.23101. Epub 2018 Mar 25.

Abstract

Placental development is a complex and highly controlled process during which trophoblast stem cells differentiate to various trophoblast subtypes. The early embryonic death of systemic gene knockout models hampers the investigation of these genes that might play important roles during placentation. A trophoblast specific Cre mouse model would be of great help for dissecting out the potential roles of these genes during placental development. For this purpose, we generate a transgenic mouse with the Cre recombinase inserted into the endogenous locus of Elf5 gene that is expressed specifically in placental trophoblast cells. To analyze the specificity and efficiency of Cre recombinase activity in Elf5-Cre mice, we mated Elf5-Cre mice with Rosa26 reporter mice, and found that Elf5-Cre transgene is expressed specifically in the trophoectoderm as early as embryonic day 4.5 (E4.5). By E12.5, the activity of Elf5-Cre transgene was detected exclusively in all derivatives of trophoblast lineages, including spongiotrophoblast, giant cells, and labyrinth trophoblasts. In addition, Elf5-Cre transgene was also active during spermatogenesis, from spermatids to mature sperms, which is consistent with the endogenous Elf5 expression in testis. Collectively, our results provide a unique tool to delete specific genes selectively and efficiently in trophoblast lineage during placentation.

摘要

胎盘发育是一个复杂且高度受控的过程,在此过程中,滋养层干细胞分化为各种滋养层亚型。系统性基因敲除模型的早期胚胎死亡阻碍了对这些可能在胎盘形成过程中发挥重要作用的基因的研究。滋养层特异性Cre小鼠模型对于剖析这些基因在胎盘发育过程中的潜在作用将有很大帮助。为此,我们构建了一种转基因小鼠,将Cre重组酶插入到Elf5基因的内源性位点,该基因在胎盘滋养层细胞中特异性表达。为了分析Elf5-Cre小鼠中Cre重组酶活性的特异性和效率,我们将Elf5-Cre小鼠与Rosa26报告基因小鼠交配,发现Elf5-Cre转基因早在胚胎第4.5天(E4.5)就在滋养外胚层中特异性表达。到E12.5时,Elf5-Cre转基因的活性仅在滋养层谱系的所有衍生物中检测到,包括海绵滋养层、巨细胞和迷路滋养层。此外,Elf5-Cre转基因在精子发生过程中也有活性,从精子细胞到成熟精子,这与睾丸中内源性Elf5的表达一致。总的来说,我们的结果提供了一种独特的工具,可在胎盘形成过程中选择性地和有效地在滋养层谱系中删除特定基因。

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