Generation of a germ cell-specific mouse transgenic CHERRY reporter, Sohlh1-mCherryFlag.
Generation of a germ cell-specific mouse transgenic CHERRY reporter, Sohlh1-mCherryFlag.
复制标题
DOI：
10.1002/dvg.22347
复制
发表时间：
2013-01
期刊：
影响因子：
1.5
通讯作者：
Rajkovic, Aleksandar
中科院分区：
文献类型：
--
作者：
Suzuki, Hitomi;Dann, Christina Tenenhaus;Rajkovic, Aleksandar
对分化生殖细胞的可视化对于了解卵巢中原始卵泡的形成以及精原干细胞对分化的承诺至关重要。我们在Sohlh1启动子的指导下，成功地构建了BAC转基因小鼠品系Sohlh1-mCherryFlag(S1CF)。Sohlh1是一个生殖细胞特异性基因，编码基本螺旋-环-螺旋(BHLH)转录调节因子，在卵子发生和精子发生中是必不可少的。Sohlh1的表达是独特的，仅限于围产期和早期卵泡卵母细胞和分化中的精原细胞。Sohlh1-mCherryFlag转基因是为了将SOHLH1与带有3-串联FLAG标签的红色荧光蛋白Cherry融合在一起。S1CF动物特异地在围产期卵巢的卵母细胞和成年卵巢的小卵泡以及精原细胞中发出荧光，这种模式类似于内源性的SOHLH1。此外，S1CF还挽救了雄性和雌性Sohlh1−/−动物的生殖细胞丢失和不育。S1CF上的FLAG标记可用于免疫染色和免疫沉淀。Sohlh1-mCherryFlag转基因小鼠为研究早期生殖细胞分化以及体内成像和分化生殖细胞的纯化提供了独特的模型。
Visualization of differentiating germ cells is critical to understanding the formation of primordial follicles in the ovary, and the commitment of spermatogonial stem cells to differentiation. We engineered and generated a BAC transgenic mouse line, Sohlh1-mCherryFlag (S1CF), under the direction of the native Sohlh1 promoter. Sohlh1 is a germ cell-specific gene that encodes the basic helix-loop-helix (bHLH) transcriptional regulator that is essential in oogenesis and spermatogenesis. Sohlh1 expression is unique, and is limited to perinatal and early follicle oocytes and differentiating spermatogonia. The Sohlh1-mCherryFlag transgene was engineered to fuse SOHLH1 to the red fluorescent protein CHERRY with 3-tandem-FLAG tags. S1CF animals fluoresce specifically in the oocytes of perinatal ovaries and small follicles in adult ovaries, as well as in spermatogonia, a pattern that is similar to endogenous SOHLH1. Moreover, S1CF rescued germ cell loss and infertility in both male and female Sohlh1−/− animals. The FLAG-tag on S1CF was effective for immunostaining and immunoprecipitation. The Sohlh1-mCherryFlag transgenic mouse provides a unique model to study early germ cell differentiation, as well as in vivo imaging and purification of differentiating germ cells.
登录
查看更多内容
影响因子：
14.9
作者：
Warming S;Costantino N;Court DL;Jenkins NA;Copeland NG
通讯作者：
Copeland NG
影响因子：
5.3
作者：
Hoodbhoy, Tanya;Aviles, Manuel;Dean, Jurrien
通讯作者：
Dean, Jurrien
影响因子：
4.6
作者：
Molyneaux, KA;Zinszner, H;Lehmann, R
通讯作者：
Lehmann, R
影响因子：
4.6
作者：
Takeuchi, Y;Molyneaux, K;Wylie, C
通讯作者：
Wylie, C
影响因子：
2.7
作者：
Ballow, D.;Meistrich, M. L.;Rajkovic, A.
通讯作者：
Rajkovic, A.