Payer Bernhard, Chuva de Sousa Lopes Susana M, Barton Sheila C, Lee Caroline, Saitou Mitinori, Surani M Azim
Wellcome Trust/Cancer Research UK Gurdon Institute of Cancer and Developmental Biology, University of Cambridge, Cambridge, United Kingdom.
Genesis. 2006 Feb;44(2):75-83. doi: 10.1002/gene.20187.
The relationship between germ cells and pluripotent embryonic stem (ES) cells is of particular interest, together with approaches to generate primordial germ cell (PGCs) from ES cells. A critical requirement in these experiments is the ability to unambiguously detect PGCs with the use of, for example, reporter genes. The currently available transgenic reporters do not show exclusive expression in PGCs at their earliest developmental stages. Here we describe the use of germline-restricted expression of stella, which is currently the best marker gene for PGCs. We generated two stella-GFP reporters and show that both transgenes surpass other reporters in terms of timing and specificity of expression in PGCs. Additionally, we demonstrate the usefulness of stella-GFP during the derivation of PGCs from ES cells.
生殖细胞与多能胚胎干细胞(ES细胞)之间的关系备受关注,同时也包括从ES细胞生成原始生殖细胞(PGC)的方法。这些实验的一个关键要求是能够使用例如报告基因明确检测PGC。目前可用的转基因报告基因在PGC发育的最早阶段并未表现出在PGC中的特异性表达。在此,我们描述了使用生殖系特异性表达的斯泰拉(Stella),它是目前PGC的最佳标记基因。我们构建了两个斯泰拉 - 绿色荧光蛋白(Stella-GFP)报告基因,并表明这两个转基因在PGC表达的时间和特异性方面都优于其他报告基因。此外,我们证明了Stella-GFP在从ES细胞衍生PGC过程中的实用性。