Ghorbani Reza, Emamzadeh Abdolrahman, Khazaie Yahya, Dormiani Kianoush, Ghaedi Kamran, Rabbani Mohammad, Foruzanfar Mahboubeh, Karbalaie Khadijeh, Karamali Fereshteh, Lachinani Liana, Kiani-Esfahani Abbas, Nematollahi Marzieh, Esfahani Mohammad Hossein Nasr
Department of Molecular Biotechnology, Cell Sciences Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran ; Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan, Iran.
Avicenna J Med Biotechnol. 2013 Jan;5(1):2-9.
The transcription factor Oct-4, is an important marker of undifferentiating level and a key regulating factor for maintenance of pluripotency in cells. Establishment of an Oct-4 promoter-based reporter system is an appropriate tool for monitoring the differentiation of embryonic stem cells both in vivo and in vitro.
In the present study, we report construction of a recombinant vector, pDB2 Oct4 promoter/EGFP, in which expression of Enhanced Green Fluorescent Protein (EGFP) was controlled by the mouse Oct-4 promoter.
In transfected mouse embryonic stem cells with this vector, EGFP was predicted to be specifically expressed in pluripotency state. After transfection, high-level expression of EGFP under the control of Oct-4 promoter was observed in manipulated embryonic stem cells.
Thus, our new cellular reporter showed that both the properties of embryonic cells and expression the EGFP could be of great help in studying the differentiating and reprogramming mechanisms of mESCs.
转录因子Oct-4是未分化水平的重要标志物，也是维持细胞多能性的关键调控因子。建立基于Oct-4启动子的报告系统是在体内和体外监测胚胎干细胞分化的合适工具。
在本研究中，我们报告了重组载体pDB2 Oct4启动子/EGFP的构建，其中增强型绿色荧光蛋白（EGFP）的表达由小鼠Oct-4启动子控制。
用该载体转染小鼠胚胎干细胞后，预测EGFP会在多能状态下特异性表达。转染后，在操作的胚胎干细胞中观察到Oct-4启动子控制下EGFP的高水平表达。
因此，我们的新细胞报告系统表明，胚胎细胞的特性和EGFP的表达对研究小鼠胚胎干细胞的分化和重编程机制有很大帮助。
