Constructing a Mouse Oct4 Promoter/EGFP Vector, as a Whole-Cellular Reporter to Monitor the Pluripotent State of Cells

• Published in: Avicenna journal of medical biotechnology Vol. 5; no. 1; pp. 2 - 9
• Main Authors: 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
• Format: Journal Article
• Language: English
• Published: Iran Avicenna Research Institute 01.01.2013
• Subjects: Fluorescence; Infection; Original; Stem cells; Transcription factors; Mice; Embryonic stem cells; Enhanced green fluorescent protein
• ISSN: 2008-2835; 2008-4625

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.