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Title

CONSTRUCTING A MOUSE OCT4 PROMOTER/EGFP VECTOR, AS A WHOLE-CELLULAR REPORTER TO MONITOR THE PLURIPOTENT STATE OF CELLS

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 Start Page 2 | End Page 9

Abstract

 Background: 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.Methods: 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.Results: 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.Conclusion: 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.

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