ARTICLE

Cell Research, 15(5):317-324, May 2005

Isolation and characterization of the murine Nanog gene promoter

Da Yong WU^*,  Zhen YAO

Laboratory of Molecular Cell Biology, Laboratory of Stem Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China.

Received, Mar 5, 2005 Revised, Mar 12 , 2005 Accepted, Mar 21 , 2005

Correspondence: Da Yong WU +86-21-54921366 (phone) +86-21-54921366 (fax) dywu@sibs.ac.cn

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Nanog protein is expressed in the interior cells of compacted morulae and maintained till epiblasts but downregulated by implantation stage. It is also expressed in embryonic stem cells, embryonic carcinoma cells and embryonic germ cells but disappeared in differentiated ES cells. In this study, we have isolated, sequenced, and performed the first characterization of the Nanog promoter. The transcription start sites were mapped by primer extension analysis. Two promoter regions were found upstream the transcription start sites and the expression of major Nanog promoter/reporter gene construct is abolished in differentiated F9 EC cells as compared to the undifferentiated counterpart. We also showed that a putative octamer motif (ATGCAAAA) is necessary for the major promoter activity. Gel shift and supershift assays showed that Oct-1, Oct-4 and Oct-6 protein selectively bind to the octamer motif.

Keywords: Nanog, promoter, F9 EC cells, Oct-1, Oct-4, Oct-6.