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This Article Free via Open Access: OA OA Full Text Full Text (PDF) Supplemental Data OA All Versions of this Article: 2007-0902v1 26/7/1841    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints/Permissions Google Scholar Articles by Zhang, X. Articles by Gottlieb, D. I. PubMed PubMed Citation Articles by Zhang, X. Articles by Gottlieb, D. I.

EMBRYONIC STEM CELLS

Embryonic Stem Cells as a Platform for Analyzing Neural Gene Transcription

Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, Missouri, USA

Key Words. Gene targeting • Olig2 • Transcription • Embryonic stem cells • Neural differentiation • Promoter

Correspondence: David I. Gottlieb, Ph.D., Department of Anatomy and Neurobiology, Box 8108, Washington University School of Medicine, 660 S. Euclid Avenue, St. Louis, Missouri 63110, USA. Telephone: 314-362-2758; Fax: 314-362-3446; e-mail: gottlied{at}pcg.wustl.edu

Received October 26, 2007; accepted for publication April 12, 2008.
First published online in STEM CELLS EXPRESS   April 24, 2008.

There is a need for improved methods to analyze transcriptional control of mammalian stem cell genes. We propose that embryonic stem cells (ESCs) will have broad utility as a model system, because they can be manipulated genetically and then differentiated into many cell types in vitro, avoiding the need to make mice. Results are presented demonstrating the utility of ESCs for analyzing cis-acting sequences using Olig2 as a model gene. Olig2 is a transcription factor that plays a key role in the development of a ventral compartment of the nervous system and the oligodendrocyte lineage. The functional role of an upstream region (USR) of the Olig2 gene was investigated in ESCs engineered at the undifferentiated stage and then differentiated into ventral neural cells with sonic hedgehog and retinoic acid. Deletion of the USR from the native gene via gene targeting eliminates expression in ventral neural cells differentiated in cell culture. The USR is also essential for regulated expression of an Olig2 transgene inserted at a defined foreign chromosomal site. A subregion of the USR has nonspecific promoter activity in transient transfection assays in cells that do not express Olig2. Taken together, the data demonstrate that the USR contains a promoter for the Olig2 gene and suggest that repression contributes to specific expression. The technology used in this study can be applied to a wide range of genes and cell types and will facilitate research on cis-acting DNA elements of mammalian genes.

Disclosure of potential conflicts of interest is found at the end of this article.