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Blood, 15 June 2005, Vol. 105, No. 12, pp. 4657-4663.
Prepublished online as a Blood First Edition Paper on March 3, 2005; DOI 10.1182/blood-2004-09-3554.
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Submitted September 14, 2004
Accepted February 19, 2005
A high level of endothelial cell-specific gene expression by a combination of 5' flanking region and 5' half of the first intron of VE-cadherin gene
Hiroshi Hisatsune, Kazuyoshi Matsumura, Minetaro Ogawa, Akiyoshi Uemura, Nobuyuki Kondo, Jun K Yamashita, Hideto Katsuta, Satomi Nishikawa, Tsutomu Chiba, and Shin-Ichi Nishikawa*
Division of Gastroenterology and Hepatology, Department of Internal Medicine, Graduate School of Medicine, Kyoto University, Kyoto, Kyoto, Japan
Department of Cell Differentiation, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Kumamoto, Japan
Stem Cell Biology Laboratory, Center for Developmental Biology, Kobe, Hyogo, Japan
Department of Thoracic Surgery, Kyoto University, Kyoto, Kyoto, Japan
Laboratory of Stem Cell Differentiation, Stem Cell Research Center, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Kyoto, Japan
Department of Ophthalmology and Visual Sciences, Kyoto University, Kyoto, Kyoto, Japan
* Corresponding author; email: nishikawa{at}cdb.riken.jp.
In order to develop a tool to obtain a high level of gene expression specifically in endothelial cells (EC), we assessed enhancer activity of fragments in the first intron of VE-cadherin gene using three different experimental systems; luciferase assay in F2 EC line, GFP expression in ECs generated in ES cell differentiation culture, and GFP expression in transgenic mice. While 2.5kbp 5' flanking sequence of VE-cadherin gene is EC specific, addition of 4kbp 5' half of the first intron effected an enhancement of the gene expression level in all three assay systems. No other fragments tested in this study could confer such effects. Comparing with other gene expression unit, the unit described in this study would be the most optimum one that is available to date for EC specific gene expression. As this unit can express gene in VE-cadherin+ progenitors of hematopoietic cells but not in fully committed hematopoietic cells, it will be useful to manipulate specifically the uncommitted progenitor stage during hematopoietic cell differentiation.
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