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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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HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY

High level of endothelial cell-specific gene expression by a combination of the 5' flanking region and the 5' half of the first intron of the 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

From the Division of Gastroenterology and Hepatology, the Department of Internal Medicine, Graduate School of Medicine, the Department of Thoracic Surgery, the Laboratory of Stem Cell Differentiation, Stem Cell Research Center, Institute for Frontier Medical Sciences, and the Department of Ophthalmology and Visual Sciences, Kyoto University; the Department of Cell Differentiation, Institute of Molecular Embryology and Genetics, Kumamoto University; and the Stem Cell Biology Laboratory, Center for Developmental Biology, Kobe, Japan.

To develop a tool to obtain a high level of gene expression specifically in endothelial cells (ECs), we assessed enhancer activity of fragments in the first intron of the VE-cadherin gene using 3 different experimental systems: luciferase assay in the F2 EC line, green fluorescent protein (GFP) expression in ECs generated in embryonic stem (ES) cell differentiation culture, and GFP expression in transgenic mice. Although the 2.5-kbp (kilobase pair) 5' flanking sequence of the VE-cadherin gene is EC specific, adding 4 kbp of the 5' half of the first intron affected an enhancement of the gene expression level in all 3 assay systems. No other fragments tested in this study could confer such effects. Compared with other gene expression units, the unit described in this study would be the most optimum one available to date for EC-specific gene expression. Because this unit can express genes 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. (Blood. 2005; 105:4657-4663)


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