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.
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.
Hisatsune
Hiroshi
H
Division of Gastroenterology and Hepatology, Department of Internal Medicine, Graduate School of Medicine, Stem Cell Research Center, Kyoto University, Japan.
Matsumura
Kazuyoshi
K
Ogawa
Minetaro
M
Uemura
Akiyoshi
A
Kondo
Nobuyuki
N
Yamashita
Jun K
JK
Katsuta
Hideto
H
Nishikawa
Satomi
S
Chiba
Tsutomu
T
Nishikawa
Shin-ichi
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D015703:Antigens, CD; D015820:Cadherins / Q000096:biosynthesis* / Q000235:genetics*; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D004742:Enhancer Elements, Genetic; D005434:Flow Cytometry; D017930:Genes, Reporter; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology; D007438:Introns; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D010957:Plasmids / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology; D000099152:Cadherin 5
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D015820:Cadherins; D049452:Green Fluorescent Proteins; D008156:Luciferases; D000099152:Cadherin 5