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
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
复制标题
DOI：
10.1182/blood-2004-09-3554
复制
发表时间：
2005-06-15
期刊：
影响因子：
20.3
通讯作者：
Nishikawa, S
中科院分区：
文献类型：
--
作者：
Hisatsune, H;Matsumura, K;Nishikawa, S
为了开发一种工具，以获得高水平的基因表达，特别是在内皮细胞（EC），我们评估了增强子活性的片段在第一内含子的VE-钙粘蛋白基因使用3种不同的实验系统：荧光素酶测定在F2 EC线，绿色荧光蛋白（GFP）表达在胚胎干细胞（ES）分化培养中产生的EC，和GFP表达在转基因小鼠。尽管VE-钙粘蛋白基因的2.5-kbp（千碱基对）5'侧翼序列是EC特异性的，但在第一内含子的5'半中添加4kbp影响了所有3种测定系统中基因表达水平的增强。本研究中测试的其他碎片都无法产生此类效果。与其他基因表达单位相比，本研究中描述的单位将是迄今为止EC特异性基因表达的最佳单位。由于该单位可以在造血细胞的VE-钙粘蛋白（+）祖细胞中表达基因，但在完全定型的造血细胞中不表达，因此在造血细胞分化期间特异性地操纵未定型祖细胞阶段将是有用的。(c)2005年，美国血液学会。
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 (kilo-base 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 hematopoletic cell differentiation. (c) 2005 by The American Society of Hematology.