Generation of an anti-fibrotic therapeutic gene using a tubular epithelium-specific expression gene casette and iKB cDNA
使用肾小管上皮特异性表达基因盒和 iKB cDNA 生成抗纤维化治疗基因
基本信息
- 批准号:14571033
- 负责人:
- 金额:$ 1.73万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:2002
- 资助国家:日本
- 起止时间:2002 至 2004
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
To identify tubular epithelium-specific, positive regulatory elements in the promoter of osteopontin (OPN) gene, we generated luciferase reporter gene constructs bearing various fragments of the OPN promoter and performed transient transfection experiments using murine tubular epithelial cells (mPTEC). In addition to mPTEC, fibroblasts (3T3) and monocytes (RAW264) were also employed for the transfection experiments. The -600 to -825 bp fragment of the OPN promoter constitutively showed positive transcription activity in mPTEC, but not in 3T3 and RAW264. In contrast, the -115 to -189 bp fragment showed positive transcription activity in mPTEC as well as 3T3 and RAW264 in response to recombinant transforming growth factor-b1 treatment. Thus, the -600 to -825 bp fragment is suggested to contain tubular epithelium-specific, positive regulatory elements. We are now generating tubular epithelium-specific expression gene casette using this fragment, and testing whether tubular epithelium-specific expression of iKB can attenuate fibrosis through down-regulation of NF-kB activity in vitro and in vivo.
为了鉴定骨桥蛋白(OPN)基因启动子中的肾小管上皮特异性阳性调控元件,我们构建了带有OPN启动子各种片段的荧光素酶报告基因构建体,并使用小鼠肾小管上皮细胞(mPTEC)进行瞬时转染实验。除了mPTEC之外,成纤维细胞(3 T3)和单核细胞(RAW 264)也用于转染实验。OPN启动子的-600至-825 bp片段在mPTEC中组成性地显示出阳性转录活性,但在3 T3和RAW 264中不显示。相反,-115至-189 bp片段在mPTEC以及3 T3和RAW 264中显示出对重组转化生长因子-b1处理的响应的阳性转录活性。因此,-600至-825 bp片段被认为含有肾小管上皮特异性的正调控元件。我们现在正在使用该片段产生肾小管上皮特异性表达基因盒,并测试肾小管上皮特异性表达iKB是否可以通过下调体外和体内NF-κ B活性来减轻纤维化。
项目成果
期刊论文数量(2)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Direct contact between human peripheral blood mononuclear cells and renal fibroblasts facilitates the expression of monocyte chemoattractant protein-1
- DOI:10.1159/000071480
- 发表时间:2003-07-01
- 期刊:
- 影响因子:4.2
- 作者:Hao, LR;Okada, H;Suzuki, H
- 通讯作者:Suzuki, H
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