Regulation of CD154 gene expression in vivo

描述（由申请人提供）：活化的CD 154（CD 40配体）表达 CD 154-CD 40相互作用在模型中显示出有益的作用， 自身免疫炎症和移植排斥反应尽管如此， 他知道它的表达调控，除了它似乎有 有几个特征将其与细胞因子基因区分开来。几项研究 这表明mRNA稳定性作为调控的主要决定因素的重要性。 CD 154表达。我们实验室的工作描绘了一个新的区域， 调节嵌合报告基因表达的人CD 154 >UTR。这 顺式作用元件在鼠和人细胞中都有活性， 保守的，并介导相当于富含Au元素（AURE）的效应 其已被证明是调节细胞因子的中心， TNF-α和IL-2。仔细分析正常T淋巴细胞中mRNA的稳定性， 受技术因素限制。因此，我们建议测试 这种顺式作用元件在CD 154基因表达中的作用是通过同源突变来实现的。 在小鼠生殖系中进行重组以产生小鼠系，其中该重组在小鼠生殖系中进行。 元素不再起作用。通过这种方法，我们将确定 CD 154表达的转录后调节在健康和 疾病，正如用TNF-α AURE突变小鼠令人信服地证明的那样。 因此，我们将创建一个模型系统，以研究其独特的 监管，并可能开发新的方法来监管其 生物合成

DESCRIPTION (provided by applicant): CD154 (CD40 ligan) expression by activated of CD 154-CD40 interactions has been shown to have salutary effects in models of autoimmunity, inflammation, and graft rejection. Despite this, little is known about the regulation of its expression, except that it appears to have several features that distinguish it from cytokine genes. Several studies have indicated the importance of mRNA stability as a major determinant in regulating CD154 expression. Work in our laboratory has delineated a novel region of the human CD 154 >UTR that regulates chimeric reporter gene expression. This cis-acting element is active in both murine and human cells, is highly conserved, and mediates effects equivalent to that of AU-rich elements (AURE) which have been shown to be central in the regulation of cytokines such as TNF-a and IL-2. Careful analysis of mRNA stability in normal T lymphocytes is limited by technical considerations. Therefore, we propose to test the role of this cis-acting element in CD 154 gene expression by mutating it by homologous recombination in the murine germ line to generate a mouse line in which this element is no longer functional. Through this approach, we will determine the role of post-transcriptional regulation of CD 154 expression in health and disease, as was compellingly demonstrated with the TNF-alpha AURE mutant mice. As a result, we will create a model system with which to study its unique regulation and possibly develop novel approaches to regulating its biosynthesis.