Knock-in of Posttranslational Mutations of Nuclear Receptor Coregulator Genes

我们的长期目标是促进识别协调制子的PTM的个体和组合角色 在分化、发育、繁殖和代谢动态平衡方面使用小鼠作为 最终的模式。这一共同调节者的后遗症1给敲入策略带来了一个重大问题。 因此，需要一种策略来筛选所有可能的点突变以确定功能上的重要 以及最有趣的修改。ES细胞是唯一自然永生的细胞系，同时忠实地 反映了它们在体内的作用，它们可以通过各种方式进行基因改造，以研究基因功能或 生成研究机理的模型系统。在这个项目中，我们建议分析一个 通过利用由蛋白质组学资源生成的PTM数据并通过利用 胚胎干细胞具有许多功能优势。为了验证ES细胞中系统的开发，我们 我们最初的“概念验证”工作将集中在SRC/p160基因家族上，然后是ES的构建 在其他协调控子中有翻译后改变的细胞。因此，我们将发展一种高吞吐量 在ES细胞中为每个辅活化子功能筛选大量的点突变。

It is our long-term goal to facilitate the identification of individual and combinatorial roles of PTMs of coregulators in differentiation, development, reproduction and metabolic homeostasis using the mouse as an eventual model. This coregulator 'postranseome1 presents a significant problem for knock-in strategies. Consequently, a strategy is needed to screen all possible point mutants to identify the functionally important and most interesting modifications. ES cells are the only cell lines that are naturally immortal, while faithfully reflecting their in vivo role, and they can be genetically modified in various ways to study gene function or generate model systems to study mechanism. In this project, we propose to analyze the function of a constellation of PTMs by leveraging PTM data generated by the Proteomics Resource, and by exploiting the many functional advantages available in ES cells. To validate the development of systems in ES cells we will focus our initial 'proof of concept' efforts on the SRC/p160 gene family, followed by construction of ES cells with post-translational alterations in other coregulators. Thus, we will develop a high throughput platform in ES cells to functionally screen a large number of point mutants for each coactivator.