Knock-in of Posttranslational Mutations of Nuclear Receptor Coregulator Genes

这是我们的长期目标，以促进识别的个人和组合作用的PTM的共调节 在分化、发育、生殖和代谢稳态中的作用， 最终模型这种共调节子的后转基因组1为敲入策略提出了一个重要的问题。 因此，需要一种策略来筛选所有可能的点突变体以鉴定功能上重要的突变体。 最有趣的修改。ES细胞是唯一的细胞系，是自然不朽的，而忠实地 反映了它们在体内的作用，并且它们可以以各种方式进行遗传修饰以研究基因功能， 生成模型系统研究机理。在这个项目中，我们建议分析一个 通过利用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 我们将把我们最初的“概念验证”工作集中在SRC/p160基因家族上，然后构建ES 其他辅助调节因子发生翻译后改变的细胞。因此，我们将开发高通量 在ES细胞中的一个平台，以功能性地筛选每个共激活因子的大量点突变体。

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.