Maximizing Investigators' Research Award (R35 - Clinical Trial Optional)

项目概要： 进化上保守的Wnt信号通路在后生动物的发育过程中起着关键作用 和干细胞的维持。我的实验室的主要重点是了解 Wnt信号通路的生物学功能，通过破译其分子机制， 识别新的Wnt组件。我们的最终目标是利用这些信息来了解 Wnt途径的失调可导致人类中Wnt驱动的疾病。使用工具 生物化学，我的实验室在我们对Wnt的基本理解上取得了突破， 信号传导，包括第一个生化系统的发展， 纯化的蛋白质，和第一个数学模型（李-海因里希模型）的发展。 我们目前在实验室中使用便携式C-DiGit印迹扫描仪，该扫描仪速度慢，使用费力， 缺乏我们许多实验所需的灵敏度。此外， 我们的部门在一个单独的楼层，经常被大量使用，我们无法进入 这些文书及时。我们的仪器的局限性问题 实验室和缺乏部门的可访问性大大阻碍了我们的执行能力 我们的研究及时。因此，我们要求资金购买LICOR奥德赛 M成像系统，具有更高的吞吐量，灵敏度和动态范围， 克服这些障碍。此外，这台新机器将允许我们执行一项新技术： 细胞内蛋白质印迹，这将提高我们的方法的重现性和动态范围。 蛋白质测量

PROJECT SUMMARY: The evolutionarily conserved Wnt pathway plays a critical role during metazoan development and stem cell maintenance in the adult. The primary focus of my lab is to understand the biological function of the Wnt signaling pathway by deciphering its molecular mechanism and identifying new Wnt components. Our ultimate goal is to use this information to understand how deregulation of the Wnt pathway can lead to Wnt-driven diseases in humans. Using tools of biochemistry, my lab has made breakthroughs in our fundamental understanding of Wnt signaling, including the development of the first biochemical system, the reconstitution of active purified proteins, and the development of the first mathematical model (Lee-Heinrich model). We currently utilize a portable C-DiGit blot scanner in our lab that is slow, laborious to use, and lack the sensitivity required for many of our experiments. In addition, the imaging instruments in our department are on a separate floor and are often heavily used, and we are unable to access these instruments in a timely manner. The issues of the limitations of the instrumentation in our lab and the lack of departmental accessibility have significantly hampered our ability to perform our research in a timely manner. Thus, we are requesting funds to purchase a LICOR Odyssey M imaging system, which exhibits greater throughput, sensitivity, and dynamic range, to overcome these barriers. In addition, this new machine will allow us to perform a new technique: In-Cell Westerns, which will improve improved the reproducibility and dynamic range of our protein measurements.