Creating a Genetically Encodable Nano-Compass to Navigate in Molecular Networks

描述 摘要 了解嵌入生物化学和机械网络中的各种生物分子如何在多个长度和时间尺度上协调以产生协调的细胞行为，需要在其天然细胞环境中表征其生物化学和生物物理特性。然而，这是一个重大的挑战，我们目前对生物分子的天然行为的理解受到缺乏合适的技术来测量和操纵它们在活细胞中的动态特性的限制。例如，为了揭示因果关系并实现对活细胞中的信号转导网络的系统理解，必须开发一套大的分子工具，旨在专门干扰原位信号传导通路并定量测量细胞反应。此外，很少有人知道的力学性能和行为的分子参与力的产生，细胞力学的维护和mechanotransduction在活细胞中由于，在很大程度上，缺乏实验方法的细胞内测量和操纵的生物物理特性。我建议通过整合分子工程策略，化学和细胞生物学方法，以及结合单分子成像和磁镊的最先进的生物物理测量和操作，开发遗传编码磁性探针等分子工具。这项工作将确定新的战略，同时测量和操纵细胞内的分子事件，并将提供使能技术探测活细胞中的生物化学和生物物理过程。在分子水平上更好地理解生命过程将导致对治疗这些过程失调引起的疾病的新见解。

DESCRIPTION Abstract Understanding how various biomolecules embedded within biochemical and mechanical networks are coordinated over multiple length and time scales to produce orchestrated cell behaviors requires characterization of their biochemical and biophysical properties in their native cellular context. However this represents a major challenge and our current understanding of the native behaviors of biomolecules has been limited by the lack of suitable techniques to measure and manipulate their dynamic properties in living cells. For example, to reveal causal connections and achieve a systems understanding of the signal transduction networks in living cells, a large set of molecular tools, designed to specifically perturb signaling pathways in situ and to quantitatively measure the cellular response, must be developed. Furthermore, very little is known about the mechanical properties and behaviors of molecules involved in force generation, cellular mechanics maintenance and mechanotransduction in a living cell due, in large part, to the lack of experimental methodologies for intracellular measurement and manipulation of biophysical properties. I propose to develop such molecular tools as genetically encodable magnetic probes by integrating molecular engineering strategies, chemical and cell biological approaches, and state-of-the-art biophysical measurement and manipulation combining single-molecule imaging and magnetic tweezers. This work will identify new strategies for simultaneous measurement and manipulation of intracellular molecular events and will provide enabling technologies for probing biochemical and biophysical processes in living cells. A better understanding of life processes at the molecular level will lead to new insights in treating diseases that result from dysregulation of these processes.