CAREER: Non-Equilibrium RNA/Protein Liquids and Intracellular Phase Transitions

在这个项目中，PI将重点阐述RNP液体的两个重要方面：1）它们从可溶性组分组装而成，以及2）它们的材料特性，这对它们的生物功能有很大影响。PI将首先采用自下而上的方法，利用纯化组分的体外系统研究活细胞的复杂性。这将通过利用先进的显微镜和图像分析技术以及新颖的分子动力学测量来补充。这些结合的体外/体内研究将使PI能够理解RNP液体的全部复杂性，并测试这些液滴的组装和性质由ATP依赖性生物活性调节的假设。这一建议的更广泛的影响包括通过在高中，本科和研究生阶段提供实验室研究培训来教育下一代生物药理学家;在本科阶段，高级论文研究的实验室培训将发挥关键作用。在课堂上，PI将开发关于定量生物学和软生命物质的新课程。这种教学是PI更广泛努力的一部分，旨在为普林斯顿新生的生物工程社区建立基础设施，并将其与校园内外的更大生物物理社区相结合。在K-12级别，PI与特伦顿公立学校建立了外展伙伴关系，重点是激励基尔默小学8年级学生中代表性不足的群体。PI将参观教室，进行关于软生命物质的实践演示，这些学生将参观PI的实验室。为了评估这一推广活动的效果，将使用基于科学相关态度测试（TOSRA）的定量评估工具，PI将与科学教育研究人员一起实施和分析。该项目由物理学系生命系统物理学项目和分子与细胞生物科学系细胞动力学与功能项目共同支持。

In this project, the PI will focus on elucidating two important aspects of RNP liquids: 1) Their assembly from soluble components, and 2) Their material properties, which strongly impact their biological function. The PI will study the complexity of living cells by first taking a bottom-up approach, utilizing an in vitro system of purified components. This will be complemented by utilizing advanced microscopy and image analysis techniques, and novel molecular dynamics measurements. These combined in vitro/in vivo studies will allow the PI to understand the full complexity of RNP liquids, and test the hypothesis that the assembly and properties of these droplets are tuned by ATP-dependent biological activity. The broader impacts of this proposal include educating the next generation of biophysicists by providing laboratory research training at the high school, undergraduate, and graduate levels; at the undergraduate level, laboratory training for senior thesis research will play a key role. In the classroom, the PI will develop new courses on Quantitative Biology and Soft Living Matter. This teaching is part of the PI's broader effort to build the infrastructure for growing Princeton's nascent Bioengineering community, and couple it to the greater Biophysics community on campus and beyond. At the K-12 level, the PI has developed an outreach partnership with the Trenton public schools, which focuses on inspiring under-represented groups of 8th grade students from Kilmer Elementary School. The PI will visit the classroom for hands-on presentations on Soft Living Matter, and these students will visit the PI's laboratory. To evaluate the efficacy of this outreach, quantitative evaluation tools based on the Test of Science Related Attitudes (TOSRA) will be used, which the PI will implement and analyze together with a science education researcher.This project is being jointly supported by the Physics of Living Systems program in the Division of Physics and the Cellular Dynamics and Function Program in the Division of Molecular and Cellular Biosciences.