Collaborative Research: MFB: Ultra-Fast Development of Portable Small Molecule Sensor-Actuators

在这个由生物技术分子基金会(MFB)倡议、化学系化学测量和成像计划以及化学、生物工程、环境和运输系统(CBET)部生物传感计划资助的项目中，科罗拉多大学博尔德分校化学和生物工程系的Tim Whitehead、加州大学河滨分校的Sean Cutler和Ian Wheeldon以及威斯康星医学院的Francis Peterson共同开发了使用工程分子电路随意控制生物系统的新方法。为了做到这一点，他们将进化和表征多种化学传感器，并推导出设计规则，使其他人能够在任何感兴趣的生物体中快速产生新的化学控制的生物反应。这个项目试图了解这是如何发生的，并定义特定蛋白质受体的化学感知的极限。在这样做的过程中，他们的工作将对生物技术的各个领域产生广泛影响，包括对细胞疗法的精确控制、非法药物或环境污染物的传感以及代谢产物的微生物生产。该项目位于化学生物学、有机化学、细胞与生化工程和结构生物学的交界处，将为从事这一科学的学生提供各级跨学科的培训。他们的外展和教育计划的主要目标是吸引年轻人投身STEM(科学、技术、工程和数学)职业，并改善化学生物和工业生物技术方面的培训。外展计划涉及多管齐下的努力，重点是大学预科和社区大学的参与，通过以发现为基础的实验室吸引本科生，并通过所有三个表现站点的本科生项目的多种研究经验。化学诱导二聚(CID)模块的发现和开发对化学生物学和生物技术具有变革意义。化学诱导的二聚化系统使用户能够构建配体控制的系统来调节细胞和生化功能，但受到相对较少的可用蛋白质模块和控制配体的限制。该项目寻求发展所需的基础知识，以了解如何对CID模块进行重新编程，以识别新的配体，并确定这种系统可以识别的化学空间的限制。为了做到这一点，研究小组将建立和筛选通过计算设计的PYR1(PYRABACTIN抗性1)突变文库。PYR1是一种植物激素受体，通过CID机制发挥作用，并具有可延展的配体结合口袋。该项目将进化出新的基于PYR1的传感器，由一系列不同的类药物小分子控制。然后，这些传感器的结构、信息学和生化特性将被用来为PYR1支架制定“识别规则”，并建立基于PYR1的传感器设计所需的基础知识。同时，还将开发一种基于酵母的快速系统，以加速基于PYR1的传感器的开发。总而言之，这些研究预计将导致对使用PYR1系统的传感器设计的详细了解，并为这种使能和可推广的传感器设计技术建立坚实的基础。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

In this project, funded by the Molecular Foundation of Biotechnology (MFB) initiative and the Chemical Measurement and Imaging program of the Division of Chemistry, and the Biosensing program of the Division of Chemical, Bioengineering, Environmental, and Transport Systems (CBET), Tim Whitehead of the Department of Chemical and Biological Engineering at the University of Colorado-Boulder, Sean Cutler and Ian Wheeldon at the University of California-Riverside, and Francis Peterson at Medical College of Wisconsin together are developing new ways to control biological systems at will using engineered molecular circuitry. To do this, they will evolve and characterize multiple chemical sensors and derive design rules that enable others to rapidly create new chemically controlled biological responses in any organism of interest. This project seeks to understand how this occurs and define the limits of chemical sensing by specific protein receptors. In doing so, their work will have broad impact on diverse areas of biotechnology, including the precise control of cellular therapies, sensing of illicit drugs or environmental contaminants, and the microbial production of metabolites. This project lies at the interface of chemical biology, organic chemistry, cell and biochemical engineering, and structural biology, and is therefore will provide cross-disciplinary training for the students engaged in this science at all levels. The primary goals of their outreach and education programs are to attract young people to careers in STEM (science, technology, engineering and mathematics) and improve training in chemical biology and industrial biotechnology. The outreach plan involves a multi-pronged effort focused on pre-college and community college engagement, engaging undergraduates through discovery-based labs, and through multiple Research Experiences for Undergraduates programs at all three performance sites.The discovery and exploitation of chemical-induced dimerization (CID) modules was transformative for chemical biology and biotechnology. Chemical-induced dimerization systems have enabled users to construct ligand-controlled systems to modulate cellular and biochemical function, but are constrained by a relatively small number of available protein modules and controlling ligands. This project seeks to develop the foundational knowledge required to understand how CID modules can be reprogrammed to recognize new ligands and to define the limits of chemical space that can be recognized by such systems. To accomplish this, the research team will build and screen computationally designed mutant libraries of PYR1 (PYRABACTIN RESISTANCE 1). PYR1 is a plant hormone receptor that functions through a CID mechanism and possesses a malleable ligand-binding pocket. The project will evolve new PYR1-based sensors controlled by a diverse range of drug-like small molecules. Structural, informatic, and biochemical characterization of these sensors will then be used to develop “rules of recognition” for the PYR1 scaffold and build foundational knowledge required for PYR1-based sensor design. In parallel, a rapid yeast-based system to accelerate PYR1-based sensor development will also be developed. Together, these studies are expected to lead to a detailed understanding of sensor design using the PYR1 system and establish a strong foundation for this enabling and generalizable sensor-design technology.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.