ECA-PGR: Developing multiplexed transcriptional regulation systems for plant genome reprogramming

改变基因调控，也被称为转录重编程，是植物发育、繁殖和对各种胁迫反应的核心。同时对多个特定基因进行重编程是研究基因功能和表达、剖析复杂信号网络、工程代谢途径以及构建合成开关和电路所需要的。然而，允许生物学家以这种方式在植物细胞中开启和关闭多个基因的遗传工具目前还不可用。该项目开发了这些工具，不仅将有助于植物科学的基础研究，而且还将使植物科学家能够寻求新的解决方案，以应对诸如破坏性植物疾病、经济生物能源生产、可持续农业和应对气候变化等全球性挑战。本项目应用CRISPR植物基因组工程的最新技术进展，构建基本的综合性基因组重编程工具，并在水稻这一重要作物上进行演示。该项目的具体目标是开发1)一个高效的多路转录激活系统；2)高效的多路转录抑制系统；3)双向多路转录调控系统；4)集成基因组浏览器软件中的CRISPR设计工具，用于植物转录调控。本项目开发的高效植物转录重编程系统和设计工具将有利于植物研究界推进基础研究和作物改良。这些易于使用、有效的工具包将用于复杂的多基因、转录重编程测试，并提供给公共存储库Addgene的研究人员。此外，该项目将为不同层次的学生提供尖端培训，包括一名博士后科学家，两名博士生，以及许多来自不同背景的本科生和高中生。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Altering gene regulation, also known as transcriptional reprogramming, is at the heart of plant development, reproduction, and responses to various stresses. Simultaneously reprogramming multiple, specific genes is desirable for studying gene function and expression, dissecting complex signaling networks, engineering metabolic pathways, and building synthetic switches and circuits. However, genetic tools that would allow biologists to turn multiple genes on and off in this way in plant cells are currently unavailable. This project develops these tools and will not only aid basic research in plant science, but also allow plant scientists to seek novel solutions to global challenges such as devastating plant diseases, economical bioenergy production, sustainable agriculture and response to climate change. This project applies recent technical advances in CRISPR plant genome engineering to build essential comprehensive genome reprogramming tools and demonstrate them in rice, an important crop. The specific aims of this project are to develop 1) an efficient multiplexed transcriptional activation system; 2) an efficient multiplexed transcriptional repression system; 3) a bidirectional multiplexed transcriptional regulation system; and 4) a CRISPR design tool within the software of the Integrated Genome Browser for transcriptional regulation in plants. The highly efficient plant transcriptional reprogramming systems and the design tool developed in this project will benefit the plant research community in advancing basic research and crop improvement. These easy-to-use, effective tool packages will be tested for sophisticated multigene, transcriptional reprogramming and made available to researchers at the public repository, Addgene. In addition, the project will provide cutting-edge training to a diverse group of students at various levels, including a postdoctoral scientist, two PhD students, and many undergraduate and high school students from diverse backgrounds.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.