Collaborative Research: Dynamic degradation of proteins by ubiquitination provides a novel therapeutic for controlling elevated protein levels

严格控制蛋白水平的能力对细胞健康至关重要。高水平会导致癌症、糖尿病、痴呆和心脏病等疾病。目前的治疗方法并不试图降解多余的蛋白质。我们建议开发一种触发蛋白质降解且可逆的技术。针对公众和教师的活动将扩大对科学、技术、工程和数学（STEM）培训重要性的理解。让高中生和本科生接触动手实验室研究经验将有助于吸引和留住STEM领域的学生。最终，这种培训将造就具有全球竞争力的劳动力。该项目将开发一种新的有效方法，通过合成偶联和引导天然蛋白酶体降解机制向所需的靶蛋白降解来诱导蛋白质降解。我们将重新利用Cas6家族蛋白作为位点特异性RNA结合和加工的通用平台，并展示它们在基于细胞内microRNAs （miRNAs）水平的蛋白质水平动态微调方面的实用性。创造一种能够适应不断变化的细胞环境并允许靶向必需蛋白质的新技术的关键挑战是实现蛋白质水平的动态微调，这在目前是不可能的。为此，我们将评估不同的策略来关闭蛋白质降解，使目标蛋白质可以维持在一个理想的或设定的水平。我们设想我们的技术广泛适用于任何蛋白质，因此将影响许多蛋白质水平升高导致或驱动其病理的疾病。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。

The ability to tightly control protein level is critical for cell health. High levels contribute to diseases that include cancer, diabetes, dementia, and heart disease. Current treatments do not attempt to degrade excess proteins. We propose to develop a technology that triggers protein degradation and that is reversible. Activities targeted to the general public and to teachers will expand the understanding of the importance of training in science, technology, engineering and mathematics (STEM). Exposing high school and undergraduate students to hands-on laboratory research experiences will help to attract and retain students in STEM fields. Ultimately, this training will result in a globally competitive workforce.This project will develop a new and potent approach that induces protein degradation by synthetically coupling and directing the native proteasomal degradation machinery towards the desired target protein(s). We will repurpose the Cas6 family proteins as a generalizable platform for site-specific RNA binding and processing, and demonstrate their utility to provide dynamic fine-tuning of protein levels based on intracellular microRNAs (miRNAs) levels. A key challenge in creating a novel technology that can adapt to the changing cellular environment and permit the targeting of essential proteins, which presently is not possible, is to enable dynamic fine-tuning of the protein levels. Towards this end, we will evaluate different strategies to turn off the protein degradation such that the target protein can be maintained at a desirable or set level. We envision that our technology is broadly applicable to any protein and thus will impact many diseases wherein elevated proteins levels contribute to or drive their pathology.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.