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

合作研究：通过泛素化动态降解蛋白质为控制蛋白质水平升高提供了一种新的治疗方法

• 批准号: 1803008
• 负责人: Wilfred Chen
• 金额: $ 30万
• 依托单位: University of Delaware
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1803008&HistoricalAwards=false
• 关键词: Collaborative; Research; Dynamic; degradation; proteins

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.

严格控制蛋白质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、痴呆症和心脏病等疾病。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的技术，这种技术是可逆的。针对普通公众和教师的活动将扩大对科学、技术、工程和数学(STEM)培训重要性的认识。让高中生和本科生接触到动手的实验室研究经验，将有助于吸引和留住STEM领域的学生。这个项目将开发一种新的有效的方法，通过综合耦合和引导天然蛋白酶体降解机制来诱导蛋白质降解(S)。我们将改变Cas6家族蛋白质的用途，将其作为特定位点RNA结合和处理的通用平台，并展示它们在基于细胞内microRNAs(MiRNAs)水平提供蛋白质水平动态微调方面的有效性。创造一种能够适应不断变化的细胞环境并允许靶向关键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，我们将评估关闭蛋白质降解的不同策略，使目标蛋白质能够保持在理想或设定的水平。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。

项目成果

Engineering a Blue Light Inducible SpyTag System (BLISS)

• DOI: 10.1021/jacs.1c03198
• 发表时间: 2021-06-02
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Hartzell, Emily J.;Terr, Justin;Chen, Wilfred
• 通讯作者: Chen, Wilfred

Dynamic modulation of enzyme activity by synthetic CRISPR–Cas6 endonucleases

• DOI: 10.1038/s41589-022-01005-7
• 发表时间: 2022-04
• 期刊: Nature Chemical Biology
• 影响因子: 14.8
• 作者: Alexander A Mitkas;M. Valverde;Wilfred Chen

A microRNA-gated thgRNA platform for multiplexed activation of gene expression in mammalian cells

用于哺乳动物细胞基因表达多重激活的 microRNA 门控 thgRNA 平台

• DOI: 10.1039/d2cc01478e
• 发表时间: 2022
• 期刊: Chemical Communications
• 影响因子: 4.9
• 作者: Hunt, Victoria M.;Chen, Wilfred
• 通讯作者: Chen, Wilfred