Leveraging Next-Generation Directed Evolution Platforms and Chemical Control of Proteostasis to Deliver Robust Biotechnologies and Illuminate Roles of Chaperone Networks in Protein Evolution

利用下一代定向进化平台和蛋白质稳态的化学控制来提供强大的生物技术并阐明伴侣网络在蛋白质进化中的作用

• 批准号: 10610504
• 负责人: Matthew Donald Shoulders
• 金额: $ 6.73万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10610504
• 关键词: Address; Admission activity; Bacteria; Biological; Biology; Biotechnology; Cells; Chemicals; Complex; Country; Directed Molecular Evolution; Disease; Doctor of Philosophy; Ensure; Environment; Escherichia coli; Evolution; Goals; Grant; Human; Laboratories; Libraries; Malignant Neoplasms; Methodology; Molecular Chaperones; Mutation; Organism; Parents; Process; Proteins; Research; Role; System; Testing; Training; Tube; Variant; Yeasts; base; chemical genetics; experience; mutant; next generation; novel; programs; protein folding; proteostasis; success; technique development; tool

Directed evolution mimics and accelerates natural evolution in the laboratory in order to create useful new biomolecules and to study evolutionary processes. Although methodologies for directed evolution are well- established in test tubes and in simple organisms like Escherichia coli and yeast, there is still a major chal- lenge. Specifically, novel biomolecules derived from directed evolution campaigns in these platforms often fail to function when transferred to more complex cellular environments, such as that of human cells. To address this critical issue, our laboratory recently pioneered a directed evolution platform that can be used to repeatedly generate massive libraries of mutant biomolecules while continuously selecting and enriching the most func- tional variants directly in the human cell environment. From a chemical biology perspective, we are also deeply engaged in studying functions of the proteostasis network – a vital and unique aspect of the human cellular environment that ensures proteins are correctly folded, processed and trafficked. We have developed an array of chemical genetic tools to modulate proteostasis, and we are now primed to integrate these tools with our directed evolution platform to both evolve previously inaccessible biomolecule functions and gain a deeper un- derstanding of how cells solve protein folding problems. This supplement request seeks support to provide a post-baccalaureate training experience for Mr. Ra’Mal Harris, within the auspices of the parent R35 grant described above. The goal is to provide focused training and scientific exposure that will prepare Ra’Mal Harris for admission to and success at leading biomed- ical PhD programs around the country.

定向进化在实验室中模仿和加速自然进化， 新的生物分子和研究进化过程。尽管定向进化的方法学很好- 在试管和简单的有机体如大肠杆菌和酵母中建立，仍然有一个主要的挑战， lenge。具体而言，在这些平台中，从定向进化运动中衍生的新型生物分子通常失败， 当转移到更复杂的细胞环境中时，如人类细胞。解决 在这个关键问题上，我们的实验室最近开创了一个定向进化平台， 产生大量的突变生物分子库，同时不断选择和富集最具功能的， 在人类细胞环境中直接进行基因突变。从化学生物学的角度来看， 从事研究蛋白质稳态网络的功能-一个重要的和独特的方面，人类细胞 确保蛋白质正确折叠、加工和运输的环境。我们开发了一个阵列 化学遗传工具来调节蛋白质稳态，我们现在准备将这些工具与我们的 定向进化平台，既能进化以前无法实现的生物分子功能，又能获得更深层次的进化。 了解细胞如何解决蛋白质折叠问题。 这一补充请求寻求支持，以便为Mr. Ra'Mal Harris，在上述父母R35补助金的赞助下。目标是提供集中的 培训和科学接触，将准备拉马尔哈里斯录取和成功的领先生物医学- 全国各地的PhD课程。