Collaborative Research: Engineering MS2 Capsid Assembly Using Systematic Fitness Landscapes

合作研究：使用系统适应度景观设计 MS2 衣壳组装

• 批准号: 2043973
• 负责人: Danielle Tullman Ercek
• 金额: $ 50.04万
• 依托单位: Northwestern University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2043973&HistoricalAwards=false
• 关键词: Collaborative; Research; Engineering; MS2; Capsid; Collaborative; Research; Engineering; MS2; Capsid

Much of the biological world is made up building blocks, known as proteins, which can assemble into complex structures at scales not visible to the naked eye. The proposed work explores the self-assembly of a structure made of proteins called a virus-like particle. Understanding how the building blocks of viruses come together could be important for developing therapeutics and vaccines to fight them. The objective of this work is to uncover the rules for how this virus particle assembles and how it can be modified to improve its function without disrupting its structure. Outreach efforts will be pursued that include engagement and training of undergraduate and graduate students, and engagement with K-12 students and the general public via demonstrations in schools and at science festivals, discussions at local libraries and museums, and creation and release of an episode about this work on a top-ranked podcast series for children ages 6-12.Virus-like particles are an excellent model system to study protein self-assembly. These closed-shell protein containers tend to be composed of a single protein and can be expressed to high titers in bacteria. They also have the useful property of encapsulating their own genetic information. This proposal takes advantage of this property to study both the advantageous and deleterious effects of mutations on the self-assembly properties of virus-like particles using an efficient new directed evolution technique. Importantly, this technique uses a single step to generate libraries of the particles in which all sequence positions are systematically substituted with every natural amino acid. These libraries can then be exposed to various selection pressures, enabling identification of particles with desirable physical or biochemical properties via the encapsulated RNA in survivors of the selection. Using a variety of mutagenesis and functional selections, this work aims to achieve two specific objectives. First, it will elucidate the critical requirements for assembly of the virus-like particle derived from bacteriophage MS2. Second, it will improve the chemical reactivity and stability of the particle so it can remain intact following modifications for any desired application.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.

许多生物世界都是组成的，被称​​为蛋白质，可以在肉眼看不到的尺度上组装成复杂的结构。提出的工作探讨了由称为病毒颗粒的蛋白质制成的结构的自组装。了解病毒的基础如何聚集在一起对于开发治疗和疫苗来对抗它们可能很重要。这项工作的目的是揭示该病毒粒子如何组装的规则以及如何修改它以改善其功能而不会破坏其结构。 Outreach efforts will be pursued that include engagement and training of undergraduate and graduate students, and engagement with K-12 students and the general public via demonstrations in schools and at science festivals, discussions at local libraries and museums, and creation and release of an episode about this work on a top-ranked podcast series for children ages 6-12.Virus-like particles are an excellent model system to study protein self-assembly.这些封闭的蛋白容器倾向于由单个蛋白质组成，可以表达为细菌中的高滴度。他们还具有封装自己的遗传信息的有用属性。该建议利用该特性研究突变对病毒样颗粒自组装特性的有利和有害影响，使用有效的新定向进化技术。重要的是，该技术使用一个步骤来生成颗粒的库，其中所有序列位置都用每个天然氨基酸系统地取代。然后可以将这些文库暴露于各种选择压力上，从而通过选择的幸存者中的封装RNA鉴定具有理想的物理或生化特性的颗粒。这项工作使用多种诱变和功能选择，旨在实现两个特定的目标。首先，它将阐明源自噬菌体MS2的病毒样粒子组装的关键要求。其次，它将提高粒子的化学反应性和稳定性，以在对任何所需应用的修改后保持完整。该奖项反映了NSF的法定任务，并且使用基金会的智力优点和更广泛的影响审查标准，认为值得通过评估值得支持。