CAREER: Synthesis of Multiple Architectures of Decodable Biohybrid Polymer Libraries

非技术总结：聚合物在社会中无处不在——从塑料瓶到手机屏幕上的防指纹涂层，这些材料是全球技术、医疗和环境进步的关键组成部分。尽管这种聚合物无处不在，但目前许多聚合物的设计和分析只考虑到一种特定的应用。要确定一种现有的聚合物适合用于不同的应用，可能需要数年时间，尤其是在新应用所需的化学和物理性质未知的情况下。为了将聚合物材料与新的应用相匹配，该提案描述了一个合成平台，可以同时筛选数十万种材料——这比目前合成聚合物的策略和获得诺贝尔奖的定向进化等技术高出一个数量级。为了促进高通量聚合物筛选，DNA将被用作将多个聚合物块聚集在一起的主要材料。模块化的装配策略允许少量的合成片段产生大量的杂化嵌段共聚物库。DNA钉也被设计成携带识别信息，可以帮助解码哪些聚合物块被钉在一起，即使是在复杂的混合物中。一旦聚合物块被钉在一起，由此产生的混合材料就可以被筛选出所需的特性（比如保护蛋白质免受恶劣环境影响的能力，这对于稳定胰岛素等基于蛋白质的药物很有用），然后使用DNA测序策略进行分离和鉴定。这个项目需要跨学科的灵感和技术，这是在课堂上经常被忽视的科学成果的一个方面。为了解决这种脱节，我们提出了两个基于当前成功框架的教育项目：(1)BioInspiring——将生物学启发的科学与日常项目联系起来的现场和虚拟活动；(2)Sci-athonU——一个学生合作竞赛，提出针对全球挑战的跨学科研究建议。这些教育计划的目标是增加对STEM资源的访问，这些资源强调交流和协作是科学的关键要素。这项拟议的研究计划的长期目标是揭示具有复杂功能的聚合物的设计原则，以推进国家医疗，技术和环境目标，同时提供新的方法来增加历史上在STEM中代表性不足的群体的学生的动机和保留。技术概述：可解码的聚合物文库有可能彻底改变合成大分子的结构-功能关系鉴定。虽然蛋白质数据库（PDB）包含近50,000种不同蛋白质的结构信息，但在合成大分子的不同结构空间中，表征材料的数量明显减少，使得结构-功能关系的识别处于起步阶段。高通量实验对生物化学产生了革命性的影响，RNA序列揭示了蛋白质表达的实时定量和定向进化，产生了独特的蛋白质结构-功能关系。尽管这些强大的组合方法的影响，合成聚合物的高通量实验仍然很少。在目前的技术下，每种聚合物必须并行表征，因为目前还没有从混合物中分离出聚合物的快速识别策略。受到这一基本差距的启发，这项工作旨在使用DNA“钉”来编码整个嵌段共聚物的聚合物组成，并提供表征聚合物形态和蛋白质稳定性的原理证明高通量分析。提出了设计线性、刷状和星形聚合物形态的合成策略，并提出了测序方案来鉴定分离的聚合物。该提案的最终目标是提供一个平台，在单个实验中将聚合物表征的能力从数百个扩展到数千个，从而能够快速识别聚合物结构和高级功能之间的新关系。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。

Non-Technical Summary: Polymers are pervasive in society – from plastic bottles to fingerprint resistant coatings on cell-phone screens, these materials are a critical component of global technological, medical, and environmental advances. Despite this ubiquity, many polymers are currently designed and analyzed with only one specific application in mind. It can take years to identify an existing polymer that would be appropriate for use in a different application, especially if the chemical and physical properties required for the new application are unknown. To help match polymer materials to new applications, this proposal describes a synthetic platform that will enable hundreds of thousands of materials to be screened simultaneously – a scale orders of magnitude above current strategies for synthetic polymers and rivaling technologies like Nobel prize-winning directed evolution. To facilitate high throughput polymer screening, DNA will be used as a staple to bring together multiple polymer blocks. The modular assembly strategy allows for few synthetic pieces to lead to large libraries of hybrid block copolymers. The DNA staples have also been designed to carry identifying information that can help decode which polymer blocks have been stapled together, even in complicated mixtures. Once polymer blocks have been stapled together, the resulting hybrid materials can be screened for desirable properties (such as the ability to shield proteins from harsh environments, which is useful for stabilizing protein-based drugs like insulin), then isolated and identified using DNA sequencing strategies. This project requires interdisciplinary inspiration and techniques, which is an aspect of scientific achievements that is frequently overlooked in the classroom. To address this disconnect, two education programs building on current successful frameworks are proposed: (1) BioInspiring – on-site and virtual activities connecting biology-inspired science to every-day items and (2) Sci-athonU – a collaborative student competition to generate interdisciplinary research proposals that target global challenges. The objective of these educational programs is to increase access to STEM resources that highlight communication and collaboration as critical elements of science. This proposed research program has the long terms goals of revealing design principles for polymers with sophisticated functions to advance national medical, technological, and environmental goals, while providing novel approaches to increasing the motivation and retention of students from groups historically underrepresented in STEM. Technical Summary: Decodable polymer libraries have the potential to revolutionize the identification of structure-function relationships for synthetic macromolecules. While the protein data bank (PDB) contains structural information for almost 50,000 distinct proteins, there are significantly fewer characterized materials within the diverse structure space of synthetic macromolecules, leaving the identification of structure-function relationships in its infancy. High-throughput experimentation has had a revolutionary impact on biochemistry with RNA seq revealing real-time quantitation of protein expression and directed evolution yielding unique protein structure-function relationships. Despite the impact of these powerful combinatorial approaches, high-throughput experimentation with synthetic polymers remains rare. With current techniques, each polymer must be characterized in parallel, as there is currently no strategy for the rapid identification of polymers isolated from a mixture. Motivated by this fundamental gap, this work aims to use DNA “staples” that encode the polymer composition throughout a block copolymer and to provide proof-of-principle high-throughput assays characterizing polymer morphology and protein stabilization. Synthetic strategies are proposed for the design of linear, brush, and star-like polymer morphologies assembled with strategically designed architectures, and sequencing protocols are proposed to identify isolated polymers. The ultimate goal of this proposal is to provide a platform that expands the capabilities of polymer characterization from hundreds to thousands of polymers in a single experiment, enabling rapid identification of novel relationships between polymer structure and advanced functions.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.