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。尽管这种普遍性，许多聚合物目前的设计和分析，只有一个特定的应用。可能需要数年时间才能确定适用于不同应用的现有聚合物，特别是在新应用所需的化学和物理特性未知的情况下。为了帮助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，该平台将使数十万种材料能够同时进行筛选-比目前的合成聚合物战略和诺贝尔奖获得者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嵌段聚集在一起的钉合物。模块化组装策略允许很少的合成片段产生大的混合嵌段共聚物库。DNA斯台普斯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。一旦聚合物嵌段被钉在一起，就可以筛选所得的杂交材料的所需特性（例如保护蛋白质免受恶劣环境影响的能力，这对于稳定基于蛋白质的药物如胰岛素是有用的），然后使用DNA测序策略分离和鉴定。这个项目需要跨学科的灵感和技术，这是科学成就的一个方面，在课堂上经常被忽视。为了解决这一脱节问题，提出了两个建立在当前成功框架基础上的教育计划：（1）生物学-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 -一个合作的学生竞赛，以产生针对全球挑战的跨学科研究提案。这些教育计划的目标是增加获得STEM资源的机会，这些资源强调沟通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提高学生的动机和保留率，这些学生来自历史上在STEM中代表性不足的群体。技术总结：可解码的聚合物库有可能彻底改变合成大分子的结构-功能关系的鉴定。虽然蛋白质数据库（PDB）包含了近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内，表征材料明显较少，使结构-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测序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，高通量的合成聚合物的实验仍然很少。使用当前技术，必须平行地表征每种聚合物，因为目前没有用于快速鉴定从混合物中分离的聚合物的策略。出于这一根本性的差距，这项工作的目的是使用DNA“斯台普斯”，编码整个嵌段共聚物的聚合物组合物，并提供证明的原则，高通量测定表征聚合物形态和蛋白质稳定。提出了合成策略的线性，刷状和星形聚合物形态的设计与战略设计的架构组装，并提出了测序协议，以确定孤立的聚合物。该提案的最终目标是提供一个平台，在一次实验中将聚合物表征的能力从数百种扩展到数千种，从而能够快速识别聚合物结构和高级功能之间的新型关系。该奖项反映了NSF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.