CAREER: Sustainable DNA Hydrogel Production via Bioreactor-Derived Plasmid DNA

职业：通过生物反应器衍生的质粒 DNA 进行可持续 DNA 水凝胶生产

• 批准号: 2340569
• 负责人: Nathan Oldenhuis
• 金额: $ 70.78万
• 依托单位: University of New Hampshire
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-02-01 至 2029-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2340569&HistoricalAwards=false
• 关键词: CAREER; Sustainable; DNA; Hydrogel; Production

Non-Technical Description:Deoxyribonucleic acid, or DNA, exhibits unique properties extending beyond its central role as genetic information, making it a versatile tool for creating sensors, logic gates, computers, and intricate origami-like structures. This collective field of applications, known as DNA nanotechnology, relies on the precise control of DNA sequences to achieve diverse functions, holding significant implications for human health and industry. A substantial challenge facing DNA nanotechnology is the need to scale up DNA production for atypical applications without becoming cost-prohibitive, environmentally harmful, or overly cumbersome. To address this challenge, this project leverages bioreactors, used by pharmaceutical companies for producing nucleic acid vaccines during the COVID-19 pandemic, to scale up DNA production. By employing microorganisms, akin to how yeast transforms wheat into beer, this cost-effective and scalable approach has the potential to produce DNA in quantities that are hundreds of thousands of times greater than current methods yield. At this scale, the unique properties of DNA beyond its sequence can be harnessed to create materials with novel characteristics. Unlike almost any other molecule, DNA's shape can be controlled and woven into exotic forms to give it enhanced properties, like how a weave pattern can control the strength of a fabric. Furthermore, DNA's interactions with proteins and other molecules offer new opportunities for precise manipulation, allowing the creation of tunable materials. This research, which encompasses the interdisciplinary application of bioreactors, provides two educational opportunities centered around DNA nanotechnology. The 'Science Mash-up' program allows high schoolers to fuse two sciences, such as chemistry and biology, while showcasing the exciting results through live demonstrations. Additionally, a bioreactor boot camp is offered to both undergraduate and graduate students, equipping them with the necessary skills to operate a bioreactor while facilitating interactions with scientists from Lonza's Portsmouth NH facility, local experts in industrial bioreactors. Overall, this research aims to elevate DNA nanotechnology to a broader scale and generate new materials endowed with innovative and controllable structure-properties.Technical Summary:The objective of this research is to leverage inexpensive, scalable, and environmentally benign production of double-stranded DNA (dsDNA) from bioreactors to generate DNA hydrogels. Despite numerous examples of DNA hydrogels, challenges related to cost, sustainability, and bulk preparation hinder the translation of these materials in many end-use applications. The hypothesis underpinning this research advance is that access to gram-scale quantities of double-stranded DNA (dsDNA) will provide new paths to obtain bulk materials that utilize dsDNA’s unique polymeric properties and gain unprecedented control and insight into their structure-property relationships. The primary goal of this research is to advance the fundamental understanding of DNA-hydrogels and establish design principles that dictate their structure-property relationships. The development of new innovative methodologies enables gram-scale production of DNA synthons within academic laboratory settings. These synthons then serve as building blocks to create bulk dsDNA hydrogels through covalent, supramolecular, and enzymatic methods. By repurposing strategies used to study gene expression and DNA topology, the aim is to gain unprecedented control over hydrogel network topology and elucidate their fundamental properties. The initial research focuses on expanding the purification and derivation of plasmidDNA (pDNA) from bioreactors, facilitating the cost-effective and efficient production of hydrogel materials. Subsequently, connections between the structural characteristics and the mechanical and chemical properties of these materials in both physical and covalently linked hydrogels are established. Ultimately, the educational component capitalizes on the interdisciplinary nature of the research by offering comprehensive demonstrations and training focused on the effective utilization of bioreactors and DNA hydrogels. The expected innovations include: (I) The development of affordable, facile, and sustainable methods to access dsDNA hydrogels (II) The systematic investigation of unique bulk properties achieved through new cross-linking strategies and (III) The quantification and correlation of polymer network topology and entanglement with bulk mechanical properties.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.

非技术描述：脱氧核糖核酸（DNA）表现出独特的特性，超出了其作为遗传信息的核心作用，使其成为创建传感器、逻辑门、计算机和复杂的类似折纸结构的通用工具。 DNA 纳米技术依赖于 DNA 序列的精确控制来实现多种功能，对人类健康和工业具有重大影响。为了应对这一挑战，该项目利用制药公司在 COVID-19 大流行期间用于生产核酸疫苗的生物反应器，通过利用微生物来扩大 DNA 生产，类似于酵母小麦转化为酵母的方式。啤酒，这种经济高效且可扩展的方法有可能生产比现有方法产量高出数十万倍的 DNA，在这种规模下，可以利用 DNA 超出其序列的独特特性来制造材料。与几乎任何其他分子不同，DNA 的形状可以被控制并编织成奇异的形式，从而赋予其增强的特性，例如编织图案如何控制织物的强度此外，DNA 与蛋白质和其他分子的相互作用提供了新的机会。这项研究涵盖了生物反应器的跨学科应用，提供了两种以 DNA 纳米技术为中心的教育机会，“科学混搭”计划允许高中生融合两种科学，例如。化学和生物学，同时通过现场演示展示令人兴奋的结果此外，还为本科生和研究生提供生物反应器训练营，为他们提供操作生物反应器所需的知识，同时促进与龙沙新罕布什尔州朴茨茅斯工厂的科学家的互动和当地技能。总体而言，这项研究旨在将 DNA 纳米技术提升到更广泛的范围，并产生具有创新和可控结构特性的新材料。 技术摘要：这项研究的目的是利用尽管DNA水凝胶的例子很多，但与成本、可持续性和批量制备相关的挑战阻碍了这些材料在许多领域的转化。支持这一研究进展的假设是，获得克级数量的双链 DNA (dsDNA) 将为获得利用 dsDNA 独特的聚合特性和增益的散装材料提供新的途径。这项研究的主要目标是促进对 DNA-氢的基本理解，并建立决定其结构-性能关系的设计原则。新的创新方法的开发使得克级生产成为可能。然后，通过重新利用用于研究基因表达和 DNA 拓扑的策略，这些合成子可作为构建块，通过共价、超分子和酶促方法创建大量 dsDNA 水凝胶。获得对水凝胶网络拓扑的前所未有的控制并阐明其基本特性。最初的研究重点是扩大生物反应器中质粒 DNA (pDNA) 的纯化和衍生，促进水凝胶材料的成本效益和高效生产，随后建立结构之间的连接。最终，确定了这些材料在物理和共价连接水凝胶中的特性以及机械和化学性质，教育部分通过提供集中于有效的综合演示和培训来利用研究的跨学科性质。预期的创新包括：（I）开发负担得起的、简便且可持续的方法来获取 dsDNA 水凝胶（II）通过新的交联策略实现独特的整体特性的系统研究和（III）聚合物网络拓扑和纠缠与整体机械性能的量化和相关性。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力优点和更广泛的影响审查标准进行评估，被认为值得支持。