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：脱氧核糖核酸或DNA表现出独特的特性，其范围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，计算机和复杂折纸样结构的多功能工具。这个被称为DNA纳米技术的集体应用领域依赖于对DNA序列的精确控制来实现潜水功能，并对人类健康和工业产生了重大影响。 DNA纳米技术面临的一个重大挑战是需要扩大非典型应用的DNA产生，而不会变得过于过于过于过于成本，环境有害或过于麻烦。为了应对这一挑战，该项目利用制药公司使用的生物反应器来生产核酸疫苗在19009年大流行期间，以扩大DNA产生。通过采用微生物，类似于酵母如何将小麦转化为啤酒，这种具有成本效益的可扩展方法具有产生DNA的数量，比当前方法的产量大了数十万倍。在这个规模上，可以利用DNA超出其序列的独特特性来创建具有新颖特征的材料。与几乎任何其他分子不同，DNA的形状可以控制并编织成外来的形式，以增强其增强的特性，例如编织模式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为精确操作提供了新的机会，从而可以创建可调材料。这项涵盖生物反应器的跨学科应用的研究提供了两个围绕DNA纳米技术的教育机会。 “科学混搭”计划使高中生可以通过现场演示来展示令人兴奋的结果，融合两种科学，例如化学和生物学。此外，还向本科生和研究生提供了生物反应器训练营，为他们提供了经营生物反应器的必要技能，同时支持与Lonza的Portsmouth NH NH设施的科学家的互动，工业生物反应器的本地专家。总体而言，这项研究旨在将DNA纳米技术提升到更广泛的规模，并生成具有创新和受控的结构 - 概述的新材料。技术摘要：这项研究的目的是利用双型型DNA（DSDNA（dsdna）生成生成型DNA Hydrose dna Hydrogels dna Hydrogels。尽管有许多DNA水凝胶的例子，但与成本，可持续性和批量制备有关的挑战阻碍了这些材料在许多最终用途应用中的翻译。这项研究进展的基础的假设是，获得革兰氏尺度量的双链DNA（DSDNA）将提供新的途径，以获取利用DSDNA独特的聚合物特性的批量材料，并获得前所未有的控制和对其结构实质关系的洞察力。这项研究的主要目的是提高对DNA-Hydrecels的基本理解，并建立决定其结构性关系关系的设计原理。新的创新方法的开发可以在学术实验室环境中进行革兰氏尺度生产DNA合成子。然后，这些合成子用作建立块，通过共价，超分子和酶促方法产生散装DSDNA水凝胶。通过重新利用用于研究基因表达和DNA拓扑的策略，其目的是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的质粒（pDNA）的纯化和推导，以支持水凝胶材料的成本效益有效产生。随后，建立了这些材料在物理和共价连接的水凝胶中的结构特征与机械和化学特性之间的连接。最终，教育组成部分通过提供有关生物反应器和DNA水凝胶有效利用的全面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，便捷和可持续的方法来访问DSDNA水凝胶（ii）通过新的交联策略获得独特的体积属性的系统投资，以及（iii）（iii）使用Polymer网络拓扑的数量和相关性，并通过批准的机械授予来反映nsf的批准。优点和更广泛的影响审查标准。