FMSG: Bio: Microbial Foundry for Distributed Manufacturing of mRNA-Containing Biomaterials

该提案将为含有信使RNA或mRNA的生物材料开发新的微啤酒厂式分布式制造策略。迫切需要可靠地生产、储存和分配含mRNA的材料。例如，目前全球范围内的COVID-19 mRNA疫苗短缺不仅是由于冷链的需要，也是由于集中制造和分销。由于mRNA是脆弱的，它通常需要脂质纳米颗粒的保护，只能在超低温下保存。在这种情况下，DNA折纸，一种依赖于寡核苷酸序列互补性的自组装方法，可以开发成具有携带mRNA潜力的热稳定药物载体。在这个项目中，研究人员将使用酵母生产大量的mRNA和寡核苷酸DNA序列，具有准确性和可扩展性。由此产生的单链DNA将基于计算机设计自组装成折纸纳米结构，这将笼住天然制造的具有所有所需化学修饰的靶向mRNA。DNA折纸笼将允许mRNA的纯化和收获。DNA架构将被编程用于按需mRNA释放，并3D打印成水凝胶制剂，用于稳定储存和给药。拟议的研究还将得到教育和劳动力发展计划的补充。拟议的活动将开发实践和伴随的基于实践的学习内容，为生物制造部门的未来劳动力做好准备。该研究的中心目标是创建一个变革性的生物制造平台，在那里，诸如酿酒酵母或S。酿酒酵母被编程以在分散的位置产生DNA笼状mRNA生物材料所需的工业规模的定制设计的核酸。研究目标包括：（1）对微生物过程的机械理解，以建立mRNA和寡核苷酸DNA的可扩展铸造厂;（2）开发可编程环境响应DNA折纸笼的一般原理;（3）简化制造工艺，其中生物反应器和热循环仪与药物制剂的3D打印机集成;以及（4）制造的含mRNA材料的功效。这项研究的基础知识可能会改变两个关键行业，核酸合成和治疗性生物材料。这个未来制造奖是由分子和细胞生物科学支持的。这个奖项反映了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。

This proposal will develop new microbrewery-like distributed manufacturing strategies for biomaterials containing messenger RNA or mRNA. There is a critical need for reliable manufacturing, storage, and distribution of mRNA-containing materials. For example, the current worldwide shortage of mRNA vaccines for COVID-19 is caused not only by a need for a cold chain, but also by centralized manufacturing and distribution. Since mRNA is fragile, it often requires lipid nanoparticle protection and can only be preserved at ultralow temperatures. In this context, DNA origami, a self-assembly method that relies on oligonucleotide sequence complementarity, can be developed into thermally stable drug carriers with the potential to carry mRNA. In this project, the investigators will use yeast to produce large volumes of mRNA and oligo DNA sequences with accuracy and scalability. The resulting single-stranded DNA will self-assemble into origami nanostructures based on in silico designs, which will cage a targeted mRNA that is natively made with all needed chemical modifications. The DNA origami cage will allow for the purification and harvest of mRNA. The DNA architectures will be programmed for on-demand mRNA release and 3D printed into a hydrogel formulation for stable storage and administration. The proposed research will also be complemented by an educational and workforce development program. The proposed activities will develop hands-on and accompanied instruction-based learning contents to prepare the future workforce for biomanufacturing sectors. The central goal of the research is to create a transformative biomanufacturing platform, where organisms such as Saccharomyces cerevisiae or S. cerevisiae are programmed to produce industrial-scale, custom-designed nucleic acids needed for DNA-caged mRNA biomaterials at decentralized locations. The research objectives include: (1) a mechanistic understanding of microbial processes to establish a scalable foundry for mRNA and oligonucleotide DNA; (2) the development of general principles for programmable environmentally responsive DNA origami cages; (3) streamlined manufacturing processes where bioreactors and thermal cyclers are integrated with 3D printers for drug formulations; and (4) the efficacy of the manufactured mRNA-containing materials. The fundamental knowledge from the proposed study could transform two critical industries, nucleic acid synthesis and therapeutic biomaterials. This Future Manufacturing award was supported by Molecular and Cellular Biosciences.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.