Zilberzwige-Tal Shai, Levin Aviad, Ezra Assaf, Elia Uri, Finkelstein-Zuta Gal, Kreiser Topaz, Gershon Lihi, Goldsmith Meir, Kon Edo, Navon Yoav, Cohen Ofer, Knowles Tuomas Pj, Peer Dan, Gazit Ehud
The Shmunis School of Biomedicine and Cancer Research, Tel Aviv University, Tel Aviv, 6997801, Israel.
Yusuf Hamied Department of Chemistry, Centre for Misfolding Diseases, University of Cambridge, Cambridge, CB2 1EW, UK.
Small. 2025 Aug;21(32):e2500114. doi: 10.1002/smll.202500114. Epub 2025 Jun 23.
RNA-based therapeutics have revolutionized precision medicine due to their unprecedented potency, specificity, and adaptability. However, the inherent limited stability of RNA, including mRNA used in vaccines, is a major obstacle to the full realization of their potential. This instability, coupled with the centralized nature of vaccine production, currently limits the generation of RNA therapeutics at the point of care, which will otherwise fully harness the potential of these agents. Here, a microfluidic platform is presented for on-demand, personalized synthesis of modified mRNA stabilized by lipid nanoparticles. The design includes trapped biotinylated DNA, tagged T7 RNA polymerase, and a Tesla mixer, allowing the on-chip synthesis, purification, and encapsulation of mRNA in uniform lipid nanoparticles (LNPs), all conducted seamlessly on the same microfluidic device. This on-chip microfluidic synthesis approach is found to match standardized mRNA production yields, yet surpasses typical purification methods. Furthermore, as a proof-of-concept, the versatility and efficacy of the platform are demonstrated by generating diverse RNA sequences and structures, exhibiting functionality in human cell lines and mouse models. Moreover, an active SARS-CoV-2 vaccine is successfully engineered, highlighting the platform's potential for personalized vaccination strategies and offering a promising avenue for high throughput, decentralized vaccine delivery, reduced cold chain dependence, and even advancing current personalized medicine approaches through custom RNA therapeutics.
基于RNA的疗法因其前所未有的效力、特异性和适应性,给精准医学带来了变革。然而,RNA固有的稳定性有限,包括疫苗中使用的mRNA,这是充分发挥其潜力的主要障碍。这种不稳定性,再加上疫苗生产的集中性质,目前限制了在护理点生成RNA疗法,否则这些疗法将充分发挥这些药物的潜力。在此,我们展示了一个微流控平台,用于按需、个性化合成由脂质纳米颗粒稳定的修饰mRNA。该设计包括捕获的生物素化DNA、标记的T7 RNA聚合酶和特斯拉混合器,允许在芯片上合成、纯化mRNA,并将其封装在均匀的脂质纳米颗粒(LNP)中,所有这些都在同一微流控装置上无缝进行。发现这种芯片上的微流控合成方法可达到标准化mRNA的产量,但优于典型的纯化方法。此外,作为概念验证,通过生成多样的RNA序列和结构,在人类细胞系和小鼠模型中展示功能,证明了该平台的多功能性和有效性。此外,成功设计了一种活性SARS-CoV-2疫苗,突出了该平台在个性化疫苗策略方面的潜力,并为高通量、分散式疫苗递送、减少冷链依赖,甚至通过定制RNA疗法推进当前的个性化医学方法提供了一条有前景的途径。
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