SBIR Phase I: Development of a Novel Platform for Cost-Efficient mRNA Production in Yeast

The broader impact of this Small Business Innovation Research (SBIR) Phase I project is to develop a platform technology that manufactures high-quality messenger RNA (mRNA) at 1/10th the cost of current systems. In Vitro Transcription (IVT), the primary method of synthesizing mRNA for therapeutics and vaccines, encounters significant challenges in the form of expensive patented raw materials, complex purification processes, and supply chain shortages. There is an urgent need to fundamentally redesign mRNA production to accommodate the growing demand, enhance access to affordable, high-quality mRNA, and resolve supply chain issues. This project aims to innovate mRNA production by transforming yeast cells into efficient mRNA factories and using advanced chromatographic techniques for purification. The proposed platform could streamline mRNA manufacturing to significantly reduce costs and to broaden the scope, applicability and accessibility of mRNA. This innovation aims to provide pharmaceutical companies, biotechnology firms, and research institutions in academia, with affordable high-quality mRNA for vaccine development, therapeutics, and research purposes. This democratization of mRNA technology should accelerate innovation across different fields, shorten time-to-market for new treatments, and expand mRNA applications in emerging markets. Additionally, it may improve access for populations in low- and middle-income countries (LMICs), significantly advancing global health. The proposed project seeks to overcome high costs and inefficiencies associated with current IVT methods. This project introduces a novel approach to mRNA production by overexpressing a ribozyme-mRNA fusion in yeast, which is then immobilized and precisely cleaved on-column upon addition of a specific substrate that activates the ribozyme. This innovative method facilitates the efficient release and subsequent purification of the targeted mRNA directly from an RNA fusion construct expressed in yeast. Key technical objectives include demonstrating stable expression of the target RNA fusion in yeast, establishing a robust on-column purification system, and validating the purity and potency of the purified mRNA. Achieving these goals will validate the platform's feasibility and facilitate scaling of the technology to produce large quantities of mRNA, from grams to kilograms, at reduced costs, thereby revolutionizing mRNA production for diverse applications. 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.