Poster Session 4, 2:15 PM - 3:00 PM: Campus Center Auditorium [A28]

Presenter: Arundhati Bhat

Faculty Sponsor: Craig T. Martin

School: UMass Amherst

Research Area: Chemistry and Materials Science

ABSTRACT

mRNA therapeutics are an up and coming therapeutic platform with an expansive application in medicine. To facilitate the advancement of mRNA therapeutics the optimization of RNA manufacturing with limited impurities is essential. Currently there are several concerns in RNA manufacturing including cis primed extension, trans primed extension, 3′ overhang initiation and abortive RNAs. Such impurities can trigger an innate response hence impurities in mRNA should be limited. The development of the co-tethered approach minimizes dsRNA formation during in vitro transcription (IVT). In industry, NTP incorporation for IVT is limited. Through a design of experiments (DoE) approach, the goal is to maximize NTP incorporation to produce higher yields of RNA in combination with the co-tethered approach. dsRNA content will be assessed using a molecular beacon developed and designed to quantify the ratio of dsRNA to total RNA using fluorescence. The decided IVT conditions will be interpreted within a mechanistic framework to ensure consistency and reproducibility across different experimental conditions. By integrating these mechanistic insights with empirical DoE data, an optimized RNA manufacturing protocol can be established. This refined approach aims to increase the purity of mRNA produced by reducing immunogenic impurities and maximize NTP incorporation to drive higher RNA yields. Ultimately, such an approach broadens the scope of mRNA applications and furthers foundational mRNA research.