A Novel Approach to Eliminate Length Impurities for High-Quality mRNA Production

PROJECT SUMMARY
With the success of mRNA-based Covid vaccines, there is immense interest in leveraging mRNA technology to
develop therapeutics for previously untreatable diseases. However, therapeutic applications, such as protein
replacement therapy, cardiovascular regeneration, and cancer immunotherapy, require mRNA of exceptional
purity to ensure safety and efficacy. Current mRNA production methods often yield mRNA with impurities, such
as double-stranded RNA (dsRNA), long RNA (LRNA), and short truncated (stRNA), which can provoke immune
responses and compromise therapeutic outcomes.
Cisterna has developed an innovative platform capable of synthesizing high-quality, purified mRNA, devoid of
product-related contaminants, making it ideal for therapeutic use while ensuring consistency and reproducibility
across batches. In this Phase I SBIR, our goal is to demonstrate the efficacy of our technology in eliminating
dsRNA, LRNA, and stRNA compared to conventional methods. We will employ sensitive fluorescence-based
methods and assays to quantify and eliminate these impurities, enabling the production of highly pure therapeutic
mRNA. Furthermore, we will validate our technology by assessing its impact on immunogenic response and
protein expression in mice, aiming for minimal to no immune activation while maintaining or enhancing
therapeutic efficacy. Successful completion of this project will provide a validated production platform for
contaminant-free mRNA synthesis, paving the way for Phase II studies focused on target identification, construct
design, and scale-up manufacturing. The mRNA therapeutics market is poised for significant growth, with biotech
and pharmaceutical companies actively developing mRNA-based therapeutics. We aim to leverage potential
partnerships to position ourselves as a key player in this dynamic field.

Grant Number: 1R43TR005424-01
NIH Institute/Center: NIH
Principal Investigator: Hari Bhaskaran