London Calling 2023: Using nanopore sequencing to understand the manufacture, delivery, and action of mRNA vaccines

The lifecycle of an mRNA vaccine begins with its manufacture and formulation into lipid nanoparticles. The mRNA is then delivered to patients, where it is taken up by recipient cells and translated into the encoded protein. This study uses Oxford Nanopore cDNA and direct RNA sequencing to analyze each step in the mRNA vaccine lifecycle. We first show that Oxford Nanopore sequencing can be used during mRNA manufacturing to monitor mRNA quality and integrity, and detect contaminating RNA species that can induce unwanted inflammatory responses. Then, we directly measure the impact of different degradation pathways on mRNA integrity during formulation and storage. Finally, we describe the uptake, expression, and final degradation of mRNA vaccines within cells. Together, this study uses Oxford Nanopore sequencing to trace the lifecycle of an mRNA vaccine, providing quality data throughout the manufacturing process and insight into its mode of action within the cell.