Synthetic regulatory sequence designs for mRNA vaccines

Abstract

mRNA-based therapeutics have demonstrated significant potential for enhancing human health across various applications. Notably, mRNA vaccines, a prominent subset of this therapeutic approach, showcased their efficacy during the COVID-19 pandemic by safeguarding billions of lives. Despite their success, the full scope of mRNA vaccines in addressing diverse health concerns, such as cancer, remains constrained by existing limitations in tunability and targetability. A deeper exploration of mRNA vaccine regulation is inevitable to harness their complete capabilities. This thesis centers on comprehending and manipulating two pivotal regulatory domains within the mRNA molecule itself: the coding sequence and the 5’ untranslated region (UTR). Regarding the coding sequence, we engineered an mRNA vaccine candidate featuring a combined antigen coding region for SARS-CoV-2 to elicit a dual immune response against the virus. Our findings underscore that the resultant antigen exhibited interactions with distinct antibodies generated throughout the natural course of infection. This interaction profile potentially signifies a dual immune activity for enduring protection. Therefore, we practiced the essential stages of mRNA molecule manipulation requisite for an effective vaccine candidate. In parallel, we devised an innovative methodology for constructing synthetic 5’ UTR libraries tailored for selective expression within cancer cells. Collectively, this thesis advances our grasp of mRNA vaccine regulation and design. Considering the needs of the current state of mRNA vaccines, this heightened control over mRNA molecules promises novel avenues for addressing a spectrum of diseases.