Exploit decades of mechanistic understandings to develop new routes to synthesis of RNA in high yield and purity Specifically, develop approaches targeting mRNA vaccines and therapeutics, CRISPR therapies and technologies, and advances in long noncoding RNAs, with a particular eye towards inexpensively eliminating the undesired innate immune response.
The standard enzymatic batch synthesis models of RNA production contribute to lower yields and purity than the enzyme is capable of producing. The Martin lab is developing new mechanism-based approaches toward RNA synthesis that will optimize the process, reducing costs and the time to synthesis.
Applied and basic science centered around RNA is expanding dramatically (and synergistically). In these endeavors, fidelity and purity of synthetic RNA are both essential, while faster turn around and lower overall cost will drive RNA technologies forward. Examples include mRNA vaccines, mRNA therapeutics and CRISPR therapeutics, but future applications will also include diseases of RNA splicing and the ever growing landscape of noncoding RNAs in biology.
We are developing several approaches towards improved enzymatic synthesis of RNA, at all length scales. Simple modifications to the enzyme, the DNA, or both drive promoter binding to new levels. This allows challenging the system with salt to eliminate product re-binding in high yield batch reactions (product re-binding leads to dsRNAs that trigger the innate immune response). Eliminating the conversion of correct length RNA to dsRNA improves both yield and purity.
We are developing a novel flow reactor to replace batch (and flow-batch) syntheses. This allows for the continuous removal of RNA from the reactor/enzyme, eliminating product re-binding and allowing immediate, real-time stabilization of the RNA.
To complement those efforts, we are developing simple purification approaches to select only correct length RNAs. We are also developing new analytical approaches to more fully characterize the purity of long RNAs.
