Develop, Optimize and Commercialize UltraMarathonRT: A new enzyme for unbiased transcriptomic and epitranscriptomic characterization of RNAs regardless of length or composition.

Summary RNA is the central biomolecule of life, as it connects the sequence of DNA to the production of proteins, and serves as a structural building block for critical components within the cell. Despite the importance of detecting, quantifying and sequencing RNAs in order to exploit their roles in biology and medicine, the field of RNA research remains limited by the tools available for monitoring and manipulating RNA. An enzyme called reverse transcriptase (RT) is the most foundational tool for converting RNA into cDNA, enabling sensitive detection and RNA sequencing. However, existing, commercially available RTs have significant deficencies that limit our ability to fully exploit them as engines for analysis and sequencing. This problem has now been solved with the discovery of powerful new ultraprocessive RTs, MarathonRT (MRT) and ultraMarathonRT (uMRT). Discovered by the Pyle Lab at Yale University, MRT is an exceptionally processive RT that copies kilobase-length RNAs (such as the 30kb coronavirus genome) in a single pass without inhibition by secondary structures or repeats within an RNA template. Unlike other RTs, performance is maintained at ambient temperatures, which eliminates RNA degradation and enables reagent kits that can be deployed without refrigeration. For commercialization, the high-performance version, uMRT, will be developed, optimized and incorporated into a suite of commercial products for mass distribution. This SBIR grant is intended to fund the work necessary to create, develop, test and optimize these uMRT products for the most common RT-PCR, qRT-PCR and sequencing applications. This grant will translate an innovative scientific discovery into full-fledged commercial products that will significantly improve research and clinical practice, underscoring the economic power of superior enzymatic 'hardware.' We will produce a family of products that boosts the sensitivity of RT-PCR and RNA sequencing by employing a powerful new generation of reverse-transcriptase (RT) enzymes. History has shown that innovation typically follows the creation of more powerful scientific tools, such as the creation of the commercial biotechnology industry that followed the development of recombinant DNA technology in the 1970s, or the emerging field of gene editing therapeutics following the discovery of CRISPR enzymes. The introduction of uMRT products is a similar watershed moment as the scientific community will, for the first time, be able to read and detect variation within whole transcriptomes, revealing previously 'invisible' information that is critical for understanding all layers of biological function, from patients to single cells.