Novel Enzyme Specificities for Characterization of Large RNA Therapeutics

Advancements in large RNA therapeutics command the development of new technologies to characterize mRNA, sgRNA, and the associated impurities. Available enzymes and data analysis workflows are inadequate for rapid and sensitive characterization. Complementary cleavage specificities are needed to fill the sequence coverage gap generated by available RNases. Tunable dinucleotide specific enzymes generating shorter and longer digestion products and streamlined informatics enable accurate sequence confirmation with high efficiency and confidence to ensure integrity in a single LC-MS run.

Learning Objectives: 

1. Discuss the development of CRISPR sgRNA and mRNA therapeutics with the introduction of novel tools to lower the barrier for thorough characterization and monitor product quality.

2. Summarize the limited specificity of commercial enzymes for confident RNA sequence coverage and mapping. New RNases provide the ability to generate more unique products, provide full sequence coverage and modifications tied to potency and efficacy in a single LC-MS experiment.

3. Discuss comprehensive solutions to streamline and automate the confident confirmation of oligonucleotide sequence and robust oligo mapping workflows for convenient routine RNA analysis in a GMP compliant environment to support timely development of safe and efficacious nucleic acid therapeutics.