Double Stranded RNA Formation And How RNA Polymerase Choice Can Reduce It

Double-stranded RNA is one of those stubborn byproducts that can quietly inflate downstream purification costs and compromise mRNA product quality. In this presentation, Patrik Engström, Principal Scientist at Primrose Bio, breaks down the molecular mechanisms behind dsRNA formation during in vitro transcription, including loopback dsRNA generation and abortive transcript priming on full-length RNA. The data shows that T7 RNA polymerase consistently produces significant dsRNA across varying DNA template concentrations, while a class of non-T7 enzymes generates remarkably little.

Watch the full video to understand the tradeoffs involved and how enzyme choice could reshape your IVT workflow.