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Learn how to optimize double-stranded RNA (dsRNA) removal in in vitro transcription (IVT) mRNA through this comprehensive webinar presented by Senior Scientist Johnathan Parker from CELLSCRIPT™. Explore the origins of dsRNA as a problematic byproduct of in vitro transcription that significantly impacts synthetic mRNA applications by suppressing protein translation and triggering unwanted innate immune responses. Discover how even trace amounts of dsRNA can compromise mRNA-based research and therapeutic applications, making effective removal strategies crucial for success. Examine multiple mitigation approaches including T7 polymerase variants, chromatography-based purification methods, and RNase III digestion techniques while understanding their respective strengths and limitations through presented data analysis. Understand how reducing dsRNA content not only decreases interferon signaling but also substantially improves overall translational yield of mRNA products. Gain insights into combining these strategies, particularly RNase III treatment with nucleoside modifications such as pseudouridine or N¹-methylpseudouridine, to produce high-quality, low-immunogenic mRNA suitable for therapeutic applications. Master the decision-making process for selecting appropriate removal methods based on consistency across production runs and required dsRNA tolerance levels in final products. Develop practical skills to dramatically increase mRNA translation success in cellular applications while virtually eliminating immunogenicity concerns for enhanced therapeutic outcomes.
