Synthetic modified RNA (modRNA) is a novel vector for gene transfer to the heart and other organs. modRNA can mediate
strong, transient protein expression with minimal induction of the innate immune response and risk for genome integration.
modRNA is already being used in several human clinical trials, and its use in basic and translational science is growing.
Due to the complexity of preparing modRNA and the high cost of its reagents, there is a need for an improved, cost-efficient
protocol to make modRNA. Here we show that changing the ratio between anti-reverse cap analog (ARCA) and
N1-methyl-pseudouridine (N1mJ), favoring ARCA over N1mJ, significantly increases the yield per reaction, improves
modRNA translation, and reduces its immunogenicity in vitro. This protocol will make modRNA preparation more accessible
and financially affordable for basic and translational research.
author: Yoav Hadas,Nishat Sultana, Elias Youssef, Mohammad Tofael Kabir Sharkar, Keerat Kaur, Elena Chepurko and Lior Zangi