Optimizing Modified mRNA In Vitro Synthesis Protocol for Heart Gene Therapy

Yoav Hadas, Nishat Sultana, Elias Youssef, Mohammad Tofael Kabir Sharkar, Keerat Kaur, Elena Chepurko, Lior Zangi

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

Abstract

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 (N1mΨ), favoring ARCA over N1mΨ, 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.

Original languageEnglish
Pages (from-to)300-305
Number of pages6
JournalMolecular Therapy Methods and Clinical Development
Volume14
DOIs
StatePublished - 13 Sep 2019

Keywords

  • gene therapy
  • in vitro mRNA synthesis
  • modified mRNA

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