Synthesis of RNAs Containing Methylated Adenosine Residues and Their Use in Enhancing Translation

Background & Unmet Need

• Several diseases are associated with abnormalities in translation initiation or reduced protein production from specific mRNAs, including cancer and viral infection
• Gene therapies can also be limited by low levels of translation of therapeutic products
• Protein translation typically begins with the recruitment of the 43S ribosomal complex to the 5’ cap of mRNAs by a cap-binding complex
• Some transcripts are translated in a cap-independent manner through poorly understood mechanisms
• Unmet Need: Methods to enhance mRNA translation and protein production for basic research and therapeutic applications

Technology Overview

• The Technology: A method for enhancing mRNA translation by incorporating N6-methyladenosine (m6A) in the 5’ UTR
• The Discovery: m6A is a reversible base modification seen in the 5’ UTR of many eukaryotic mRNAs which functions as an alternative to the 5’ cap to stimulate mRNA translation
• m6A can be incorporated in this position by including an adenosine methylation motif in the 5’ UTR
• m6A can then bind eukaryotic initiation factor 3 (eIF3), which is sufficient to recruit the 43S complex to initiate translation in the absence of the cap-binding factor eIF4E
• PoC Data: In the absence of eIF4E, m6A is sufficient to initiate translation in uncapped mRNA (p<0.01)
• Induction of cellular stress via heat shock increases cellular expression of mRNAs containing m6A in the 5’UTR

Technology Applications

• Kits to enhance protein yields in in vitro protein synthesis reactions
• Increased protein production for in vivo applications, such as gene therapy
• Enhanced mRNA translation for the treatment of diseases like cancer or viral infection

Technology Advantages

• Translation is independent of the 5’ cap or cap-binding proteins, allowing use in conditions wherein cap-dependent translation is suppressed
• Translation requires only a single m6A or other form of methyladenosine to be present within the 5’UTR, rather than multiple nucleotides throughout the transcript