This team selected 3'-O-allyl-rNTPs as choice of reversible terminators for their enzymatic RNA synthesis chemistry. It remains interesting to see if other well known 3'-O-blocking groups, such as 3'-O-azidomethyl and 3'-O-NH2 group, will be reported in the application of enzymatic RNA synthesis.
Scientists developed a new enzyme-based, template-independent RNA oligonucleotide synthesis technology (eRNA), offering a sustainable alternative to traditional chemical methods for manufacturing RNA therapeutics. RNA oligonucleotides have become increasingly important in treating diseases, with chemical synthesis methods facing challenges in scalability, atom economy, and process mass intensity. Scientists from Wyss Institute at Harvard University developed an aqueous-based enzymatic platform using mutant variants of CID1 poly(U) polymerase and reversible terminator nucleotides with a 3′-O-allyl ether blocking group. This platform enables writing natural and modified RNA oligonucleotides one base at a time without the need for a template sequence. This research achieved an average coupling efficiency of 95% and successfully demonstrated the synthesis of natural and modified RNA sequences up to ten cycles. The enzymatic process was also adapted to a solid-phase format, showing promise for large-scale production. Additionally, the researchers developed a method for removing the initiator sequence using endonuclease V, enhancing the purity of the final product. The enzymatic synthesis platform offers a more sustainable, efficient, and scalable method compared to traditional chemical synthesis, with the potential to meet the growing demand for RNA-based medicines while reducing environmental impact. Congratulations to all the contributors！ Daniel Wiegand Jonathan Rittichier Erkin Kuru George Church EnPlusOne Biosciences Reference: https://lnkd.in/eHRpKG5b #RNA #RNAtherapy #RNAtherapeutics #oligonucleotides #enzymaticsynthesis