An enzyme cascade enables production of therapeutic oligonucleotides in a single operation

Therapeutic oligonucleotides have emerged as a powerful drug modality with the potential to treat a wide range of diseases; however, the rising number of therapies poses a manufacturing challenge. Existing synthetic methods use stepwise extension of sequences immobilized on solid supports and are limited by their scalability and sustainability. We report a biocatalytic approach to efficiently produce oligonucleotides in a single operation where polymerases and endonucleases work in synergy to amplify complementary sequences embedded within catalytic self-priming templates. This approach uses unprotected building blocks and aqueous conditions. We demonstrate the versatility of this methodology through the synthesis of clinically relevant oligonucleotide sequences containing diverse modifications.

Automated summary

Therapeutic oligonucleotides have become a powerful drug modality for treating diseases, but the increasing number of therapies presents challenges in manufacturing. This study presents a biocatalytic method that can efficiently produce oligonucleotides in a single operation using polymerases and endonucleases to amplify complementary sequences. The method uses unprotected building blocks and aqueous conditions, and demonstrates versatility in synthesizing clinically relevant oligonucleotide sequences with diverse modifications.