Therapeutic Oligonucleotides are chemically modified strands of DNA that can interact with mRNA through Watson-Crick base pairing. Through chemical modifications at key positions, oligonucleotides gain enhanced binding affinity, lower cytotoxicity and nuclease resistance. However, their precise sequence and the number of necessary modifications make them a challenging target for synthesis that is currently answered by highly wasteful, and unscalable phosphoramidite chemistry. This work demonstrates the application of thermophilic Endonuclease Vs in an enzymatic oligonucleotide synthesis platform to produce oligonucleotides. The results show that EndoVs are active toward oligonucleotides with 2â sugar modifications and phosphorothiate backbone modifications. Moreover, Endonuclease V have been used together with a polymerase in the production of oligonucleotides from dNTP stocks. Overall, Endonuclease Vs are a promising family of enzymes that after further research and engineering can become a key tool in future oligonucleotide synthesis.
|Date of Award
|31 Dec 2022
- The University of Manchester
|Anthony Green (Supervisor) & Sarah Lovelock (Supervisor)