New methods for scaled synthesis of nucleoside 5'-triphosphates and their application in gene synthesis

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We aim to deliver affordable synthetic routes for modified dNTPs for synthetic gene production that will underpin biotechnology and biomedical sciences. Our findings will also be applicable across NTP and dNTP systems that are widely used in academic and industrial research settings.BackgroundSince the advent of the phosphoramidite approach and the availability of 'gene machines' nucleic acid tools have powered several revolutions within the biosciences, medicine and industry, including the Next Generation Sequencing and Synthetic Genes 'booms'. Currently, the lack of rapid access to large arrays of high fidelity, synthetic DNAs is a limitation to exploring varied gene products efficiently, in parallel. The Evonetix technology platforms, including the novel, complementary pairing of an engineered enzyme with designed dNTPs developed between Durham and Evonetix, promises to remove these bottlenecks.Research PlanPreliminary findings from Durham will be developed through this programme to provide a platform for dNTP production. Specifically, the student will work on:1. Delivery of improved synthesis technologies for dNTPsThe student will explore the reactivities of phosphorylating and phosphitylating reagents and apply these findings in flow. Flow Chemistry offers controlled, repeatable mixing procedures that are readily scalable. 2. Delivery of improved purifications of dNTPsThe student will explore the use of off-the-shelf pre-packed cartridge-based chromatography media for efficient purifications of dNTPs. These will be further explored alongside controlled precipitation strategies.3. Use of dNTPs in enzymatic technology platform at EvonetixThe student will gain 'hands-on' biological experience of using their dNTP products at Evonetix, in enzyme assay development and applying these tools to on-chip DNA synthesis. Their time in Evonetix will also allow them to gain invaluable experience of a start-up alongside exposure to engineering, product development, automation, in-silico synthesis, the QC validation process and many other aspects of a truly interdisciplinary commercialization strategy.