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

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.

Background
Since 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 Plan
Preliminary 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 dNTPs
The 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 dNTPs
The 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 Evonetix
The 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.

The CDT has five primary beneficiaries:
The CDT cohort
Our students will receive an innovative training experience making them highly employable and equipping them with the necessary knowledge and skillset in science and enterprise to become future innovators and leaders. The potential for careers in the field is substantial and students graduating from the CDT will be sought after by employers. The Life Sciences Industrial strategy states that nearly half of businesses cite a shortage of graduates as an issue in their ability to recruit talent. Collectively, the industrial partners directly involved in the co-creation of the proposal have identified recruitment needs over the next decade that already significantly exceed the output of the CDT cohort.
Life science industries
The cohort will make a vital contribution to the UK life sciences industry, filling the skills gap in this vital part of the economy and providing a talented workforce, able to instantly focus on industry relevant challenges. Through co-creation, industrial partners have shaped the training of future employees. Additional experience in management and entrepreneurship, as well as peer-to-peer activities and the beginning of a professional network provided by the cohort programme will enable graduates to become future leaders. Through direct involvement in the CDT and an ongoing programme of dissemination, stakeholders will benefit from the research and continue to contribute to its evolution. Instrument manufacturers will gain new applications for their technologies, pharmaceutical and biotech companies will gain new opportunities for drug discovery projects through new insight into disease and new methods and techniques.
Health and Society
Research outputs will ultimately benefit healthcare providers and patients in relevant areas, such as cancer, ageing and infection. Pathways to such impact are provided by involvement of industrial partners specialising in translational research and enabling networks such as the Northern Health Science Alliance, the First for Pharma group and the NHS, who will all be partners. Moreover, graduates of the CDT will provide future healthcare solutions throughout their careers in pharmaceuticals, biotechnology, contract research industries and academia.
UK economy
The cohort will contribute to growth in the life sciences industry, providing innovations that will be the vehicle for economic growth. Nationally, the Life Sciences Industrial Strategy Health Advanced Research Programme seeks to create two entirely new industries in the field over the next ten years. Regionally, medicines research is a central tenet of the Northern Powerhouse Strategy. The CDT will create new opportunities for the local life sciences sector, Inspiration for these new industries will come from researchers with an insight into both molecular and life sciences as evidenced by notable successes in the recent past. For example, the advent of Antibody Drug Conjugates and Proteolysis Targeting Chimeras arose from interdisciplinary research in this area, predominantly in the USA and have led to significant wealth and job creation. Providing a cohort of insightful, innovative and entrepreneurial scientists will help to ensure the UK remains at the forefront of future developments, in line with the aim of the Industrial Strategy of building a country confident, outward looking and fit for the future.
Institutions
Both host institutions will benefit hugely from hosting the CDT. The enhancement to the research culture provided by the presence of a diverse and international cohort of talented students will be beneficial to all researchers allied to the theme areas of the programme, who will also benefit from attending many of the scientific and networking events. The programme will further strengthen the existing scientific and cultural links between Newcastle and Durham and will provide a vehicle for new collaborative research.