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

5-三磷酸核苷规模化合成新方法及其在基因合成中的应用

• 批准号: 2748427
• 金额: --
• 依托单位: Durham University
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2748427
• 关键词: New; methods; scaled; synthesis; nucleoside

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.

我们的目标是为合成基因生产提供负担得起的修饰dNTPs合成路线，这将支撑生物技术和生物医学科学。我们的发现也将适用于广泛应用于学术和工业研究settings.BackgroundSince亚磷酰胺方法的出现和“基因机器”核酸工具的可用性在生物科学，医学和工业，包括下一代测序和合成基因的“繁荣”几次革命。目前，缺乏快速获得大阵列的高保真合成DNA是有效地并行探索各种基因产物的限制。Evonetix的技术平台，包括一种新的，互补配对的工程酶与设计的dNTP之间开发的达勒姆和Evonetix，有望消除这些bottlenecks.Research Plan初步发现从达勒姆将通过这个程序开发，提供一个平台，dNTP的生产。具体来说，学生将致力于：1。学生将探索磷酸化和亚磷酸化试剂的反应性，并将这些发现应用于流程中。Flow Chemistry提供易于扩展的可控、可重复的混合程序。2.提供改进的纯化dNTPs学生将探索使用现成的预包装的基于色谱介质的dNTPs的有效纯化。这些将与控制降水战略一起进一步探讨。在Evonetix的酶技术平台中使用dNTP学生将获得在Evonetix使用他们的dNTP产品的“动手”生物学经验，在酶测定开发和将这些工具应用于芯片上DNA合成。他们在Evonetix的时间也将使他们获得宝贵的创业经验，同时接触工程，产品开发，自动化，计算机合成，QC验证过程和真正跨学科商业化战略的许多其他方面。