Metal nanocluster-modified nucleotides

金属纳米簇修饰的核苷酸

• 批准号: 323263560
• 负责人: Professor Dr. Andreas Marx
• 金额: --
• 依托单位: Department of Chemistry
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2020-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/323263560?language=en
• 关键词: Metal; nanocluster; modified; nucleotides

In the last two decades, metal nanoclusters (NCs) have made significant advances in the field of bioimaging, biosensing, therapeutic applications, and catalysis. Noble metal NCs like AgNCs and AuNCs, owing to their non-toxicity, biocompatibility, and photostability, have emerged as better luminescent materials over organic dyes and quantum dots. Thiol-capped AuNCs have been extensively studied in the field of catalysis, luminescence, and chemical sensing and exhibit very promising properties, calling for broad applications. The capability of DNA polymerases to accept dye-modified deoxynucleoside triphosphates (dNTPs) is exploited in many important biotechnological applications. These include next-generation sequencing approaches, single molecule sequencing, and labeling of DNA and PCR amplificates e.g. for microarray analysis. Furthermore, utilizing the intrinsic properties of DNA in combination with chemically introduced functionalities, provide an entry to new classes of nucleic acids-based hybrid material. Despite the impressive properties of metal NCs, these have, up to our knowledge, not yet exploited as modification for dNTPs and subsequently as substrate for DNA polymerases. In a collaborative approach will address this issue. We propose to design novel water-soluble nanoclusters modified with nucleotides in order to incorporate and label DNA by the action of DNA polymerases. We will be synthesize purines and pyrimidines with a suitably disposed thiol group used as ligand for obtaining the desired NC conjugates. After development of efficient synthetic routes, the NC-dNTP and NC-nucleoside conjugates will be spectroscopically characterized. In parallel, the NC-dNTP conjugates will be investigated towards their action on DNA polymerases with the aim to enzymatically incorporate the NC-modified nucleotides into the nascent DNA. If required, DNA polymerases will be engineered in order to improve the substrate properties of the NC-dNTP conjugates. By iterative rounds of synthesis and physical and biochemical studies, NC-dNTP-based systems will be obtained that have superior (photo)physical and biochemical properties than the systems available now and will allow broad applications.

在过去的二十年中，金属纳米团簇（NCs）在生物成像、生物传感、治疗应用和催化领域取得了重大进展。ag和AuNCs等贵金属NCs由于其无毒性、生物相容性和光稳定性，已成为有机染料和量子点之外更好的发光材料。巯基包盖AuNCs在催化、发光和化学传感等领域得到了广泛的研究，具有很好的应用前景。DNA聚合酶接受染料修饰的脱氧核苷三磷酸（dNTPs）的能力在许多重要的生物技术应用中得到了利用。这些包括下一代测序方法，单分子测序，以及DNA和PCR扩增物的标记，例如用于微阵列分析。此外，利用DNA的固有特性与化学引入的功能相结合，为新型核酸杂交材料提供了一个入口。尽管金属NCs具有令人印象深刻的特性，但据我们所知，它们尚未被用作dNTPs的修饰物，随后作为DNA聚合酶的底物。以协作的方式来解决这个问题。我们建议设计一种新的水溶性的核苷酸修饰纳米簇，以便通过DNA聚合酶的作用来整合和标记DNA。我们将合成嘌呤和嘧啶，以适当的巯基作为配体，以获得所需的NC偶联物。在开发了高效的合成路线后，将对NC-dNTP和nc -核苷缀合物进行光谱表征。同时，将研究NC-dNTP偶联物对DNA聚合酶的作用，目的是将nc修饰的核苷酸酶合到新生DNA中。如果需要，DNA聚合酶将被改造以改善NC-dNTP缀合物的底物性质。通过反复的合成以及物理和生物化学研究，将获得基于nc - dntp的系统，其具有比现有系统更优越的（光）物理和生物化学特性，并将允许广泛的应用。