Development of novel nano-agents to target DNA replication forks and modulate cell cycle and activity

开发新型纳米制剂以靶向 DNA 复制叉并调节细胞周期和活性

• 批准号: 2265127
• 金额: --
• 依托单位: University of Birmingham
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2265127
• 关键词: Development; novel; nano; agents; target

New agents that bind to DNA Y-shaped junctions will be studied to further understand the basic cell and molecular biology of these agents in more detail specifically: (i) how they enter the cell and where (and how quickly) they localise, (ii) how they interact with key biomolecules in the cell such as genomic DNA and the downstream cellular effects of those interactions and (iii) the resultant properties in in vitro cellular systems.Newly modified junction binders (e.g. His-tagged, fluorescently labeled) will be used to image and study cellular interactions and develop novel imaging tools to directly study biologically important DNA transactions (e.g. DNA replication forks) inside of cells by measuring incorporation of fluorescent base analogues and also using isolated DNA under laminar sheer flow. This technique will allow us to analyze the impact of the agents on rates of replication fork progression, frequency of fork stalling or collapse, and also will provide insights into the cellular response to exposure (compensatory origin firing, effect on nearby replication structures). In parallel, we will develop binders that are amenable to cross-linking by 'click' chemistry to identify proteins bound to the DNA within the vicinity of the agent.

将研究结合DNA y形连接的新药物，以进一步详细地了解这些药物的基本细胞和分子生物学：(i)它们如何进入细胞以及它们定位的位置（和速度）；（ii）它们如何与细胞中的关键生物分子（如基因组DNA）相互作用以及这些相互作用的下游细胞效应；（iii）在体外细胞系统中产生的特性。新修饰的连接结合剂（例如his标记，荧光标记）将用于成像和研究细胞相互作用，并开发新的成像工具，通过测量荧光碱基类似物的掺入，以及在层流纯粹流动下使用分离的DNA，直接研究细胞内生物学上重要的DNA交易（例如DNA复制叉）。这项技术将使我们能够分析药物对复制叉进展速率的影响，叉失速或崩溃的频率，并且还将提供对暴露的细胞反应的见解（代偿起源激活，对附近复制结构的影响）。与此同时，我们将开发可通过“点击”化学进行交联的结合剂，以识别与药物附近DNA结合的蛋白质。