Aptamer biosensors to measure PAR dynamics in living cells

用于测量活细胞中 PAR 动态的适体生物传感器

• 批准号: 2282164
• 金额: --
• 依托单位: University of Manchester
• 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-2282164
• 关键词: Aptamer; biosensors; measure; PAR; dynamics

PolyADP-ribosylation (PARylation) is a reversible post-translational modification where poly ADP-ribose (PAR) chains are assembled onto acceptor proteins. PARylation regulates various fundamental cellular processes including chromatin organisation, DNA repair, transcription and replication. PAR chains are synthesised by PAR polymerases (PARPs), and catabolised by PAR glycohydrolases (PARGs); so PAR dynamics is carefully regulated by the balance between both enzyme classes. However, we currently lack the ability to measure PAR dynamics in living cells. The aim of the project is to explore fluorescent or other chemically modified and measurable aptamers as 'PAR biosensors' capable of binding to PAR chains in living cells; and to generate new tools to study PARylation and its role in DNA damage, repair and replication in normal cellular physiology and pathological states such as ovarian and prostate cancers, with the industrial partner, Aptamer Group. The student will generate synthetic PARylated peptides; create aptamer beacons and evaluate the PAR biosensor(s) in living cells. The student will receive a broad interdisciplinary skills training in state-of-the-art aptamer technology for developing novel biosensors, in vitro analytical techniques and cell biology methods (cell culture, time-lapse and immunofluorescence microscopy), and functional assays (e.g. apoptosis, replication stress). They will also gain a strong intellectual training in the relevant background (cell cycle, DNA replication and cancer), and in experimental design and strategic multidisciplinary approaches.

聚腺苷二磷酸核糖基化(PolyADP-核糖化)是一种可逆的翻译后修饰，聚腺苷二磷酸核糖(PAR)链组装在受体蛋白上。PAR化调控各种基本的细胞过程，包括染色质组织、DNA修复、转录和复制。PAR链由PAR聚合酶(PARP)合成，并由PAR糖水解酶(PARGs)分解代谢，因此PAR的动态受到两类酶之间的平衡的仔细调节。然而，我们目前缺乏在活细胞中测量PAR动态的能力。该项目的目的是探索荧光或其他化学修饰和可测量的适体作为能够与活细胞中的PAR链结合的‘PAR生物传感器’；并与工业合作伙伴适配集团合作，开发新的工具来研究PAR化及其在正常细胞生理和病理状态(如卵巢癌和前列腺癌)中的DNA损伤、修复和复制中的作用。该学生将产生合成的PAR化多肽；创建适配子信标，并在活细胞中评估PAR生物传感器(S)。学生将接受广泛的跨学科技能培训，内容包括用于开发新型生物传感器的最先进的适体技术、体外分析技术和细胞生物学方法(细胞培养、延时和免疫荧光显微镜)以及功能分析(例如细胞凋亡、复制压力)。他们还将在相关背景(细胞周期、DNA复制和癌症)以及实验设计和战略性多学科方法方面获得强有力的智力培训。