Fluorescent Molecular Oscillator for DNA Damage Detection

用于 DNA 损伤检测的荧光分子振荡器

• 批准号: 7500284
• 负责人: BenXiao Zhang
• 金额: $ 16.32万
• 依托单位: VIVID TECHNOLOGIES
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-26 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7500284
• 关键词: Apoptosis; Apoptotic; Biological; Categories; Cell Nucleus; Cells; Cellular biology; Class; Clinical Research; Color; Condition; DNA; DNA Damage; DNA Double Strand Break; DNA Fingerprinting; DNA Single Strand Break; Detection; Devices; Disease; Energy Transfer; Evaluation; Fluorescence; Fluorescence Resonance Energy Transfer; Generations; Hour; Label; Life; Measures; Methods; Molecular; Molecular Machines; Molecular Motors; Motor; Proteins; Range; Reaction; Reaction Time; Research Design; Sensitivity and Specificity; Signal Transduction; Suspension substance; Suspensions; Testing; Time; Topoisomerase; Type I DNA Topoisomerases; Vaccinia; base; cell fixing; design; drug development; human TOP1 protein; nanodevice; nanoimaging; nanosensors; nanosized; nanosystems; prognostic; research study; sensor

Fluorescent Molecular Oscillator for DNA Damage Detection In this proposal, we will design and test a new and useful nanodevice  a fluorescent molecular oscillator. We will develop an application for this device as an ultra-fast nanosensor for detection of DNA damage and apoptosis in cell suspensions. The sensor is the first representative of a class of semi-artificial molecular machines, a category of molecular devices which we recently introduced. In our nanodevice, the molecular motor protein continuously separates and religates two different fluorescently labeled DNA parts, which participate in fluorescence resonance energy transfer (FRET). When specific DNA breaks are present in cells, energy transfer between parts of the sensor is interrupted, leading to a fluorescence color change signaling detection of DNA damage and apoptosis. The detection reaction is very fast and takes less than a minute. Specific Aims of the project are: 1. To design and test this new type of molecular device  a light-emitting molecular oscillator, which will self-assemble from fluorescently labeled DNA and a molecular motor protein  vaccinia topoisomerase I. 2. To develop the first practical biomedical application of the newly created fluorescent molecular oscillator as a FRET-based nanosensor for the ultra-fast detection of specific DNA damage and apoptosis in live and fixed cells. In experiments we will determine sensitivity, specificity of the sensors and the time of reaction sufficient for the accurate detection of DNA breaks. The time of survival of molecular oscillators in reaction conditions will also be measured. Based on our results, we will modify the prototypical design and apply it for ultra-fast detection of apoptotic DNA breaks in suspensions of fixed and live cells. The nanosensor will be useful in theoretical and clinical research in apoptosis and drug development, and in evaluation of the effects of therapy in diseases in which the extent of apoptosis and DNA damage have prognostic value.

用于DNA损伤检测的荧光分子振荡器 在这个提案中，我们将设计和测试一个新的和有用的纳米器件  一 荧光分子振荡器我们将为该设备开发一个应用程序， 用于检测细胞悬液中DNA损伤和凋亡的超快纳米传感器。 该传感器是一类半人工分子机器的第一个代表， 这是我们最近推出的一种分子器件。在我们的纳米器件中， 分子马达蛋白连续分离和重新连接两个不同的荧光 标记的DNA部分，其参与荧光共振能量转移（FRET）。 当特定的DNA断裂存在于细胞中时，细胞各部分之间的能量转移， 传感器中断，导致DNA的荧光颜色变化信号检测 损伤和凋亡。检测反应非常快，不到一分钟。 该项目的具体目标是： 1.来设计和测试这种新型的分子装置  发光 分子振荡器，它将从荧光标记的DNA自组装， 一种分子马达蛋白  牛痘拓扑异构酶I 2.开发新创建的第一个实际生物医学应用程序， 荧光分子振荡器作为基于FRET的超快纳米传感器 检测活细胞和固定细胞中的特异性DNA损伤和凋亡。 在实验中，我们将确定传感器的灵敏度、特异性和检测时间。 反应足以准确检测DNA断裂。的存活时间 还将测量反应条件下的分子振荡器。根据我们的结果， 我们将修改原型设计并将其应用于凋亡的超快速检测， 固定细胞和活细胞悬浮液中的DNA断裂。 该纳米传感器将在细胞凋亡的理论和临床研究中有用， 药物开发，并在评估治疗的疾病，其中， 细胞凋亡和DNA损伤程度具有预后价值。