Semi-artificial nanomachines for detection of DNA damage and apoptosis

多用途分子尺寸机器系统的开发将是未来技术的基石 在生物和医学方面。该项目的目标是开发第一个实用的纳米机器设计，通过 将分子马达蛋白的活性生物分子与人工工程组件相结合。 该结构将是一种分子大小的机器，属于一种新的半人工纳米设备， 这是我们最近推出的。它将通过共价对特定的DNA断裂进行超快标记 附连荧光标记。自组装后，机器将使用自己的材料进行组装 两个探测器单元。在存在DNase I型(带有5‘PO4)或DNase II的钝端DNA断裂时 类型(含5‘OH)，探测器单元将选择性地附着在它们的目标DNA末端并对它们进行标记 有红色或绿色的荧光团。所描述的结构将是第一个检测到两个 固定组织切片中的特定DNA断裂类型和超高速传感器检测DNA损伤和 活的非固定细胞在几秒钟内发生凋亡。 该项目的具体目标是：1.设计并测试一种具有超能力的半人工纳米机器。 快速检测溶解DNA中不同类型的DNA损伤。2.开发生物医学应用 新设计的纳米机器使用它作为纳米传感器来检测两种主要类型的 固定组织中的细胞凋亡，基于DNA酶I和II型DNA断裂的标记。3.发展生物医学 新型纳米机器作为超高速纳米传感器在检测环境中的应用 未固定的活细胞中的DNA损伤和凋亡。 在Aim1中，我们将使用在溶液中具有特定量DNA断裂的模型系统来测试选择性， 检测的特异性和速度。在目标2中，我们将把我们的新结构应用于固定组织切片，并将 开发其作为原位检测细胞凋亡的纳米传感器的应用。在目标3中，我们将开发一种 应用我们的构建物作为超快传感器检测活细胞的凋亡，而不需要他们的固定。 半人工纳米机器将展示一种新的仿生方法来设计超快分子-- 这将是一种独特的检测DNA损伤和凋亡的工具，在生物和 医学研究。

Development of versatile molecular-size machine systemswill be the cornerstone of future technologies in biology and medicine. The goal of this project is to develop the first practical nanomachine design by integrating an active biological molecule of a molecular motor protein with artificially engineered components. The construct will be a molecular-size machine belonging to a new class of semi-artificialnanodevices, which we recently introduced. It will perform super-fast labeling of specific DMAbreaks via covalent attachment of fluorescent markers. After self-assembly, the machine will use its own material to fabricate two detector units. In the presence of blunt-ended DNA breaks of DNase I type (bearing 5' PO4) or DNase II type (bearing 5' OH), the detector units will selectively attach to their target DNA ends and will label them with either red or green fluorophores. The described construct will be the first nanosensor detecting two types of specific DNA breaks in fixed tissue sections and a super-fast sensor detecting DNA damage and apoptosis in live non-fixed cells within seconds. Specific Aims of the project are: 1. To design and test a semi-artificial nanomachine capable of super- fast detection of different types of DNA damage in solubilized DNA. 2. To develop a biomedical application of the newly designed nanomachine employing it as a nanosensor for detection of two major types of apoptosis in fixed tissues, based on labeling of DNase I and II type DNA breaks. 3. To develop a biomedical application of the newly designed nanomachine employing it as a super-fast nanosensor for detection of DNA damage and apoptosis in non-fixed live cells. In Aim1 we will use model systems with specific amounts of DNA breaks in solution to test selectivity, specificity and speed of detection. In Aim 2 we will apply our new construct to fixed tissue sections and will develop its application as a nanosensor for in situ detection of apoptosis. In Aim 3 we will develop an application of our construct as a super-fast sensor detecting apoptosis in live cells without their fixation. The semi-artificial nanomachine will exemplify a new bionic approach to the design of super-fast molecular- size devices and will be a unique tool for detection of DNA damage and apoptosis useful in biological and medical research.