Nanomedicine Center for Nucleoprotein Machines

说明(应用程序摘要) 核蛋白机器执行基本的生物过程，包括DNA和RNA的合成、修饰和修复。我们建议建立一个纳米医学开发中心(NDC)，专注于一种执行DNA双链断裂的非同源末端连接(NHEJ)的模型纳米机器。这种DNA修复机器和其他DNA修复机器具有相对简单的结构(20个组件)和重要的生物学和临床相关性。DNA损伤修复对人类健康至关重要，因为正常的代谢活动和环境因素都会导致DNA损伤，导致每个细胞每天有多达10万个分子损伤。如果任其积累而不修复，这些损伤会干扰基因转录和复制，导致过早衰老、细胞凋亡或细胞分裂失控。我们组建了一个由八个机构组成的跨学科团队，在DNA损伤修复、蛋白质标记和靶向、纳米结构探针、冷冻电子显微镜、信号细胞成像、定量图像分析和计算生物学以及光学显微镜仪器方面拥有丰富的细胞和分子生物学专业知识。我们将开发创新的纳米技术和生物分子方法，以阐明DNA修复纳米机器内部和之间的结构-功能关系。从这些研究中得出的一般原则将为精确修改存储在DNA和RNA中的信息奠定基础，最终导致对包括癌症在内的各种疾病的新的治疗策略。NDC有五个密切相关的目标，包括：(1)开发正交蛋白质标记策略和新型荧光探针，包括用于纳米机器靶向的量子点生物偶联物；(2)破译核心NHEJ反应所需组件的结构-功能关系；(3)在修复中心的背景下表征纳米机器组装和拆解的动力学；(4)在固定细胞中以高分辨率确定修复中心的尺寸和结构；(5)建立DNA双链和断裂修复的工程设计原则。这个NDC将补充现有的专注于细丝、膜和蛋白质折叠酶的NDDC，开发的探针、工具和方法将适用于广泛的生物学和疾病研究。我们的长期愿景是，基于使用纳米医学操纵体细胞人类基因组的能力，为常见的人类疾病提供遗传治疗。

DESCRIPTION (application abstract) Nucleoprotein machines carry out essential biological processes including synthesis, modification, and repair of DNA and RNA. We propose to establish a nanomedicine development center (NDC) focusing on a model nanomachine that carries out nonhomologous end joining (NHEJ) of DNA double strand breaks. This and other DNA repair machines have relatively simple structures (< 20 components) and significant biological and clinical relevance. DNA damage repair is vitally important to human health, as both normal metabolic activities and environmental factors can cause DNA damage, resulting in as many as 100,000 individual molecular lesions per cell per day. If allowed to accumulate without repair, these lesions interfere with gene transcription and replication, leading to premature aging, apoptosis, or unregulated cell division. We have assembled an interdisciplinary team from eight institutions, with significant expertise in cell and molecular biology of DNA damage repair, protein tagging and targeting, nanostructured probes, cryo-electron microscopy, signal-cell imaging, quantitative image analysis and computational biology, and light microscopy instrumentation. We will develop innovative nanotechnologies and biomolecular approaches to elucidate the structure-function relationships within and among DNA repair nanomachines. General principles emerging from these studies will lay a foundation for precise modification of the information stored in DNA and RNA, leading ultimately to novel therapeutic strategies for a wide range of diseases, including cancer. The NDC has five closely related aims including: (1) to develop orthogonal protein tagging strategies and novel fluorescence probes including quantum dot bioconjugates for nanomachine targeting; (2) to decipher structure-function relationship of components required for the core NHEJ reaction; (3) to characterize the dynamics of nanomachine assembly and disassembly in the context of repair foci; (4) to determine the dimensions and structure of repair foci at high resolution in fixed cells; (5) to establish the engineering design principles underlying DNA double-strand break repair. This NDC will complement existing NDCs that focus on filaments, membranes and protein folding enzymes, and the probes, tools and methodologies developed will be applicable to a wide range of biological and disease studies. Our long-term vision is to provide genetic cures for common human diseases based on the ability to manipulate the somatic human genome using Nanomedicine.