Nanomedicine Center for Nucleoprotein Machines

核蛋白机器执行基本的生物过程，包括合成、修饰和修复 DNA和RNA。我们建议建立一个纳米医学开发中心(NDC)，专注于一个模型 执行DNA双链断裂的非同源末端连接(NHEJ)的纳米机器。这和 其他DNA修复机器具有相对简单的结构(20个组件)和重要的生物学意义 和临床相关性。DNA损伤修复对人类健康至关重要，因为两者都是正常代谢 活动和环境因素可能会导致DNA损伤，导致多达10万人 每天每个细胞的分子损伤。如果任其累积而不修复，这些损伤就会干扰基因 转录和复制，导致过早衰老、细胞凋亡或细胞分裂失控。我们有 组建了一个来自八个机构的跨学科团队，在细胞和分子方面拥有重要的专业知识 DNA损伤修复生物学、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 仪器仪表。我们将开发创新的纳米技术和生物分子方法来阐明 DNA修复纳米机器内部和之间的结构-功能关系。正在形成的一般原则 这些研究将为精确修改存储在DNA和RNA中的信息奠定基础， 最终导致针对包括癌症在内的一系列疾病的新的治疗策略。NDC 有五个密切相关的目标包括：(1)开发正交蛋白质标签策略和新颖的 包括用于纳米机器靶向的量子点生物偶联物的荧光探针；(2)解密 核心NHEJ反应所需组分的结构-功能关系；(3)表征 在修复焦点的背景下纳米机器组装和拆卸的动力学；(4)确定 固定单元中高分辨率修复灶的尺寸和结构；(5)制定工程设计 DNA双链断裂修复的基本原理。该NDC将补充现有的NDC，这些NDC专注于 关于细丝、膜和蛋白质折叠酶，以及开发的探针、工具和方法 将适用于广泛的生物学和疾病研究。我们的长期愿景是提供基因 基于操纵体细胞人类基因组的能力的常见人类疾病的治疗 纳米医学方法是廉价、有效和用户友好的，类似于今天的疫苗接种。

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 approaches that are inexpensive, effective, and user-friendly, similar to vaccination today.