Nanomedicine Center for Nucleoprotein Machines

核蛋白机器进行基本的生物过程，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）， 纳米机器进行DNA双链断裂的非同源末端连接（NHEJ）。这个和 其他DNA修复机器具有相对简单的结构（< 20个组件）和显著的生物学特性。 和临床相关性。DNA损伤修复对人类健康至关重要，因为正常的代谢和 活动和环境因素可导致DNA损伤，导致多达10万人 每个细胞的分子损伤如果任其积累而不修复，这些损伤会干扰基因表达。 转录和复制，导致过早衰老、凋亡或不受调节的细胞分裂。我们有 组建了一个来自八个机构的跨学科团队，在细胞和分子生物学方面具有重要的专业知识， 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，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DNA和RNA中存储的信息奠定基础， 最终导致了包括癌症在内的多种疾病的新的治疗策略。述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的 荧光探针，包括用于纳米机器靶向的量子点生物缀合物;（2）破译 核心NHEJ反应所需组分的结构-功能关系;（3）表征 修复焦点背景下纳米机器组装和拆卸的动力学;（4）确定 固定细胞高分辨率修复灶的尺寸和结构;（5）建立工程设计 DNA双链断裂修复的基本原理。该国家数据中心将补充现有的国家数据中心， 纤维，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遗传 治疗人类常见疾病的基础上，操纵人体基因组的能力， 纳米医学方法是廉价的，有效的，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.