Northwest Genome Engineering Consortium

这份U54申请描述了西北基因组工程联合会，一个调查小组 它将采用跨学科的方法来开发基因修复的概念、方法和工具， 治疗骨髓和淋巴系统单基因疾病的新方法。 尽管在传统治疗和第二代治疗方面取得了进展，但单基因疾病 淋巴造血在美国国内和全球范围内都对健康产生了重大影响。因此，这些 疾病是开发新的治疗方法的一个重要领域。基因组工程 技术已经进步到了目前可以修改单个缺陷基因的地步 原代人类细胞，这一过程被称为基因修复。奥维斯特基因组工程的目标 基因修复联盟(NGEC)是开发和应用基因修复治疗单基因白血病的组织 淋巴造血病。这一目标将通过两大科学目标来实现。第一 AIM将支持一种结合计算蛋白质设计、定向蛋白质进化、 结构分析，以及体外生化和分子分析，目标是开创 生产人工LAGLIDG归巢内切酶(LHEs)，用于各种类型的基因组 工程应用。第二个目标将支持团队方法来应用LHE‘s for Site 造血细胞中的特异性基因修复。这种方法将把在目标1中创作的小说LHE与 使用非整合慢病毒载体导入LHE和修复/修饰模板 以造血干细胞为目的进行基因修复。它还将探索新的操纵方法 内源性DMA修复机制增强LHE诱导的基因修复的原位效应 造血干细胞，并开发自体造血干细胞植入方法 针对基因修复应用进行了优化。除了专注于将基因修复应用于造血干细胞 细胞，NGEC将为基因组工程作为更广泛的科学学科的发展做出贡献 通过：1)博士后培训计划，为合格的研究员提供获得 在基于LHE的基因组工程过程中的经验和培训；以及2)关于 基因组工程，将向来自任何机构的学生和教职员工开放，并将包括演讲 由在基因组工程相关领域工作的知名外部调查人员提供。

This U54 application describes the Northwest Genome Engineering Consortium, an investigative team which will pursue an interdisciplinary approach to develop concepts, methods, and tools for gene repair, a new approach to the treatment of monogenic disorders of the bone marrow and lymphoid systems. Despite advances in conventional and 2nd generation therapies, monogenic disorders of lymphohematopoiesis have significant health impacts both within the US and on a global scale. Thus, these diseases are an important area for the development of new treatment approaches. Genome engineering technology has progressed to the point that it is presently feasible to modify individual defective genes in primary human cells, a process termed gene repair. The goal of the ^orthwest Genome Engineering Consortium (NGEC) is to develop and apply gene repair to the treatment of monogenic lymphohematopoietic disorders. This goal will be accomplished through two major scientific aims. The first aim will support a team approach that combines computational protein design, directed protein evolution, structural analysis, and in vitro biochemical and molecular analyses with the goal of pioneering methods for producing artificial LAGLIDADG homing endonucleases (LHEs), for use in a variety of types of genome engineering applications. The second aim will support a team approach to the application of LHE's for site specific gene repair in hematopoietic cells. This approach will combine novel LHE's created in Aim 1 with the use of non-integrating lentiviral vectors for the introduction of an LHE and a repair/modification template to hematopoietic stem cells for the purpose of gene repair. It will also explore new methods for manipulating endogenous DMA repair mechanisms to enhance the in situ efficacy of LHE-induced gene repair in hematopoietic stem cells, and develop methods for autologous hematopoietic stem cell engraftment optimized for gene repair applications. In addition to its focus on applying gene repair to hematopoietic stem cells, the NGEC will contribute to the development of genome engineering as a broader scientific discipline through: 1) a post-doctoral training program which will provide an opportunity for qualified fellows to obtain experience and training in the process of LHE-based genome engineering; and 2) an annual Workshop on Genome Engineering, which will be open to students and faculty from any institution, and will include talks by prominent outside investigators working in genome engineering related fields.