Northwest Genome Engineering Consortium (Component 1 of 11)

这个U 54应用程序描述了西北基因组工程联盟，一个调查小组， 它将追求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。 尽管常规和第二代疗法取得了进展，但单基因疾病 淋巴造血在美国和全球范围内都具有显著的健康影响。因此这些 疾病是开发新治疗方法的一个重要领域。基因组工程 技术已经发展到这样的程度，即目前可以修改单个缺陷基因， 原代人类细胞，这一过程被称为基因修复。西北基因组工程的目标 NGEC是开发和应用基因修复治疗单基因 淋巴造血系统疾病这一目标将通过两个主要的科学目标来实现。第一 aim将支持一个团队的方法，结合计算蛋白质设计，定向蛋白质进化， 结构分析，体外生物化学和分子分析，目的是开拓方法， 产生人工LAGLIDADG归巢核酸内切酶（LHE），用于多种类型的基因组 工程应用第二个目标将支持团队方法，以应用现场LHE 造血细胞中的特异性基因修复。这种方法将联合收割机与目标1中创建的新型LHE相结合， 使用非整合慢病毒载体引入LHE和修复/修饰模板 用于基因修复的造血干细胞它还将探索新的操纵方法 内源性DMA修复机制，以增强LHE诱导的基因修复在原位的功效， 造血干细胞和开发用于自体造血干细胞移植方法 优化基因修复应用。除了专注于将基因修复应用于造血干细胞， 细胞，NGEC将有助于基因组工程作为一个更广泛的科学学科的发展 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，这将是开放给学生和教师从任何机构，并将包括会谈 在基因工程相关领域工作的杰出外部研究人员。

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.