Contribution to the completion of Comprehensive Mouse Knockout Resource

描述（由申请人提供）：该项目的总体目标是通过建立一个公开的、全面的、完全基于C57BL/6的零资源敲除胚胎干细胞系，进一步提高小鼠作为人类疾病研究中一个强大而重要的工具的价值，从中可以开发一个包含小鼠基因组中每个基因的零突变的小鼠文库。本申请建议建立一个由四位加拿大博士组成的国际学术机构联盟。Nagy, Hicks, Ding and Rancourt)，一个德国实验室（Wurst博士）和密苏里大学比较医学中心（Critser博士）。该联盟的建立是为了解决NIH RFA （RFA- hg -05-007）对小鼠突变体的综合资源的生产，其中小鼠基因组中的每个基因都被高效用报告系统标记的零突变敲除。该联盟符合理想资源的标准，因为所有突变都将携带在一个统一的背景菌株C57BL/6上，这是小鼠研究人员最广泛使用的菌株。这一整体建议的独特之处在于，该联盟的一个实验室（Nagy博士的实验室）已经建立了一种特殊的C57BL/6胚胎干细胞系，具有非常高的种系传输率（~70%）。重要的是，该联盟的成员也是加拿大资助项目（NORCOMM; Nagy, Hicks， Ding和Rancourt实验室）和欧盟资助项目（EUCOMM； Wurst实验室）的一部分，该项目旨在对胚胎干细胞进行高通量基因突变。结合三个主要实验室的现有经验和专业知识，制造零突变，结合Nagy实验室的高生殖系高效C57BL/6胚胎干细胞系和密苏里大学在创建小鼠和运行质量控制/保证项目方面的专业知识，为nih资助的大型资源；将提供一个高效且成本有效的“团队”来实现该RFA的目标。虽然一些联盟成员参与了类似的全球努力，以提供完整的小鼠null限制库，但这些项目与KOMP计划之间没有重叠。事实上，这项建议的主要优势在于这些实验室的丰富经验和现有基础设施。现有的高通量、低成本的方法在我们的集体实验室中“启动并运行”，使该联盟能够以极其及时和具有成本效益的方式完成KOMP的目标。重要的是，由于该联盟由学术团体组成，其中有多个单独资助的、正在进行的与KOMP直接相关的研究项目，我们将能够确保不断获得和应用最尖端的技术和科学；从而在本KOMP的过程中提供持续的质量改进。这方面将确保在完成KOMP核心目标的过程中，每一步都采用最具成本效益的方法。

DESCRIPTION (provided by applicant): The overall objective of this project is to further the value of the mouse as a powerful and important tool in the study of human disease through the establishment of a publicly available, comprehensive collection of a fully C57BL/6-based null resource knockout embryonic stem cell lines, from which a library of mice containing a null mutation in every gene in the mouse genome can be developed. This application proposes to establish an international consortium of academic institutions comprised of four Canadian (Drs. Nagy, Hicks, Ding and Rancourt), one German (Dr. Wurst) laboratories and the University of Missouri Comparative Medicine Center (Dr. Critser). This consortium was developed to address the NIH RFA (RFA-HG-05-007) for the production of a comprehensive resource of mouse mutants in which every gene in the mouse genome has been knocked out by a null mutation marked with a reporter system of high utility. This consortium fulfills the criteria for an ideal resource in that all of the mutations will be carried on a uniform background strain, C57BL/6, which is the strain most widely utilized by mouse researchers. The unique aspect of this overall proposal is that one laboratory in the consortium (Dr. Nagy's laboratory) has established an exceptional C57BL/6 embryonic stem cell line with a very high germline transmission (~70%). Importantly, this consortium has members which are also part of a Canadian funded project (NORCOMM; Nagy, Hicks, Ding and Rancourt labs) and a European Union funded project (EUCOMM; Wurst lab) to perform high throughput gene mutations in embryonic stems cells. Combining the existing experience and expertise of three major laboratories making null mutations, in combination with the Nagy lab's high germline efficient C57BL/6 embryonic stem cell line and the University of Missouri's expertise in creation of mice and operation of quality control/assurance programs for large NIH-funded resources; will provide a highly efficient and cost effective "team" to achieve the goals of this RFA. While several consortium members are involved with similar world-wide efforts to provide a complete library of mouse null imitations, there is no overlap between those projects and this KOMP initiative. In fact, a major strength of this proposal is the vast experience and existing infrastructure in those laboratories. The existing high throughput, low cost methods that are "up and running" in our collective laboratories enable this consortium to accomplish the goals of the KOMP in an extremely timely and cost efficient manner. Importantly, because this consortium is comprised of academic groups, in which there are multiple, separately funded, ongoing research projects directly related to the KOMP, we will be able to ensure continuous access to, and application of, the most cutting-edge technologies and science; thereby providing continuous quality improvement over the course of this KOMP. This aspect will ensure the most cost effective methods are applied at each step along the way to completing the core goals of the KOMP.