Contribution to the completion of Comprehensive Mouse Knockout Resource

描述（由申请人提供）：本项目的总体目标是通过建立一个公开可用的、全面收集的完全基于C57 BL/6的空源敲除胚胎干细胞系，进一步发挥小鼠作为人类疾病研究中强大而重要的工具的价值，从中可以开发出小鼠基因组中每个基因均含有空突变的小鼠文库。本申请建议建立一个国际学术机构联盟，由四个加拿大实验室（Nagy、Hicks、Ding和Rancourt博士）、一个德国实验室（Wurst博士）和密苏里州大学比较医学中心（Critser博士）组成。开发该联合体是为了解决NIH RFA（RFA-HG-05-007），用于产生小鼠突变体的综合资源，其中小鼠基因组中的每个基因都已被标记有高实用性报告系统的无效突变敲除。该联合体满足理想资源的标准，因为所有突变都将在统一的背景菌株C57 BL/6上进行，这是小鼠研究人员最广泛使用的菌株。这个整体提案的独特之处在于，联盟中的一个实验室（Nagy博士的实验室）已经建立了一个特殊的C57 BL/6胚胎干细胞系，具有非常高的种系传递（~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，以在胚胎干细胞中进行高通量基因突变。结合现有的经验和三个主要实验室的专业知识，使无效突变，结合纳吉实验室的高生殖系效率的C57 BL/6胚胎干细胞系和密苏里州的大学的专业知识，在创造小鼠和操作质量控制/保证计划的大型NIH资助的资源，将提供一个高效和成本效益的“团队”，以实现这一RFA的目标。虽然有几个联盟成员参与了类似的世界范围的努力，以提供一个完整的图书馆的鼠标零模仿，没有重叠，这些项目和这个KOMP倡议。事实上，该提案的一个主要优势是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“启动和运行”，使该联盟能够以非常及时和具有成本效益的方式实现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，我们将能够确保持续获得和应用最前沿的技术和科学;从而在KOMP的过程中提供持续的质量改进。这一方面将确保在完成KOMP核心目标的过程中，沿着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.