High Throughput Production and Cryopreservation of Knockout Mice

描述（由申请人提供）：国际敲除小鼠联盟（IKMC）的长期目标是开发哺乳动物基因组中每个蛋白质编码基因的小鼠靶向突变资源，研究界可以使用该资源阐明人类生物学和疾病中的基因功能。在这个框架内，KOMP项目第一阶段的目标是在五年内产生2500个特征良好的基因敲除（KO）系。该计划将在三个功能领域建立一个互动中心系统：KO菌株的生产和初步表型；广泛的表型出现;以及项目的信息学和数据库支持。本申请描述了我们在杰克逊实验室（JAX）建立KO小鼠生产和冷冻保存中心的计划。JAX拥有卓越的专业知识、经验和资源，能够高效、经济地大规模执行所需的中心功能。该中心将利用多种JAX资源，包括JAX资源库，一个主要的社区鼠标资源计划；生殖科学，在菌株冷冻保存和创新研究方面处于世界领先地位；以及出色的数据管理系统和专业知识。为了实现中心的目标，我们将：1)从IKMC ES细胞中产生多达2500个KO小鼠系；2)检测和分析KO小鼠的生存能力和生育能力，并检测各等位基因LacZ的表达模式；3)繁育、冷冻保存和分配动物和/或种质到KOMP的表型中心；4)通过使用创新技术降低向表型中心提供小鼠和种质资源的成本。我们将使用JAX现有的可扩展的鼠标数据管理和实验室信息管理系统（LIMS）来跟踪所有数据并将其传送到KOMP数据协调中心数据库（DCCDB）。与表型中心和DCCDB一起，该项目承诺为科学界提供关键资源。

DESCRIPTION (provided by applicant): The long-term goal of the International Knockout Mouse Consortium (IKMC) is to develop a resource of targeted mutations in mice for every protein-coding gene in the mammalian genome that the research community can use to elucidate gene function in human biology and disease. Within this framework, the objective of the first phase of the KOMP^ program is to generate 2,500 well-characterized knockout (KO) lines over five years. The program will establish a system of interactive centers in three functional areas: production and preliminary phenotyping of KO strains; broad-based phenotyping; and informatics and database support for the program. This application describes our plan to establish a KO Mouse Production and Cryopreservation Center at The Jackson Laboratory (JAX). JAX has the exceptional expertise, experience, and resources to efficiently and cost-effectively carry out the Center functions on the large scale required. The Center will leverage multiple JAX resources including the JAX Repository, a major community mouse resource program; Reproductive Sciences, a world leader in strain cryopreservation and innovative research; and outstanding data-management systems and expertise. To meet the Center objectives we will: 1) Generate up to 2,500 KO mouse lines from IKMC ES cells; 2) Test and analyze KO mice for viability and fertility, and examine LacZ expression patterns of each allele; 3) Breed, cryopreserve, and distribute animals and/or germplasm to KOMP^ Phenotyping Centers; and 4) Reduce the cost of providing mice and germplasm to the Phenotyping Centers through the use of innovative technologies. We will use existing, extensible mouse data-management and laboratory information management systems (LIMS) at JAX to track all data and communicate it to the KOMP^ Data Coordination Center Database (DCCDB). Together with the Phenotyping Centers and the DCCDB, this project promises to deliver a critical resource to the scientific community. RELEVANCE: The mouse is a critical animal model for understanding gene function and developing accurate models of human disease. This proposal aims to generate up to 2500 new mutant mouse lines as part of the KOMP Program, providing a valuable resource to the scientific community. The Jackson Laboratory has the outstanding expertise, experience, and resources to efficiently and cost-effectively meet this goal.