High Throughput Production and Cryopreservation of Knockout Mice

基因敲除小鼠的高通量生产和冷冻保存

• 批准号: 8876984
• 负责人: Stephen A Murray
• 金额: $ 64.94万
• 依托单位: JACKSON LABORATORY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-07 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8876984
• 关键词: Adult; Alleles; Animal Model; Animals; Area; Assisted Reproductive Technology; Breeding; Cell Line; Chimera organism; Code; Communities; Cryopreservation; Data; Data Coordinating Center; Databases; Deposition; ES Cell Line; Embryo; Ensure; Equipment; Excision; Fertility; Generations; Genes; Genotype; Goals; Human Biology; Informatics; International; Knock-out; Knockout Mice; Laboratories; LacZ Genes; Letters; Management Information Systems; Methods; Microinjections; Modeling; Mouse Strains; Mus; Mutant Strains Mice; Mutation; Neomycin; Partner in relationship; Pattern; Phase; Phenotype; Positioning Attribute; Process; Production; Proteins; Ramp; Reporter; Research; Resources; Schedule; Science; Services; Shipping; Ships; Specific Pathogen Frees; System; Testing; The Jackson Laboratory; Time; base; cohort; cost; data management; embryonic stem cell; experience; gene function; germ free condition; human disease; improved; innovation; innovative technologies; mammalian genome; meetings; member; mouse development; pathogen; programs; repository; reproductive; response; sperm cell; success

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.

描述(由申请者提供)：国际基因敲除小鼠联合会(IKMC)的长期目标是为哺乳动物基因组中的每个蛋白质编码基因开发一种小鼠靶向突变资源，研究社区可以利用该资源来阐明人类生物学和疾病中的基因功能。在这一框架内，KOMP^计划第一阶段的目标是在五年内产生2500条特征良好的淘汰线。该计划将在三个职能领域建立一个互动中心系统：KO菌株的生产和初步表型；基础广泛的表型；以及该计划的信息学和数据库支持。这份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，可以高效、经济地大规模执行中心的职能。该中心将利用多种JAX资源，包括JAX储存库，这是一个主要的社区老鼠资源计划；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；以及杰出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细胞培养出多达2,500个KO小鼠品系；2)测试和分析KO小鼠的存活率和繁殖力，并检查每个等位基因的LacZ表达模式；3)繁殖、冷冻保存动物和/或种质，并将其分发给KOMP^表型中心；以及4)通过利用创新技术降低向表型中心提供小鼠和种质的成本。我们将在JAX使用现有的、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)。这个项目与表型鉴定中心和DCCDB一起，承诺向科学界提供关键资源。 相关性：小鼠是理解基因功能和开发准确的人类疾病模型的关键动物模型。作为KOMP计划的一部分，这项提议旨在产生多达2500个新的突变小鼠品系，为科学界提供宝贵的资源。杰克逊实验室拥有卓越的专业知识、经验和资源，能够高效、经济地实现这一目标。