The Jackson Laboratory Knockout Mouse Production and Phenotyping Project (JAX KOMP2)

杰克逊实验室基因敲除小鼠生产和表型项目 (JAX KOMP2)

• 批准号: 9754888
• 负责人: ROBERT E BRAUN
• 金额: $ 624.12万
• 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: 2011-09-16 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9754888
• 关键词: Adolescent; Adult; Age; Age-Months; Alleles; Archives; Award; Behavioral; Biological; Biological Models; Biology; Body Composition; Body Weight; Breeding; CRISPR/Cas technology; Catalogs; Collection; Communities; Complement; Complex; Coupled; Cryopreservation; Data; Data Coordinating Center; Databases; Deposition; Development; Disease; Drug Screening; Embryo; Encyclopedias; Engineering; Ensure; Faculty; Fertility; Funding; Future; Generations; Genes; Genetic; Genotype; Goals; Guide RNA; Health; Human; Infrastructure; International; Internet; Knock-out; Knockout Mice; Knowledge; Laboratories; Lead; Leadership; Link; Metabolic; Mission; Mus; Mutant Strains Mice; Pathway interactions; Phase; Phenotype; Physiological; Production; Protocols documentation; Public Health; Publications; Recording of previous events; Reproducibility; Research; Research Personnel; Resources; Role; Services; Ships; Standardization; Technology; Testing; The Jackson Laboratory; Time; United States National Institutes of Health; Work; age related; aged; animal data; animal resource; base; cognitive function; cohort; cost; cost effective; endonuclease; experience; gene function; genome-wide; human disease; human model; imaging modality; improved; innovation; juvenile animal; meetings; member; men; mouse genome; mouse model; multidisciplinary; mutant; new technology; novel; novel strategies; nuclease; operation; preclinical study; programs; repository; social media; tool

PROJECT SUMMARY/ABSTRACT Mice and humans share approximately 20,000 genes. To date, little data exists for more than half of these genes and nearly one third have no functional annotation. Because of the high degree of similarity between the mouse and human gene set, genetic data generated in mice can often be extrapolated to human gene function. Mouse models of genes with common functionality between mice and men can lead to new models of human disease, which are useful for drug screening, preclinical studies and deeper understanding of biological and disease mechanism. The goal of the Knockout Mouse Phenotyping Program (KOMP2) is to generate lines of mice that carry knockouts (KOs) for a genome-wide collection of mouse genes and subject the mice to broad based phenotyping. The Jackson Laboratory (JAX) proposes to merge its currently funded KOMP2 Mouse Production and Cryopreservation (U42) and Knockout Mouse Phenotyping (U54) operations as part of RFA-RM-15-017 Limited Competition: Knockout Mouse Production and Phenotyping Project (UM1). JAX KOMP2 proposes to use cutting-edge and cost-effective Cas9 RNA-guided nuclease (Cas9 RGN, also called CRISPR/Cas9) technology to generate, breed, cryopreserve and phenotype1500 lines of mice during the project period. Continued effort will be made to improve the Cas9 RGN technology so as to reduce costs, increase targeting efficiency, and create more complex mutant alleles. Genes will be selected in coordination with our KOMP2 and IMPC partners, and will focus on those with poor annotation, genes that have significant community demand and integrate with other NIH-support programs, and genes predicted to function in select pathways. To guarantee ready access to the community, we will ship mice to outside investigators while they are alive on the shelf and deposit the lines into the Mouse Mutant Regional Resource Center (MMRRC) repositories for future use. Broad based phenotyping on juvenile animals up to 18 weeks of age will be performed on all 1500 lines of mice using International Mouse Phenotyping Consortium (IMPC)-required and JAX-specific protocols. We will assess body weight and composition, metabolic and physiological parameters, and behavioral and cognitive function. To detect age-dependent phenotypes, we will use the same pipeline to phenotype 20% (300 total) of the lines between 15-18 months of age. Based on data generated from the current phase of KOMP2, we expect about 30% of lines to be non-viable. We will characterize the non-viable mutants using high-throughput imaging modalities at three embryonic time points. All data generated from embryonic, juvenile and adult mice will be rapidly deposited into the Data Coordination Center (DCC) that supports KOMP2 and the IMPC. Lastly, JAX will work collaboratively with the KOMP2 Regional Network and with member organizations of the IMPC to share protocols, innovation, and new technology and to broadly and openly disseminate our findings to the international community through publication, presentations at meetings, web activities and social media.

项目总结/摘要 老鼠和人类共有大约20，000个基因。迄今为止，其中一半以上的数据很少 近三分之一的基因没有功能注释。由于两者之间的高度相似性， 小鼠和人类基因集，在小鼠中产生的遗传数据通常可以外推到人类基因 功能小鼠模型的基因与小鼠和男性之间的共同功能可以导致新的模型， 人类疾病，这是有用的药物筛选，临床前研究和更深入地了解生物 疾病机制。敲除小鼠表型分析程序（KOMP 2）的目标是产生品系 对携带全基因组小鼠基因集合的敲除（科斯）的小鼠， 广泛的表型分析。杰克逊实验室（JAX）建议将其目前资助的KOMP 2 小鼠生产和冷冻保存（U42）和敲除小鼠表型分析（U 54）操作，作为 RFA-RM-15-017有限竞争：敲除小鼠生产和表型分析项目（UM 1）。 JAX KOMP 2提出使用尖端且具有成本效益的Cas9 RNA引导的核酸酶（Cas9 RGN，也称为Cas9 RNA引导的核酸酶）。 称为CRISPR/Cas9）技术，以产生，繁殖，冷冻保存和表型1500系小鼠， 项目期间。将继续努力改进Cas9 RGN技术以降低成本， 增加靶向效率，并产生更复杂的突变等位基因。基因将被选择在协调 与我们的KOMP 2和IMPC合作伙伴，并将重点放在那些注释不佳的基因上， 社区需求和与其他NIH支持计划的整合，以及预测在选择中发挥作用的基因 途径。为了保证随时进入社区，我们将把老鼠运送给外部调查人员， 存活在货架上，并将品系存款到小鼠突变区域资源中心（MMRRC） 仓库供将来使用。 将对所有1500个品系的18周龄以下幼龄动物进行广泛的表型分析。 使用国际小鼠表型鉴定协会（IMPC）要求的和JAX特异性的方案对小鼠进行检测。我们将 评估体重和组成、代谢和生理参数以及行为和认知 功能为了检测年龄依赖性表型，我们将使用相同的管道对20%（共300个）的 年龄在15-18个月之间。根据KOMP 2当前阶段生成的数据，我们 预计约30%的线路将无法生存。我们将使用高通量技术来表征非存活突变体， 在三个胚胎时间点的成像模式。所有数据均来自胚胎、幼年和成年小鼠 将被迅速存入支持KOMP 2和IMPC的数据协调中心（DCC）。 最后，JAX将与KOMP 2区域网络和国际原子能机构的成员组织合作， IMPC分享协议、创新和新技术，并广泛公开地传播我们的发现 通过出版物、会议发言、网络活动和社交媒体向国际社会宣传。