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)

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要小鼠和人类共享大约 20,000 个基因。迄今为止，其中一半以上的数据很少基因，近三分之一没有功能注释。由于两者之间的相似度很高小鼠和人类基因组，小鼠产生的遗传数据通常可以推断到人类基因功能。具有小鼠和人类之间共同功能的基因的小鼠模型可以产生新的模型人类疾病，这有助于药物筛选、临床前研究和更深入地了解生物学及发病机制。基因敲除小鼠表型分析程序 (KOMP2) 的目标是生成品系对全基因组小鼠基因进行敲除 (KO) 的小鼠，并对小鼠进行基础广泛的表型分析。杰克逊实验室 (JAX) 提议合并其目前资助的 KOMP2 小鼠生产和冷冻保存 (U42) 以及基因敲除小鼠表型分析 (U54) 操作作为 RFA-RM-15-017 有限竞赛：淘汰小鼠生产和表型项目 (UM1)。 JAX KOMP2建议使用尖端且经济高效的Cas9 RNA引导核酸酶（Cas9 RGN，也称为 CRISPR/Cas9) 技术，用于生成、繁殖、冷冻保存 1500 个小鼠系并进行表型分析项目期间。将继续努力改进Cas9 RGN技术以降低成本，提高靶向效率，并创建更复杂的突变等位基因。基因将被协调选择与我们的 KOMP2 和 IMPC 合作伙伴合作，并将重点关注那些注释较差、具有显着意义的基因社区需求并与其他 NIH 支持计划相结合，以及预测在选择中发挥作用的基因途径。为了保证能够随时进入社区，我们会将小鼠运送给外部调查人员，同时他们架子上还活着，并将这些品系存入小鼠突变体区域资源中心 (MMRRC) 存储库以供将来使用。将对 18 周龄以下的幼年动物的所有 1500 个品系进行广泛的表型分析使用国际小鼠表型联盟 (IMPC) 要求的和 JAX 特定的协议对小鼠进行分析。我们将评估体重和成分、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，我们将使用相同的流程对 20%（总共 300 个）的表型进行检测 15-18个月龄之间的界限。根据 KOMP2 当前阶段生成的数据，我们预计大约 30% 的生产线将无法生存。我们将使用高通量表征无活力的突变体三个胚胎时间点的成像方式。所有数据均来自胚胎、幼年和成年小鼠将被快速存入支持 KOMP2 和 IMPC 的数据协调中心 (DCC)。最后，JAX 将与 KOMP2 区域网络以及 KOMP2 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、会议演示、网络活动和社交媒体向国际社会展示。