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：
10683282
负责人：
ROBERT E BRAUN
金额：
$ 233.95万
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 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10683282
关键词：
Adult Age Alleles Archives Award Behavior Biological Biological Models Biology Body Composition Body Weight Breeding CRISPR/Cas technology Cardiovascular system Catalogs Collection Communities Complement Complex Coupled Cryopreservation Data Data Coordinating Center Databases Deposition Development Disease Disease model Drug Screening Embryo Encyclopedias Engineering Ensure Faculty Fertility Future Generations Genes Genetic Genotype Goals Guide RNA Health Human Infertility Infrastructure Institution International Internet Knock-out Knockout Mice Knowledge Laboratories Leadership Link Metabolic Methods Mission Mus Mutant Strains Mice Orthologous Gene Partner in relationship Pathway interactions Phase Phenotype Physiological Production Protocols documentation Public Health Publications Recording of previous events Reproducibility Research Research Personnel Resources Role Services Standardization Technology Testing The Jackson Laboratory Time United States National Institutes of Health Work aged animal data animal resource cohort cost cost effective endonuclease experience fortification gene function genome-wide human disease human model imaging modality improved innovation meetings member mouse genetics mouse genome mouse model multidisciplinary mutant new technology novel novel strategies nuclease offspring preclinical study programs repository sex skills social media tool transmission process young adult

项目摘要

ABSTRACT Mice and humans share approximately 20,000 genes. To date, little data exists for more than a quarter 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 humans can lead to new models of 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. JAX KOMP2 phase 3 proposes to use cutting-edge and cost-effective Cas9 RNA-guided nuclease (Cas9 RGN, also called CRISPR/Cas9) technology to generate, breed, cryopreserve and phenotype 600 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 that; have human orthologs, have not been previously knocked out, have no or poor annotation, have significant community demand and integrate with other NIH-support programs, or are 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 young adult mice up to 17 weeks of age will be performed on all 600 lines of mice using International Mouse Phenotyping Consortium (IMPC)-required and JAX-specific protocols. We will assess body weight and composition, and behavior, cardiovascular, metabolic, ocular and physiological parameters. 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. Based on previous data we also expect approximately 7% of the lines to be infertile. Direct fertility testing to assess the fertility of each sex will be performed on all lines that fail to generate offspring from homozygous by homozygous matings. All data generated from embryonic 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.

抽象的小鼠和人类共享约20,000个基因。迄今为止，数量很少的数据超过四分之一基因和近三分之一没有功能注释。因为小鼠和人类基因集，在小鼠中产生的遗传数据通常可以推断到人基因功能。小鼠和人之间具有共同功能的基因的小鼠模型可以导致新的模型疾病，可用于药物筛查，临床前研究以及对生物学和生物学的更深入了解疾病机制。淘汰鼠标表型程序（KOMP2）的目标是生成线条携带淘汰的小鼠（KOS），用于全基因组的小鼠基因收集，并使小鼠宽阔基于表型。 JAX KOMP2第3阶段建议使用尖端和成本效益的Cas9 RNA引导的核酸酶（CAS9 RGN，也称为CRISPR/CAS9）在期间生成，繁殖，冷冻水果和表型600行的技术项目期。将继续努力改善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。基因将在协调中选择与我们的KOMP2和IMPC合作伙伴一起，并将专注于这些合作伙伴；有人类直系同源物以前被淘汰，没有或没有差注释，有巨大的社区需求并与其他 NIH支持程序，或预计将在某些途径中起作用。保证准备就可以访问社区，我们将把老鼠运送到外部调查人员的架子上，并将线路存放到小鼠突变区域资源中心（MMRRC）存储库，以供将来使用。在所有600行小鼠上，将对长达17周龄的年轻小鼠对年轻小鼠进行广泛的表型使用国际小鼠表型联盟（IMPC）率特定的方案。我们将评估体重和组成以及行为，心血管，代谢，眼和生理参数。基于从KOMP2的当前阶段生成的数据，我们预计约有30％的线条是不可行的。我们将使用高通量成像方式在三个胚胎时间点使用高通量成像方式来表征不可生存的突变体。基于先前的数据，我们还期望大约7％的线不育。直接生育测试评估每种性别的生育能力将在所有无法从纯合子中产生后代产生后代的行执行纯合子垫。由胚胎和成年小鼠产生的所有数据都将迅速沉积到数据中支持KOMP2和IMPC的协调中心（DCC）。最后，JAX将与Komp2区域网络合作，并与 IMPC共享协议，创新和新技术，并广泛，公开地传播我们的发现国际社会通过出版，会议，网络活动和社交媒体的演讲。