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: 10517971
• 负责人: ROBERT E BRAUN
• 金额: $ 272.57万
• 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 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10517971
• 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; International; Internet; Knock-out; Knockout Mice; Knowledge; Laboratories; Lead; 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; Ships; Standardization; Technology; Testing; The Jackson Laboratory; Time; United States National Institutes of Health; Work; aged; animal data; animal resource; base; cohort; cost; cost effective; endonuclease; experience; 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; social media; tool; young adult

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.

摘要 老鼠和人类共有大约20，000个基因。迄今为止，其中四分之一以上的数据很少 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。 小鼠和人类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，这是有用的药物筛选，临床前研究，并更深入地了解生物和 发病机制敲除小鼠表型分析程序（KOMP 2）的目标是产生小鼠表型分析的细胞系。 携带小鼠基因的全基因组集合的敲除（科斯）的小鼠，并使小鼠经受广泛的 基于表型分析。 JAX KOMP 2第3阶段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CRISPR/Cas9）技术，以产生、繁殖、冷冻保存和表型600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，并将重点放在那些有人类直系同源物， 以前被淘汰，没有注释或注释不佳，有重要的社区需求，并与其他社区整合 NIH支持计划，或预测在选择途径中发挥作用。为了保证随时可以进入 社区，我们将运送小鼠到外部调查人员，而他们是活的架子上，并存款线到 小鼠突变区域资源中心（MMRRC）储存库供未来使用。 将对所有600个品系的小鼠进行17周龄以下的年轻成年小鼠的广泛表型分型 使用国际小鼠表型鉴定协会（IMPC）要求的和JAX特定的协议。我们将评估 体重和组成，以及行为、心血管、代谢、眼部和生理参数。 根据KOMP 2当前阶段生成的数据，我们预计约有30%的品系是不可存活的。我们 将在三个胚胎时间点使用高通量成像模式来表征非存活突变体。 根据以前的数据，我们还预计大约7%的品系是不育的。直接进行生育力测试， 评估每种性别的生育力将在未能从纯合产生后代的所有品系上进行， 纯合交配胚胎和成年小鼠产生的所有数据将快速存入数据库。 协调中心（DCC）支持KOMP 2和IMPC。 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，并广泛和公开地传播我们的研究结果， 通过出版物、在会议上的发言、网络活动和社交媒体向国际社会提供信息。