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.

抽象的 小鼠和人类共享大约 20,000 个基因。迄今为止，其中四分之一以上的数据很少 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 小鼠和人类基因集，小鼠产生的遗传数据通常可以推断出人类基因功能。 小鼠和人类之间具有共同功能的基因的小鼠模型可以产生新的模型 疾病，这对于药物筛选、临床前研究以及更深入地了解生物学和 发病机制。基因敲除小鼠表型分析程序 (KOMP2) 的目标是生成 携带全基因组小鼠基因敲除 (KO) 的小鼠，并对小鼠进行广泛的 基于表型分析。 JAX KOMP2第3期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 600 个小鼠系并对其进行表型分析 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，并将重点关注以下方面：有人类直系同源物，尚未 以前被淘汰，没有注释或注释很差，有很大的社区需求并与其他集成 NIH 支持计划或预计将在选定的途径中发挥作用。为了保证随时访问 社区中，当小鼠在架子上还活着时，我们会将它们运送给外部调查人员，并将线路存放在 小鼠突变体区域资源中心 (MMRRC) 存储库以供将来使用。 将对所有 600 个品系的 17 周龄以下年轻成年小鼠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。我们将评估 体重和成分、行为、心血管、代谢、眼部和生理参数。 根据 KOMP2 当前阶段生成的数据，我们预计大约 30% 的品系无法生存。我们 将在三个胚胎时间点使用高通量成像方式表征非存活突变体。 根据之前的数据，我们还预计大约 7% 的品系是不育的。直接生育力测试 评估每个性别的生育力将对所有未能从纯合子产生后代的品系进行 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中心 支持 KOMP2 和 IMPC 的协调中心 (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。