The BCM Knockout Mouse Production and Phenotyping Project (BCM KOMP2)

摘要 国际小鼠表型鉴定联盟(IMPC)的目标是开发零等位基因系资源 对于小鼠基因组中的每个蛋白质编码基因，通过标准化的、广泛的表型分析， 对每个目标基因的生物学功能进行分类。美国国立卫生研究院资助的基因敲除小鼠表型项目 (KOMP2)支持前两个阶段的无效等位基因产生和表型，有助于 一半以上的无效等位基因系是由IMPC制作和鉴定的。此应用程序描述了我们的计划 在第三阶段资助期间，贝勒医学院(BCM)将继续作为KOMP2网站。我们建议 600个蛋白质编码基因的小鼠品系的培育、冷冻保存和表型无效等位基因。我们已经开发出 一种生物信息学方法来识别和优先处理与已知或假定的人类疾病相关的基因 但对于零等位基因小鼠品系的生产，很少到没有体内功能注释。我们将生成并验证 使用已建立的CRISPR基因组编辑方法和质量控制程序的零等位基因，冷冻保存 所有零等位基因系使用公认的技术，并将种质传递到MMRRC储存库用于社区- 分布广泛。这些零等位基因系代表着科学界的黄金标准资源和 为今后的研究和翻译研究奠定了重要的基础。我们将利用我们已建立的管道来 对所有突变系进行广泛的、标准化的成体表型鉴定，并评估纯合子致死性和 胚胎表型流水线中的亚可活系。我们生成的表型数据将提供 对哺乳动物生物学以及人类疾病的遗传基础的基本新见解。所有等位基因和 表型数据将实时提交给数据协调中心，确保快速传播 将所有BCM数据提供给更广泛的生物医学科学界。我们将继续我们的生产和表型 技术开发计划，开发我们自己的新方法，并采用来自 当他们崛起的时候，社区就会出现。BCM KOMP2团队拥有成熟的专业知识、经验和资源 以高效率和高成本效益的方式实现拟议的第三阶段目标。

ABSTRACT The goal of the International Mouse Phenotyping Consortium (IMPC) is to develop a resource of null allele lines for every protein-coding gene in the mouse genome and, through standardized, broad-based phenotyping, catalog the biological function of each targeted gene. The NIH-funded Knockout Mouse Phenotyping Project (KOMP2) has supported two previous phases of null allele production and phenotyping, contributing to more than half of the null allele lines made and characterized by the IMPC. This application describes our plan for Baylor College of Medicine (BCM) to continue as a KOMP2 site during a third phase of funding. We propose to produce, cryopreserve, and phenotype null allele mouse lines for 600 protein coding genes. We have developed a bioinformatics approach to identify and prioritize genes with known or putative human disease associations but little-to-no in vivo functional annotation for null allele mouse line production. We will generate and validate null alleles using established CRISPR genome editing approaches and quality control procedures, cryopreserve all null allele lines using accepted techniques, and deliver germplasm to the MMRRC repositories for community- wide distribution. These null allele lines represent a gold standard resource for the scientific community and an important foundation for future research and translational studies. We will utilize our established pipelines to perform broad-based, standardized adult phenotyping on all mutant lines and assess homozygous lethal and subviable lines in an embryonic phenotyping pipeline. The phenotyping data we generate will provide fundamental new insights into mammalian biology as well as the genetic bases of human disease. All allele and phenotype data will be submitted in real time to the Data Coordination Center, ensuring the rapid dissemination of all BCM data to the wider biomedical scientific community. We will continue our production and phenotyping technology development programs, developing our own new approaches and adopting new methodologies from the community as they arise. The BCM KOMP2 team has the established expertise, experience, and resources to meet the proposed phase 3 goals in an efficient and cost-effective manner.