KOMP Phase II Mouse Phenotyping

描述(申请人提供)：这个K0MP2项目的目标是在五年内每年提供200个KOMP突变小鼠品系的综合表型。这将向研究界提供关于基因功能的重要新信息，特别是对于未注释的基因，并将加速我们对疾病的遗传基础的理解。这一目标将由一个在突变小鼠生产、表型鉴定、技术创新和合作项目方面拥有显著记录的财团来实现。UCD-TCP-CHORI联盟由加州大学戴维斯分校(UCD)的老鼠生物学项目(UCD)、多伦多表型基因组学中心(TCP)和奥克兰儿童医院(CHORI)组成。在这项应用中，我们建议每年对200个纯合子K0MP2突变株进行表型鉴定：150个成年突变株将使用单一管道方法在UCD-CHORI位点进行表型鉴定，并将分子表型整合到评估中；50个亚存活株系将在TCP进行表型鉴定，同时对胚胎致死系的子集进行杂合子的成年表型鉴定。此外，TCP将使用成熟的挑战测试来评估纯合子突变的成人肺功能，该测试适用于成年可存活突变系的子集。将协调UCD-TCP-CHORI联盟成员共享的成人表型的方法、程序和最佳做法。该计划与国际老鼠表型鉴定联盟的建议很好地结合在一起，但我们建议的变化将节省大量资源，并提供丰富的新颖数据集。创新包括：A)用于成体可存活系的单一管道方法；B)结合多种成像方式的密集和详细的胚胎致死表型；C)定向亚可活突变表型；以及D)纯合子成人的智能转录组分析。此外，我们还提出了一项密集的技术开发计划，以应用DNA测序技术中的革命来提炼、定制和优化用于转录组分析的RNAseq，并探索其他分子表型，如microRNA图谱和代谢组学，以便将来纳入表型筛选。

DESCRIPTION (provided by applicant): The objective of this K0MP2 project is to provide comprehensive phenotyping of 200 KOMP mutant mouse lines each year for five years. This will provide significant new information regarding gene function, especially for un-annotated genes, to the research community and will accelerate our understanding of the genetic basis of disease. This goal will be achieved by a consortium having a significant track-record in mutant mouse production, phenotyping, technical innovation and working together on collaborative projects. The UCD-TCP-CHORI consortium consists of the Mouse Biology Program at U. California Davis (UCD), the Toronto Center for Phenogenomics (TCP), and Children's Hospital of Oakland Research Institution (CHORI). In this application, we propose phenotyping 200 homozygous K0MP2 mutant lines each year: 150 adult mutant lines will be phenotyped at the UCD - CHORI site using a single pipeline approach and integrating molecular phenotyping into the assessment; and 50 sub-viable lines will be phenotyped at TCP along with adult phenotyping of heterozygotes for a subset of the embryonic lethal lines. In addition, TCP will assess homozygous mutant adult lung function using a well established challenge test in a subset of the adult-viable mutant lines. Methods, procedures-and best practices for adult phenotypes shared among the UCD-TCP-CHORI consortium members will be harmonized. The plan integrates well with the recommendations of the International Mouse Phenotyping Consortium, but we have recommended changes that will save significant resources, and provide rich novel datasets. Innovations include: A) a single pipeline approach for the adult-viable lines; B) intensive and detailed embryonic lethal phenotyping that incorporates several imaging modalities; C); targeted sub-viable mutant phenotyping; and D) intelligent transcriptome analyses in homozygous adult. Also, we propose an intensive technology development program to apply the revolutions in DNA sequencing technology to refine, tailor and optimize RNAseq for transcriptome analyses and to explore other molecular phenotypes such as micro RNA profiling, and metabolomics for future incorporation into the phenotype screens.