The Jackson Laboratory Knockout Mouse Production and Phenotyping Project (JAX KOMP2)

项目摘要 杰克逊实验室（JAX）敲除小鼠表型分型项目第2阶段（KOMP 2）的目标是 有助于建立一个全面的哺乳动物基因功能目录的国际努力， 通过创建基因敲除和执行高通量表型分析管道。在JAX， Early Adult Pipeline（EAP）用于对17周龄以下的小鼠进行表型分析，目标是筛选1000只小鼠。 在目前的5年项目期结束前，此外，在此授予期间，150个子集 品系正在老化至18个月，并使用晚期成体管道（Late Adult Pipeline，简称LIPV）进行表型分析，以检测与年龄相关的 表型无活力和亚活力品系也正在使用专门的胚胎表型分析进行表型分析 管道2020年3月11日，世界卫生组织（WHO）宣布， 呼吸道综合征冠状病毒2型（SARS-CoV-2）疾病（COVID-19）是一种全球性大流行病。因此 这一大流行病的后果是，我们减少了实现我们最初目标所需的现有人员和服务。 大流行期间的限制减少了JAX KOMP 2中心的繁殖和表型分析活动。 我们保持了较低水平的群体繁殖，并减少了小鼠的表型。另外为了 为了最大限度地保持物理距离，并以减轻病毒传播和扩散的方式运作，我们 需要工作人员之间密切接触的程序已停止。由于大流行，我们的总数不足 预计已进行表型分型的品系数量和所有试验的全队列品系数量。的 请求在当前项目期之后延长供资期限的目的是完成最初目标 1000个基因敲除小鼠品系的实验。我们预计，在最初的赠款期结束时， 860个品系的小鼠将完成EAP管道，并且来自所有150个品系的表型分析数据将被 收集。这一资助扩展的具体目标是从剩余的140个中繁殖和繁殖EAP 鼠标线使我们的总数达到1000行。大约70%的生产线（约100条生产线）将完全可行， 因此将被表型为纯合子。其余30%的细胞系（约40个细胞系）为非存活细胞系或 亚存活的，因此将作为杂合子进入EAP。这40行也将用于填充我们的 胚胎致死表型分析管道。将对EAP数据和EAP数据进行QC分析并导出 国际小鼠表型鉴定联盟数据协调中心

PROJECT SUMMARY The goal of The Jackson Laboratory (JAX) Knockout Mouse Phenotyping Project phase 2 (KOMP2) is to contribute to the international effort of creating a comprehensive catalog of mammalian gene function obtained through the creation of gene knockouts and execution of a high-throughput phenotyping pipeline. At JAX an Early Adult Pipeline (EAP) is being used to phenotype mice up to 17 weeks of age with a goal of screening 1,000 mutant lines by the end of the current 5-year project period. Further, during this award period, a subset of 150 lines is being aged out to 18 months and phenotyped using a Late Adult Pipeline (LAP) to detect age-associated phenotypes. Nonviable and subviable lines are also being phenotyped using specialty embryonic phenotyping pipelines. On March 11, 2020 the World Health Organization (WHO) declared the outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) disease (COVID-19) a world-wide pandemic. A consequence of the pandemic has been a reduction of our available personnel and services needed to reach our original goals. Restrictions during the pandemic have curtailed the breeding and phenotyping activity on the JAX KOMP2 center. We have maintained a lower level of breeding of cohorts and reduced phenotyping of mice. In addition, in order to maximize physical distancing and operate in a way that mitigates viral transmission and spread, we have discontinued procedures that require close contact between staff. Due to the pandemic, we are short of the total number of lines projected to have been phenotyped and on the number of lines with full cohorts for all tests. The purpose of this request for a funded extension beyond the current project period is to complete the initial goals of creating and phenotyping 1000 lines of KO mice. We project that at the end of the original grant period that 860 lines of mice will have completed the EAP pipeline and that LAP phenotyping data from all 150 lines will be collected. The Specific Aims of this funded extension are to breed and populate the EAP from the remaining 140 mouse lines to bring our total to 1000 lines. Approximately 70% of the lines (~100 lines) will be fully viable and will therefore be phenotyped as homozygotes. The remaining 30% of the lines (~40 lines) be non-viable or subviable and will therefore enter the EAP as heterozygotes. These 40 lines will also be used to populate our embryo lethal phenotyping pipeline. QC analysis will be performed on both the EAP and LAP data and exported to the Data Coordination Center of the International Mouse Phenotyping Consortium.