KOMP2-Phase 2 Production and Phenotyping by the DTCC Consortium

DTCC财团的KOMP2-Phase2生产和表型鉴定 摘要 这项申请旨在竞争性地为DTCC财团第二阶段参与 基因敲除小鼠生产和表型(KOMP2)项目。DTCC联盟成员中心 包括加州大学戴维斯分校(UCD；牵头机构)、#年的表型基因组学中心 多伦多、奥克兰儿童医院研究所(CHORI)和查尔斯河实验室 国际公司(CRL)。我们建议利用我们的经验，继续使用经过良好测试的协调 KOMP2-Phase1成功实施的管理、战略、协议和流程，以增加， 扩大和加强KOMP2-Phase2的活动。在KOMP2-阶段1中，DTCC已完成(或正在按计划进行 在阶段结束前完成1)所有既定目标并实现所有指标(例如，加速微量注射 ES细胞克隆，提高生殖系突变体的产率，提前完成生产时间表， 完成表型鉴定的跟踪)超出了我们工作范围所需的目标和目标 DTCC在KOMP2-Phase2中提出。在这次更新中，DTCC建议对男性和女性进行表型 至少1,500个突变纯合子(Hom)或杂合(Het)小鼠品系的队列 Cas9 RNA引导核酸酶(Cas9 RGN)和相应的年龄和性别匹配的野生型(WT)对照 3条表型管道(胚胎、青少年和成人)，使用所有IMPC必需的和几个可选的印记- 建立了跨多个器官系统的测试。在应用基于中心的工艺质量保证之后， 和产品(鼠标和数据)的质量控制，所有数据(包括元数据)将立即上传到 数据协调中心(DCC)，然后是鼠标表型信息基础设施(MPI2) 通过IMPC门户网站(www.impc.org)向公众开放。为了提高生成老鼠的效率 线路和数据，DTCC成员中心将进行技术开发项目，以改进和增加 生产效率(例如，高效RGN介导的目标定位、LacZ敲击插入)和试点 表型测试(例如，自动家庭笼子监测和微核[基因组不稳定性])以确定 它们在我们的青少年和成人渠道中的附加值、再现性和成本效益。表型分型 技术开发活动将包括一种新的基于定量试剂盒的蛋白质组分析和代谢组学 突变小鼠品系的概况，承诺增加显著的科学价值和提高研究效用 表型数据。此外，DTCC将通过征集提名的方式积极参与科学界的工作 对于要研究的基因(例如，成体管道)，在生产和生产KOMP2小鼠的同时，对请求的响应进行优先排序 表型鉴定正在进行中，并在沉积到突变小鼠资源和研究中心后 (www.mmrrc.org)，并提供增值服务和协作(例如，PAR-13-231表型 胚胎死亡，组织病理学支持贝勒-哈威尔KOMP2-阶段2联盟)。除了输入之外 从NIH项目官员那里，DTCC将以DTCC小组的意见和建议为指导 专家顾问、NIH KOMP2科学顾问委员会和IMPC科学顾问小组。这个 DTCC将参加KOMP2和IMPC联盟的所有活动，包括每月举行的电话会议， 小组委员会工作组和会议、国家和国际会议、实地考察和其他 相关活动。为了最大限度地提高科学产出和对生物医学研究的影响，DTCC还 承诺通过以下方式协调其努力与这些生产、表型和研发工作 其他由KOMP2资助的参与者。总而言之，到5年结束时，DTCC将做出贡献 至少1500个突变小鼠品系的经基因确认的小鼠和经过验证的基于广泛的表型数据 按时并在预算范围内支持KOMP2计划和IMPC工作，以便在功能上说明 哺乳动物的基因组。

KOMP2-Phase2 Production and Phenotyping by the DTCC Consortium ABSTRACT This application is to competitively renew funding for the DTCC Consortium’s Phase2 participation in the Knockout Mouse Production and Phenotyping (KOMP2) Project. Member centers of the DTCC Consortium include the University of California Davis (UCD; lead institution), The Centre for Phenogenomics (TCP) in Toronto, the Children’s Hospital Oakland Research Institute (CHORI), and Charles River Laboratories International Inc (CRL). We propose to leverage our experience and continue to use well-tested coordinated management, strategies, protocols, and processes successfully implemented for KOMP2-Phase1 to increase, expand, and enhance activities for KOMP2-Phase2. In KOMP2-Phase1, the DTCC completed (or is on track to complete by the end of Phase1) all established goals and achieve all metrics (e.g., accelerated microinjection of ES cell clones, increased production rate of germline mutants, early completion of production timeline, on- track for completion of phenotyping) that exceed objectives and goals required for the scope of work we propose by the DTCC in KOMP2-Phase2. In this renewal, the DTCC proposes to phenotype male and female cohorts for at least 1,500 mutant homozygous (HOM) or heterozygous (HET) mouse lines produced using Cas9 RNA-guided nuclease (Cas9 RGN) and corresponding age and sex –matched wildtype (WT) controls in 3 phenotyping pipelines (Embryo, Juvenile, and Adult) using all IMPC-required and several optional IMPReSS- established tests across multiple organ systems. After applying center-based quality assurance of processes, and quality control of products (mice and data), all data (including meta-data) will be uploaded without delay to the Data Coordination Center (DCC) and then the Mouse Phenotyping Informatics Infrastructure (MPI2) for public access through the IMPC web portal (www.impc.org). To enhance the efficiency of generating mouse lines and data, DTCC member Centers will conduct technology development projects to improve and increase efficiency of production (e.g. high-efficiency RGN-mediated targeting, lacZ knockin insertion) and pilot phenotyping tests (e.g. automated home cage monitoring and micronucleus [genome instability]) to establish their added-value, reproducibility, and cost-effectiveness in our Juvenile and Adult Pipelines. Phenotyping technology development activities will include a novel quantitative kit-based proteomic assay, and metabolomic profiling of mutant mouse lines that promise to add significant scientific value and increase research utility of phenotyping data. In addition, the DTCC will actively engage the scientific community by seeking nominations for genes to study (e.g., Adult Pipeline), prioritize responses to requests for KOMP2 mice while production and phenotyping is in progress and after deposition into the Mutant Mouse Resource and Research Center (www.mmrrc.org), and provide value-added services and collaborations (e.g., PAR-13-231 Phenotyping Embryo Lethals, histopathology support to the Baylor-Harwell KOMP2-Phase2 Consortium). In addition to input from NIH Program Officers, the DTCC will be guided by advice and recommendations of the DTCC Panel of Expert Advisors, the NIH KOMP2 Scientific Advisory Board, and the IMPC Panel of Scientific Consultants. The DTCC will participate in all KOMP2 and IMPC consortium activities, including monthly teleconferences, subcommittee working groups and meetings, national and international conferences, site visits, and other relevant activities. In order to maximize the scientific output and effect on biomedical research, the DTCC also commits to coordinate its efforts with those production, phenotyping, and research and development efforts by other KOMP2-funded participants. In summary, by the end of 5 years, the DTCC will have contributed genotype-confirmed mice and validated broad-based phenotyping data for at least 1,500 mutant mouse lines on time and within budget in support of the KOMP2 program and the IMPC effort to functionally annotate the mammalian genome.