KOMP PHASE II MOUSE PRODUCTION AND CRYOPRESERVATION: AIDS

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 简介(申请人提供)：与爱滋病有关。加州大学戴维斯分校、多伦多表型基因组学中心(TCP)和Charles River实验室共同提出的这项提案被称为“DTC”财团，旨在建立和运营一个基因敲除的老鼠计划第二阶段(K0MP2)生产中心。我们生产中心的目标是每年从IKMC ES细胞中产生500个缺失突变小鼠系，对缺失和条件就绪突变的种质进行初步分析和冷冻保存，并通过IKMC存储库向表型中心和研究社区提供小鼠和数据。我们的长期目标是制定和实施提高生产能力的生产策略，以便在确保产品质量和可靠性的同时，以较低的成本每年生产多达1,250个突变小鼠品系。总而言之，我们在处理和将ES细胞转化为小鼠、质量控制、测试和分析、种质超低温保存和恢复以及育种群体的快速扩张方面的记录，特别是在C57BL/6N背景下，完全支持我们每年生产全部500个突变小鼠品系的计划。我们参与K0MP1将使我们能够立即开始向表型鉴定中心提供高质量、可靠和一致的产品。简而言之，UCD将启动生产管道，并扩展和验证所有突变的IKMC ES细胞。完全合格的ES细胞将被转化为嵌合体，用于育种到种系确认的小鼠品系，必要时切除NEO盒式磁带和/或关键外显子，LacZ分析，以及条件等位基因种质的冷冻保存。UCD和TCP随后将把表达Cre切除后缺失等位基因的杂合子小鼠运送到CRL进行大规模繁殖和扩张，纯合子存活率、生育力和繁殖力测试，以及胚胎冷冻保存。根据要求，CRL将准备向K0MP2表型鉴定中心运送冷冻胚胎、繁殖对或完整的队列。UCD、TCP和CRL还应将小鼠和种质保存回UCD的KOMP储存库，以便存档和分发。此外，UCD和TCP将开展几个与资源相关的开发项目，以加强对突变小鼠品系的处理、生产和分析。 相关性：转基因小鼠是了解人类疾病的优秀研究模型。这项研究将为我们理解基因在疾病过程中的作用增加重要的知识。有了这些知识，我们可以更好地确定病因、治疗和设计针对疾病的预防战略，从而使国家人口更加健康。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. DESCRIPTION (provided by applicant): AIDS related. This proposal from UC Davis, Toronto Centre for Phenogenomics (TCP), and Charles River Laboratories, together known as the "DTC" consortium, seeks to establish and operate a Knock-Out Mouse Program Phase 2 (K0MP2) Production Center. The purpose of our Production Center will be to generate 500 deletion mutant mouse lines from IKMC ES cells each year, conduct preliminary analyses on and cryopreserve germplasm from both deletion and conditional-ready mutants, and make mice and data readily available to phenotyping centers and to the research community through IKMC repositories. Our long-term objective is to develop and implement production strategies that increase throughput to facilitate production of up to ~1,250 mutant mouse lines annually at lower cost while ensuring product quality and reliability. Together, our track record of handling and converting ES cells to mice, quality control, testing and analyses, germplasm cryopreservation and recovery, and rapid expansion of breeding colonies, especially on the C57BL/6N background, fully supports our plan to produce all 500 mutant mouse lines per year. Our participation in K0MP1 will enable us to immediately begin to deliver high quality, reliable, and consistent product to the phenotyping centers. Briefly, UCD shall initiate the production pipeline and expand and verify all mutant IKMC ES cells. Fully qc'd ES cells will be converted into chimeras for breeding to germline confirmed mouse lines, excision of the neo cassette and/or critical exon as necessary, lacZ analysis, and cryopreservation of germplasm from conditional alleles. UCD and TCP will then ship heterozygous mice expressing the post- CRE excision deletion allele to CRL for large-scale breeding and expansion, homozygous viability, fertility, and fecundity testing, and embryo cryopreservation. Upon request, CRL will be prepared to ship frozen embryos, breeding pairs, or full cohorts to K0MP2 Phenotyping Centers. UCD, TCP, and CRL shall also deposit mice and germplasm back to the KOMP Repository at UCD for archiving and distribution. In addition, UCD and TCP will conduct several resource-related development projects to enhance handling, production, and analysis of mutant mouse lines. RELEVANCE: Genetically altered mice serve as excellent research models to understand human disease. This study will add significant knowledge to our understanding of the roles of genes in disease processes. With this knowledge, we can better identify causes, treat, and design prevention strategies against diseases, leading to a healthier national population.