Genetically Modified Mouse Core

转基因小鼠核心

• 批准号: 10200773
• 负责人: DOUGLAS J EPSTEIN
• 金额: $ 16.91万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10200773
• 关键词: Address; American; Animals; Bacterial Artificial Chromosomes; Biological Models; Biopsy; CRISPR/Cas technology; Collaborations; Color; Complement; Cost efficiency; Cryopreservation; DNA; Data; Derivation procedure; Development; Digestive System Disorders; Disease; ES Cell Line; Embryo; Embryo Transfer; Equipment; Female; Fertility; Fertilization; Fertilization in Vitro; Fostering; Freezing; Gastrointestinal tract structure; Gene-Modified; Generations; Genetic; Genotype; Goals; Housing; Implant; In Vitro; Incidence; Individual; Infection; Injections; Institution; International; Investigation; Knock-in; Laboratories; Lasers; Liver; Liver diseases; Messenger RNA; Methods; Microinjections; Modeling; Modification; Molecular; Monitor; Mothers; Mouse Strains; Mus; Mutation; Operative Surgical Procedures; Pancreas; Pancreatic Diseases; Pathogenicity; Pathologic; Pathway interactions; Pennsylvania; Physiological; Population; Productivity; Protocols documentation; Quality Control; Reproducibility; Research; Research Personnel; Resource Sharing; Resources; Secure; Services; Shipping; Site; Sum; System; Tail; Techniques; Technology; Therapeutic; Time; Transgenes; Transgenic Mice; Transgenic Organisms; Universities; Uterus; Vendor; Work; YAC Clone; blastocyst; cost; cost effective; cost efficient; embryo cell; embryo cryopreservation; embryo quality; embryonic stem cell; endonuclease; experience; experimental study; flexibility; homologous recombination; implantation; insertion/deletion mutation; mouse genome; mouse model; offspring; pathogen; pregnant; pup; research and development; sperm cell; sperm cryopreservation; success; targeted nucleases; transcription activator-like effector nucleases

PROJECT SUMMARY (GENETICALLY MODIFIED MOUSE CORE) The significant incidence of digestive, liver and pancreatic diseases in the American population demands continued exploration of a broad array of corresponding mechanistic pathways, pathophysiologic sequelae, and potential therapeutic approaches. In many cases these investigations can only be accomplished, or can be accomplished most efficiently and relevantly, using model systems established in intact animals. The use of mouse models in these pursuits is now well established for their power, feasibility, flexibility, and enormous potential. Creation of such models, by targeted alterations of the mouse genome, is an essential component of an overall research effort in understanding normal functions and pathologic perturbations in the digestive tract, liver, and pancreas. The Genetically Modified Mouse Core (GMMC) provides investigators of the University of Pennsylvania Center for the Molecular Study of Digestive and Liver Diseases (CMSDLD) with the ability to carry out these technologically-demanding studies in a cost effective and efficient manner and enhance the rigor and relevance of these approaches. The GMMC has a dedicated and highly skilled staff that applies state-of-the-art equipment and techniques, and the facility consists of a microinjection suite, an adjacent dedicated cage room, and an off-site and highly secure cryopreservation storage facility. Major services available to CMSDLD investigators include the generation of transgenic mice by DNA pronuclear injection, creation of chimeric mice by ES cell injection into blastocysts, and direct genome mutation, editing, and modification via the use of targeted endonuclease (TALEN and Crispr-CAS technologies); these are complemented by the genotyping of founder mice, assisted (in vitro) fertilization, cryopreservation, long-term cryostorage, and shipping of frozen embryos or sperm to/from other facilities. To provide these services, the GMMC utilizes multiple microinjection platforms, laser-assisted technologies, state-of-the-art cryopreservation approaches, and highly efficient line re-derivation pipelines. All functions, from ordering services, to following workflow, to storing and sending out lines, are on-line and can be monitored in real-time. These efforts by the GMMC contribute substantially to the overall productivity of CMSDLD investigators and enhance the rigor and relevance of their studies to the mechanisms of digestive, liver, and pancreatic diseases in physiologically- intact mammalian systems. Moreover, the GMMC enhances interactions and collaborations for CMSDLD investigators and lowers the technical and financial barriers that would otherwise impede the application of these approaches for individual investigators. Thus the Genetically Modified Mouse Core contributes greatly to the research efforts of the CMSDLD.

项目总结（转基因小鼠核心） 美国人口中消化道、肝脏和胰腺疾病的显著发病率要求 继续探索广泛的相应机制途径，病理生理后遗症， 和潜在的治疗方法。在许多情况下，这些调查只能完成，或可以 使用在完整动物中建立的模型系统，可以最有效和最相关地完成。使用 小鼠模型在这些追求中的地位已经确立，因为它们的力量、可行性、灵活性和巨大的 潜力通过有针对性地改变小鼠基因组来创建此类模型是 全面研究了解消化道的正常功能和病理扰动， 肝脏和胰腺转基因小鼠核心（GMMC）提供了研究人员的大学 宾夕法尼亚消化和肝脏疾病分子研究中心（CMSDLD）有能力 以具成本效益和有效率的方式进行这些技术要求高的研究，并加强 这些方法的严谨性和相关性。GMMC拥有一支敬业且技术精湛的员工队伍， 最先进的设备和技术，该设施包括显微注射套件、相邻的 专门的笼室，以及一个场外和高度安全的低温保存储存设施。主要服务 CMSDLD研究者可用的方法包括通过DNA原核注射产生转基因小鼠， 通过将ES细胞注射到胚泡中来产生嵌合小鼠，以及直接基因组突变、编辑和 通过使用靶向核酸内切酶（TALEN和Crispr-CAS技术）进行修饰;这些是 辅之以创始小鼠的基因分型、辅助（体外）受精、冷冻保存、长期 冷冻保存和冷冻胚胎或精子往返于其他设施的运输。为了提供这些服务， GMMC利用多种显微注射平台、激光辅助技术、最先进的冷冻保存技术 方法和高效的线条重新推导流水线。所有功能，从订购服务到以下 工作流程，存储和发送线，是在线的，可以实时监控。的这些努力 GMMC大大提高了CMSDLD研究者的整体生产力，并提高了研究的严谨性和 他们的研究与消化，肝脏和胰腺疾病的生理机制的相关性- 完整的哺乳动物系统此外，GMMC增强了CMSDLD的互动和协作 调查人员，并降低技术和财政障碍，否则将阻碍应用 这些方法对于个体研究者来说。因此，转基因小鼠核心大大有助于 CMSDLD的研究工作。