Gene Targeting & Transgenic Mouse

基因打靶

基本信息

批准号：
9108263
负责人：
Israel DAVID GOLDMAN
金额：
$ 11.69万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE, INC
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-06-01 至 2018-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9108263
关键词：
Ablation Albert Einstein Cancer Center Alleles Animals Bacterial Artificial Chromosomes Birth Blastocyst Transfer Cancer Center Cancer Center Support Grant Caring Cell Culture Techniques Cells Chimera organism Cloning Collaborations Complex Consult DNA DNA Sequence DNA cassette Derivation procedure Developmental Biology ES Cell Line Educational process of instructing Electroporation Embryo Ensure FVB Mouse Fee-for-Service Plans Female Fertilization in Vitro Fibroblasts Freezing Gene Expression Gene Silencing Gene Targeting Gene-Modified Generations Genes Genetic Genetic Recombination Genomic DNA Germ Lines Goals Growth Health Herpes zoster disease Hormones Human Resources Inbred BALB C Mice Individual Infection Injection of therapeutic agent Institutes Instruction International Internet Knock-in Knock-in Mouse Knock-out Knockout Mice Laboratories Lentivirus Infections Maintenance Malignant Neoplasms Mammalian Oviducts Medicine Methods Microinjections Mission Modification Morbidity - disease rate Mothers Mouse Strains Mus Mutation NCI-Designated Cancer Center Online Systems Oocytes Ovulation Partner in relationship Phenotype Plasmid Cloning Vector Plasmids Preparation Price Procedures Production Protocols documentation Reagent Regulation Reproduction Research Personnel Resistance Resource Sharing Retrieval Safety Sampling Screening procedure Services Staging Subfamily lentivirinae System Tamoxifen Techniques Technology Testing Tetracyclines Training Transgenes Transgenic Mice Transgenic Organisms Translational Research Virus Weaning albino mouse base blastocyst college cost cost efficient design design and construction embryo stage 2 embryonic stem cell human disease interest male meetings mortality mouse genome mouse model new technology novel operation ovary transplantation pregnant promoter pup reagent testing research study screening sperm cell success transmission process vector zygote

项目摘要

PROJECT SUMMARY (See instructions): The Gene Targeting and Transgenic facility was founded over 20 years ago to assist AECC researchers in the generation of genetically modified mouse models of human disease. Until recently, the Gene Targeting and Transgenic components operated as independent Shared Resources, each being rated outstanding at the last CCSG review. Following relocation to new state-of-the-art laboratory space in the Price Center, the facilities were merged into one Gene Targeting and Transgenic Shared Resource. The Transgenic component of the facility continues to generate, with high efficiency (-100% success rate), transgenic mouse strains through the introduction of DNA sequences, such as regular plasmid vectors or BAC clones into the germ line by pronuclear injection or by lentivirus infection of fertilized oocytes. For each project, the facility supervisor consults with individual investigators and advises on transgene construct design, as well as making plasmids and sequence cassettes available to ensure suitable for expression in the mouse. Typically 20-25 investigators use this service with approximately 100 constructs injected per year. The Gene Targeting component continues to provide services for modification of the mouse genome through the use of gene targeting methods in embryonic stem (ES) cells and introduction of these changes into the mouse germline. Gene Targeting services include the generation of conventional knockout mouse lines, knock-in mouse lines and mouse lines with conditional alleles for the temporal and spatial ablation of genes. The facility has a high success rate (>95%) for obtaining gene targeted mouse. Finally, a new Gene Modification Service was implemented by the Albert Einstein College of Medicine to provide a complete service for the rapid and cost efficient generation of genetically modified mouse lines to all AECC investigators. Services include the design and generation of simple and complex gene targeting vectors, electroporation and screening of embryonic stem cell lines of various genetic backgrounds (129, B6 and 129/06) and the generation of chimeric mice by blastocyst injection (performed by the Gene Targeting facility). The Gene Modification Service has also developed new technologies and procedures for high-throughput generation of targeting vectors and screening procedures and began full operation in early 2012.

项目摘要（请参阅说明）：20年前建立了基因靶向和转基因设施，以帮助AECC研究人员生成转基因的人类疾病小鼠模型。直到最近，基因靶向和转基因成分作为独立共享资源运行，每个资源都在上次CCSG审查中被杰出。在搬迁到价格中心的新最先进的实验室空间后，设施被合并为一个基因靶向和转基因共享资源。该设施的转基因成分通过引入DNA序列（例如常规质粒载体或BAC克隆）通过前核注射或通过慢病毒感染受肥的卵母细胞引入种系，并以高效率（成功率为-100％的成功率）（成功率-100％的成功率）（成功率为-100％）。对于每个项目，设施主管都会与个别研究人员进行咨询，并就转基因构造设计提供建议，并制作质粒和序列盒，以确保适合在小鼠中表达。通常，20-25名调查人员每年使用大约100个构造，使用此服务。基因靶向成分通过使用基因靶向方法（ES）细胞（ES）细胞中的基因靶向方法以及将这些变化引入小鼠种系，以继续提供用于修饰小鼠基因组的服务。基因靶向服务包括传统的敲除小鼠线的产生，敲入小鼠线和带有条件等位基因的小鼠线的产生基因的时间和空间消融。该设施对于获得基因靶向小鼠的成功率很高（> 95％）。最后，阿尔伯特·爱因斯坦医学院实施了一项新的基因修饰服务，为所有AECC调查人员提供了快速，成本效率地生成转基因的小鼠系列的完整服务。服务包括靶向向量的简单和复杂基因的设计和生成，各种遗传背景的胚胎干细胞系的电穿孔和筛选（129，B6和129/06）以及通过胚泡注射（由基因靶向设备执行）的嵌合小鼠产生。基因修饰服务还开发了新的技术和程序，用于高通量生成靶向向量和筛选程序，并于2012年初开始全面运行。