Gene Targeting & Transgenic Mouse

基因打靶

基本信息

批准号：
8582094
负责人：
Israel DAVID GOLDMAN
金额：
$ 11.14万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
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/8582094
关键词：
Ablation Albert Einstein Cancer Center Alleles Animals Birth Blastocyst Transfer Cancer Center Cancer Center Support Grant Caring Cell Culture Techniques Cells Chimera organism Cloning Collaborations Complex Consult DNA DNA Sequence 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 Generations Genes Genetic Genetic Recombination Genomics 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 Mouse Knock-out Knockout Mice Laboratories Lentivirus 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 Ovarian 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 System Tamoxifen Techniques Technology Testing Tetracyclines Training Transgenes Transgenic Mice Transgenic Organisms Translational Research Transplantation 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 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（参见说明）：基因打靶和转基因设施成立于 20 多年前，旨在协助 AECC 研究人员建立人类疾病的转基因小鼠模型。直到最近，基因打靶和转基因组件仍作为独立的共享资源运行，在上次 CCSG 审查中均被评为优秀。在搬迁到价格中心最先进的新实验室空间后，这些设施被合并为一个基因靶向和转基因共享资源。该设施的转基因部分通过原核注射或慢病毒感染将DNA序列（例如常规质粒载体或BAC克隆）引入种系，继续高效率（-100%成功率）产生转基因小鼠品系受精卵母细胞。对于每个项目，设施主管都会咨询各个研究人员，并就转基因构建体设计以及提供质粒和序列盒以确保适合在小鼠中表达提供建议。通常有 20-25 名研究人员使用该服务，每年注射大约 100 个构建体。基因靶向组件通过在胚胎干 (ES) 细胞中使用基因靶向方法并将这些变化引入小鼠种系，继续提供小鼠基因组修饰服务。基因打靶服务包括生成传统的基因敲除小鼠品系、基因敲入小鼠品系以及具有用于基因时间和空间消融的条件等位基因的小鼠品系。该设施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 研究人员提供快速且经济有效地生成转基因小鼠品系的完整服务。服务包括设计和生成简单和复杂的基因打靶载体，电穿孔和筛选各种遗传背景（129、B6和129/06）的胚胎干细胞系，以及通过囊胚注射产生嵌合小鼠（通过基因打靶进行）设施）。基因修饰服务还开发了用于高通量生成靶向载体和筛选程序的新技术和程序，并于 2012 年初开始全面运营。