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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.

项目概述（见说明）：基因靶向和转基因设施成立于20多年前，旨在帮助AECC研究人员建立人类疾病的转基因小鼠模型。直到最近，基因靶向和转基因组件作为独立的共享资源运行，在上一次CCSG审查中每个组件都被评为优秀。在搬迁到Price中心新的最先进的实验室空间后，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继续以高效率（-100%成功率），通过将DNA序列，如常规质粒载体或BAC克隆通过原核注射或慢病毒感染受精卵母细胞导入种系，产生转基因小鼠品系。对于每个项目，设施主管与个别研究人员进行磋商，并就转基因构建设计提出建议，并提供质粒和序列磁带，以确保适合在小鼠中表达。通常20-25名调查员使用该服务，每年注入约100个构造。基因靶向组件通过在胚胎干细胞（ES）中使用基因靶向方法并将这些变化引入小鼠种系，继续为小鼠基因组的修饰提供服务。基因靶向服务包括产生常规敲除小鼠系、敲入小鼠系和具有条件等位基因的小鼠系，用于基因的时间和空间消融。该设备获得基因靶向小鼠的成功率高达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研究人员提供快速和经济高效地生成转基因小鼠系的完整服务。服务包括设计和生成简单和复杂的基因靶向载体，电穿孔和筛选不同遗传背景的胚胎干细胞系（129，B6和129/06），以及通过囊胚注射产生嵌合小鼠（由基因靶向设施执行）。基因修饰服务处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年初开始全面运作。