Core--Mouse Genetics

描述(由申请人提供)：很明显，毒理学超级基金在未来十年的研究进展将取决于小鼠作为研究基础和临床相关问题的实验模型。人类和小鼠基因组的高度保守性使得利用转基因和敲除基因技术建立人类毒理学模型的方法非常吸引人。与此同时，老鼠和人类在新陈代谢和对有毒化学物质的反应方面的独特差异使人类基因进入老鼠的可能性很大。这一核心为这个超级基金项目提供了最先进的小鼠基因组遗传修改技术。转基因小鼠携带新的或新的基因，以及缺乏感兴趣的特定基因或包含关键基因的修改版本的“敲除”或“敲入”小鼠。提供了条件表达式和针对Cre/loxP的敲除策略。核心提供广泛的技术和专业知识密集型服务，包括实验设计 这些技术包括：胚胎干细胞的同源重组；胚胎干细胞的Cre转导以切除loxP；将转基因的胚胎干细胞显微注射到囊胚中以产生敲除或敲入小鼠；原核注射转基因或细菌人工染色体以产生转基因小鼠；慢病毒载体的胚胎注射；胚胎的冷冻保存；以及体外受精和卵巢移植等生育干预措施。为具有特殊服务和高优先级的项目量身定做服务。这个核心是一个突出的例子，说明那些不能合理地期望在个人基础上开发这些技术的研究人员如何能够获得非凡的专门技术、训练有素的专门人员和昂贵的设备。这种转基因和敲除小鼠核心的可获得性将使这些项目能够利用一系列多学科最先进的技术在小鼠身上进行多功能、尖端的分子遗传学研究。

DESCRIPTION (provided by applicant): It is clear that the progress of toxicological Superfund research during the coming decade will depend upon the mouse as an experimental model to investigate both basic and clinically relevant questions. The high degree of conservation in the genomes of humans and mice makes the approach of using transgene and knock-out gene technology to create models for human toxicology extremely attractive. At the same time, unique differences in metabolism and response to toxic chemicals between mouse and human make the substitution of human genes into the mouse compelling. This core provides this Superfund projects with the most advanced technologies for genetic modification of the mouse genome. Transgenic mice carrying new or novel genes and 'Knock-out' or 'Knock-in' mice lacking specific genes of interest or containing modified version of key genes are created. Conditional expression and Cre/LoxP targeted knock-out strategies are provided. The core provides a wide array of technology- and expertise-intensive services including experimental design consultation, embryonic stem cell homologous recombination, Cre transfection of embryonic stem cells for LoxP excision, blastocyst microinjection of genetically altered embryonic stem cells into blastocysts to create knock-out or knock-in mice, pronuclear injection of transgenes or bacterial artificial chromosomes to create transgenic mice, embryo injecion of Lentivirus vectors, cryopreservation of embryos, and fertility interventions such as in vitro fertilization and ovary transplant. Services are tailored for the projects with special service and high priority. This core is an outstanding example of how extraordinarily specialized techniques, highly trained dedicated personnel, and expensive equipment can be accessed by researchers who could not reasonably expect to develop them on an individual basis. The availability of this Transgenic and Knock-out Mouse Core will enable the projects to conduct versatile, cutting-edge, molecular genetic research in the mouse with a battery of multidisciplinary state-of-the-art techniques.