CORE--TRANSGENIC MICE

该计划项目赠款中的大多数项目将严重依赖转基因小鼠，其中一些已经生成，另一些需要生成。在过去的五年里，这个核心的存在极大地促进了由Rob Edwards、David哥本哈根、Louis Reichardt和Michael Stryker领导的实验室利用这项强大技术的能力。在接下来的五年里，我们预计这一核心的使用范围将扩大，几乎可以肯定的是，我们新项目团队的所有成员都将使用这一核心。核心的存在使我们能够聘请一名在准备转基因小鼠方面拥有超过15年相关经验的高级专家，以及一名能够管理小鼠群体和执行常规基因分型等的初级技术人员。资历较浅的技术人员 在帮助实验室进入这一领域方面特别有价值。所有实验室都将继续受益于转基因核心，有一个人致力于很好地完成某些在产生这种动物方面至关重要的任务。为了帮助培养和筛选胚胎干细胞，为转基因动物提供有针对性的突变，并帮助培训博士后研究员和研究生这项重要技术，我们建议维持一个中等规模和配备人员的核心设施，包括胚胎干细胞细胞培养设施、注射单细胞胚胎和囊胚的设备，以及一个小鼠群体，其中包括可生育和输精管切除的雄性和可生育雌性动物，用作单细胞胚胎或囊胚捐赠者和假孕接受者。赖哈特实验室将在首席调查员的全面指导下，为建立和管理这一设施提供监督和专业知识。

The majority of projects in this program project grant will rely heavily on transgenic mice, some of which have already been generated and others of which need to be generated. During the past five years, existence of this core has greatly facilitated the ability of the laboratories directed by Rob Edwards, David Copenhagen, Louis Reichardt and Michael Stryker to exploit this powerful technology. During the next five years, we anticipate expanded use of this core that will almost certainly be used by all members of our new program project team. The existence of a core has made it possible to hire a senior specialist with more than 15 years of relevant experience in preparing transgenic mice plus a junior technician able to manage the mouse colonies and perform routine genotyping, etc. The former person has helped our laboratories generate more than a dozen transgenic lines and half a dozen knockout strains. The more junior technician has been particularly valuable in helping the labs entering this field. All laboratories will continue to benefit from a transgenic core with a person dedicated to performing well certain tasks of critical importance in generating such animals. To help culture and screen ES cells, provide transgenic animals with targeted mutations and provide help in training postdoctoral fellows and graduate students in this important technology, we propose to maintain a modestly-sized and staffed core facility with a cell culture facility for embryonic stem cells, equipment for injection of single cell embryos and blastocysts, and a mouse colony with fertile and vasectomized males and fertile females for use as single cell embryo or blastocyst donors and pseudopregnant recipients. The Reichardt laboratory will provide the supervision and expertise for setting up and managing this facility under the overall guidance of the Principal Investigator.