CORE--ANIMAL RESOURCES

核心——动物资源

• 批准号: 6316686
• 负责人: ANN MARIE LAWLER
• 金额: $ 20.89万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-05-25 至 2000-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6316686
• 关键词: Downs syndrome; animal breeding; animal colony; animal population genetics; biomedical facility; chromosome 21; cryopreservation; disease /disorder model; embryo /fetus tissue /cell culture; embryonic stem cell; genetic strain; genetically modified animals; laboratory mouse; microinjections; polymerase chain reaction; southern blotting

The goal of the Animal Resources Core will continue to be to generate and maintain transgenic mouse lines for use in subprojects. Following the successful generation of a number of mouse lines during the previous granting period, the efforts of the Animal Resources Core will be shifting towards maintenance and breeding of existing transgenic mouse lines. The Core will continue to provide microinjection services for generating additional lines of mice to fulfill the goals of the research projects. This service will be primarily directed towards injections of genetically manipulated embryonic stem cell lines into blastocysts, but will also offer pro-nuclear injections of DNA when needed. Based on the predictions of the principal investigators, the Core expects to provide microinjection services for twelve to fifteen cell lines each year. The Core will be directing more of its efforts into freezing embryos from lines of transgenic mice in order to reduce animal care costs and to facilitate transfer of mice between institutions. During periods between microinjections, the breeding stocks and mice used for generating pseudopregnant females will be used for testing conditions for producing embryos for freezing and for testing frozen stocks of embryos. A CryoLogic CL 863 programmable controlled rate freezing unit has been purchased for use by Core C personnel and initial experiments to test freezing protocols have been successful. The primary function of the Core will be to maintain breeding stocks and produce study animals for the various research projects. The Core oversees breeding plans, performs DNA typing, maintains breeding records and coordinates with investigators for the production of animals in the required numbers and at the appropriate ages for experiments. The Core also maintains a line for FVB/N mice carrying a pSV40tkneo construct that is used to produce neomycin resistant embryonic fibroblasts for culture and selection of embryonic stem cells. One new function of the Core will be to maintain a colony of TS65Dn for use by investigators within the program at the Johns Hopkins School of Medicine. The TS65Dn colony was obtained from the laboratory of Dr. Muriel Davisson and is maintained according to the same breeding protocols. Genetic typing of the mice will be performed FISH analysis of blood from trails using a rapid and convenient protocol developed by Core D: Cytogenetics. All animals will be maintained in micro-isolator cages in a pathogen-fee facility at the Johns Hopkins School of Medicine. The Core has the ability to import lines of mice from external facilities as needed by performing embryo transfers with sentinel follow-up.

动物资源核心的目的将继续是生成和维护转基因小鼠线以用于子项目。在上一个授予期间成功产生了许多小鼠线，动物资源核心的努力将转移到现有转基因小鼠线的维持和繁殖。核心将继续提供微注射服务，以生成其他小鼠以实现研究项目的目标。该服务将主要针对将遗传操纵的胚胎干细胞系注射到胚泡中，但在需要时也将提供核核注射DNA。根据主要研究人员的预测，核心希望每年提供十二至15个细胞系的微注射服务。核心将把更多的努力引导到从转基因小鼠系列中冻结胚胎，以降低动物护理成本并促进机构之间的小鼠转移。在微型注射之间，用于产生假性雌性的繁殖量和小鼠将用于测试生产冻结胚胎和测试冷冻胚胎的胚胎的条件。 Cerologic CL 863可编程控制的速率冻结单元已被核心C人员使用，并且最初的实验测试冻结协议已成功。核心的主要功能是维持繁殖库存并为各种研究项目生产研究动物。核心监督育种计划，执行DNA键入，维持育种记录并与研究人员在所需数量和适当年龄的实验中生产动物的研究人员。该核心还维持了携带PSV40TKNEO构建体的FVB/N小鼠的线，该构建体用于产生耐新霉素的胚胎成纤维细胞，用于培养和选择胚胎干细胞。核心的一个新功能是维护约翰·霍普金斯医学院计划中的调查人员使用TS65DN的殖民地。 TS65DN菌落是从穆里尔·戴维森（Muriel Davisson）博士实验室获得的，并根据相同的育种方案进行维护。将使用Core D：细胞遗传学开发的快速方便的方案对小鼠的遗传分型进行血液分析。所有动物都将在约翰·霍普金斯医学院的病原体式设施中保持在微隔离笼中。核心具有通过使用Sentinel随访的胚胎转移来从外部设施中导入小鼠线的能力。