Core--Mouse

小鼠的基因敲除对于确定其编码蛋白在整个动物中的生物学功能非常有用。然而，由于许多基因在整个发育过程中在多种组织中表达，因此很难确定，例如，如果敲除是胚胎致死的，则难以确定特定基因产物在成体中的功能。这个问题可以通过使用Cre重组酶/loxP系统的组织特异性基因敲除来规避。该核心的目标是敲除成年大脑中的CPEB基因，特别是海马体的CA 1区域。为此，将使用ES细胞中的同源重组将loxP位点插入CPEB基因的内含子中。含有这种重组基因的小鼠将具有正常的CPEB基因和蛋白质。然而，当这些小鼠与其他小鼠交配含有Cre重组酶转基因的CaMKII启动子的调节下，成年人将经历在海马CA 1区的CPEB基因座的重组事件。这些小鼠将有效地对CPEB无效。该核心将构建用于CPEB重组的靶向载体，电穿孔和筛选ES细胞，注射囊胚，进行胚胎移植，并进行所有基因分型。核心还将维持几个品系的小鼠，因为它们与这些实验有关。这些小鼠将在分子、细胞、神经生理学和行为水平上进行分析。这些小鼠获得的结果将对大脑功能，包括学习和记忆，产生重要影响。

Gene knockouts in mice are enormously useful for determining the biological functions of their encoded proteins in the whole animal. However, because many genes are expressed in a multitude of tissues throughout development, it is difficult to determine, for example, the function a particular gene product in the adult if the knockout is embryonic lethal. This problem can be circumvented by tissue-specific gene knockouts using the Cre recombinase/loxP system. The goal of this Core is to knockout the CPEB gene in the adult brain, specifically the CA1 region of the hippocampus. To do this, homologous recombination in ES cells will be used to insert loxP sites into introns of the CPEB gene. Mice containing this recombined gene will have a normal CPEB gene and protein. However, when these mice are mated with other mice containing a Cre recombinase transgene under the regulation of the CaMKII promoter, adults will undergo a recombination event at the CPEB locus in the CA1 region of the hippocampus. These mice will effectively be null for CPEB. This Core will construct the targeting vector for CPEB recombination, electroporate and screen ES cells, inject blastocysts, perform the embryo transfer, and perform all the genotyping. The Core will also maintain several lines of mice as they pertain to these experiments. These mice will be analyzed at the molecular, cellular, neurophysiological, and behavioral levels. The results obtained with these mice will have important implications for brain function, including learning and memory.