Core--Mouse

小鼠的基因敲除对于确定它们编码的蛋白质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，那么特定基因产物在成人体内的功能是什么。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。这个核心的目标是敲除成人大脑中的CPEB基因，特别是海马体的CA1区域。为此，胚胎干细胞中的同源重组将被用于在CPEB基因的内含子中插入loxP位点。含有这种重组基因的小鼠将拥有正常的CPEB基因和蛋白质。然而，当这些小鼠在CaMKII启动子的调控下与其他含有Cre重组酶转基因的小鼠交配时，成年小鼠将在海马区CA1区的CPEB基因发生重组事件。这些小鼠实际上将是CPEB的零。这个核心将构建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，并执行所有的基因分型。核心还将保留几个品系的小鼠，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。在这些小鼠身上获得的结果将对包括学习和记忆在内的大脑功能产生重要影响。

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.