BAC Transgenic Mice Encoding Tau Green Through the Nurr1

BAC 转基因小鼠通过 Nurr1 编码 Tau Green

• 批准号: 6987935
• 负责人: Cristina Backman
• 金额: --
• 依托单位: NATIONAL INSTITUTE ON DRUG ABUSE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6987935
• 关键词: artificial chromosomes; biotechnology; brain mapping; dopamine; gene expression; genetic regulatory element; genetically modified animals; laboratory mouse; mesencephalon; nucleic acid sequence; olfactory lobe; site directed mutagenesis; tau proteins

The recent development of methods for engineering bacterial artificial chromosomes (BACs), and for the efficient production of BAC transgenic mice, has simplified the design of in vivo approaches for the analysis of gene expression and function in the brain. BAC transgenic mice carrying an exogenous 150 kb DNA sequence containing the Nurr1 promoter directly followed by the Tau green (TG) gene have been generated. The exogenous DNA, was randomly integrated in the genome when injected into oocytes. As the carrying capacity of BACs is several hundred kilobases, we were able to insert a 150 Kb DNA fragment overlapping the Nurr1 gene (as compared to other carrying vectors allowing only 10-15 kb inserts). In these transgenic mice, Tau green expression was expressed only in the olfactory bulb, and absent in other brain areas known to express Nurr1. This data suggests that the BAC (containing the Nurr1 promoter) used in this study is lacking some of the regulatory elements necessary for driving TG expression in brain regions other than the olfactory bulb. We are currently developing additional BAC transgenic mice, for mapping/visualizing the midbrain dopaminergic system. These mice will be used in our laboratory to conduct several anatomical/functional studies in which mapping/visualization of the Nurr1/midbrain dopaminergic system is necessary.

工程细菌人造染色体（BAC）以及有效产生BAC转基因小鼠的方法的最新发展已简化了体内方法的设计，用于分析大脑中基因表达和功能。已经产生了含有外源的150 kb DNA序列的BAC转基因小鼠，该序列含有Nurr1启动子，然后直接产生了Tau绿（TG）基因。当注射到卵母细胞中时，外源性DNA在基因组中随机整合。由于BAC的承载能力为几百千碱基，因此我们能够插入150 kb的DNA片段重叠的Nurr1基因（与其他仅允许10-15 kb插入物的载体相比）。 在这些转基因小鼠中，tau绿色表达仅在嗅球中表达，并且在已知表达Nurr1的其他大脑区域不存在。该数据表明，本研究中使用的BAC（包含Nurr1启动子）缺乏在嗅球以外的其他大脑区域驱动TG表达所需的调节元素。我们目前正在开发其他BAC转基因小鼠，用于绘制/可视化中脑多巴胺能系统。这些小鼠将在我们的实验室中用于进行几项解剖/功能研究，其中必须对Nurr1/中脑多巴胺能系统进行映射/可视化。