Iwasato Takuji, Nomura Ryochi, Ando Reiko, Ikeda Toshio, Tanaka Mika, Itohara Shigeyoshi
PRESTO, Japan Science and Technology Agency, Saitama, Japan.
Genesis. 2004 Mar;38(3):130-8. doi: 10.1002/gene.20009.
The ability to restrict gene expression or disruption to specific regions of the brain would enhance understanding of the molecular basis for brain development and function. For this purpose, brain region-restricted promoters are essential. Here we report the isolation of a DNA fragment containing the Emx1 gene promoter, which is responsible for dorsal telencephalon-specific expression. The Cre recombinase gene was inserted into a mouse PAC (P1-derived artificial chromosome) Emx1-locus clone (PAC-Emx1#1 clone) and utilized to generate three transgenic mouse lines. In all three lines, especially Tg3, Cre-mediated recombination was highly restricted to Emx1-expressing cell lineages, from embryonic stages to adulthood. Immunohistochemical analyses showed that Cre protein is expressed in the dorsal telencephalon in all three lines in adulthood. Thus, the PAC-Emx1#1 clone contains essentially all regulatory elements necessary for Emx1 gene expression. Our results suggest that Emx1-Cre Tg3 mice and the PAC-Emx1#1 clone constitute powerful tools for dorsal telencephalon-specific gene manipulation.
将基因表达限制或破坏在大脑特定区域的能力，将有助于增进对大脑发育和功能分子基础的理解。为此，脑区特异性启动子至关重要。在此，我们报告了一个包含Emx1基因启动子的DNA片段的分离，该启动子负责端脑背侧特异性表达。将Cre重组酶基因插入小鼠PAC（P1衍生人工染色体）Emx1基因座克隆（PAC-Emx1#1克隆），并用于生成三个转基因小鼠品系。在所有三个品系中，尤其是Tg3，从胚胎期到成年期，Cre介导的重组高度局限于表达Emx1的细胞谱系。免疫组织化学分析表明，成年期所有三个品系的端脑背侧均表达Cre蛋白。因此，PAC-Emx1#1克隆基本上包含了Emx1基因表达所需的所有调控元件。我们的结果表明，Emx1-Cre Tg3小鼠和PAC-Emx1#1克隆构成了用于端脑背侧特异性基因操作的强大工具。
