Improvement and application of cerebral cortex-specific gene manipulation system

大脑皮层特异性基因操作系统的改进及应用

• 批准号: 13680917
• 负责人: IWASATO Takuji
• 金额: $ 2.3万
• 依托单位: RIKEN (The Institute of Physical and Chemical Research)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13680917/
• 关键词: Emx1; conditional knockout; reverse genetics; transgenic mice; PAC; neuroscience; developmental biology; Cre; Emx1; 神経科学; プロモーター; 条件的遺伝子ノックアウト; 免疫組織化学; loxP

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 3 transgenic mouse lines. In all 3 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 3 lines in adulthood. Thus, 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.

将基因表达或破坏限制在大脑特定区域的能力将增强对大脑发育和功能的分子基础的理解。为此，脑区域限制性启动子是必不可少的。在这里，我们报告的分离的DNA片段包含Emx 1基因启动子，这是负责背端脑特异性表达。将Cre重组酶基因插入小鼠PAC（P1衍生的人工染色体）Emx 1-基因座克隆（PAC-Emx 1#1克隆）中，并用于产生3个转基因小鼠系。在所有3个系，特别是Tg 3，Cre介导的重组高度限制于Emx 1表达细胞系，从胚胎期到成年期。免疫组织化学分析表明，Cre蛋白表达在背侧端脑在所有3行在成年。因此，PAC-Emx 1 #1克隆基本上含有Emx 1基因表达所需的所有调控元件。我们的研究结果表明，Emx 1-Cre Tg 3小鼠和PAC-Emx 1 #1克隆构成了背侧端脑特异性基因操作的强大工具。

项目成果

岩里琢治(分担執筆): "「神経活動依存的メカニズムによる脳回路発達」みる見るわかる脳・神経科学入門講座(下)"羊土社. 15 (2002)

Takuji Iwasato（合著者）：“‘通过神经活动依赖机制实现脑回路发展’ - 易于理解的大脑和神经科学入门课程（第 2 部分）”Yodosha。

Inoue, H., Tsukita, K., Iwasato.T., et al.: "The crucial role of caspase in the disease progression of a transgenic ALS mouse model."EMBO J.. 22. 6665-6674 (2003)

Inoue, H.、Tsukita, K.、Iwasato.T. 等人：“半胱天冬酶在转基因 ALS 小鼠模型疾病进展中的关键作用。”EMBO J.. 22. 6665-6674 (2003)

岩里琢治, 糸原重美: "マウス遺伝学による神経回路網精緻化の研究"生体の科学. 54. 6665-6674 (2003)

Takuji Iwasato、Shigemi Itohara：“利用小鼠遗传学详细阐述神经网络的研究”Bioscience 54. 6665-6674 (2003)。

Iwasato, T., Nomura, R., Ando, R., Ikeda, T., Tanaka, M, Itohara, S.: "Dorsal Telencephalon-Specific Expression of Cre Recombinase in PAC Transgenic Mice."Genesis. 38. 130-138 (2004)

Iwasato, T.、Nomura, R.、Ando, R.、Ikeda, T.、Tanaka, M、Itohara, S.：“PAC 转基因小鼠中 Cre 重组酶的背侧端脑特异性表达”。

Datwani, A., Iwasato, T., Itohara, S., Erzurumlu, R.S.: "MDA receptor-dependent pattern transfer from afferents to postsynaptic cells and dendritic differentiation in the barrel cortex."Mol.Cell.Neurosci.. 21. 477-492 (2002)

Datwani, A.、Iwasato, T.、Itohara, S.、Erzurumlu, R.S.：“MDA 受体依赖性模式从传入细胞转移到突触后细胞以及桶状皮质中的树突分化。”Mol.Cell.Neurosci.. 21. 477