Dual-Color Luciferase Mouse Directly Demonstrates Coupled Expression of Two Clock Genes
Dual-Color Luciferase Mouse Directly Demonstrates Coupled Expression of Two Clock Genes
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DOI：
10.1021/bi100545h
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发表时间：
2010-09-21
期刊：
影响因子：
2.9
通讯作者：
Nakajima, Yoshihiro
中科院分区：
文献类型：
--
作者：
Noguchi, Takako;Michihata, Tomoko;Nakajima, Yoshihiro
我们已经建立了一个双色转基因小鼠，同时报告两个时钟基因，Bmal1和Per2，在一个单一的组织中的表达。这两个基因的表达监测与绿色和红色发光的甲虫酶与一个单一的发光底物。Bmal1和Per2的反相振荡，其内源性mRNA的配置文件一致，清楚地监测视交叉上核（SCN），主昼夜节律起搏器，并在外周组织，表明该系统允许长期的，定量的，同时监测两个基因的表达。我们还表明，虽然Bmal1和Per2在每个器官中的表达模式是严格反相的，但记录的昼夜节律阶段和两个基因的周期在器官之间变化。在SCN中的两个基因的表达中的相移，由培养基的变化引起的，也以类似的方式发生。因此，这种双色荧光素酶小鼠允许非侵入性和连续监测两个时钟基因的偶联表达。该系统提供了一种简单的技术，可以解开体内两个基因之间复杂的相互作用。
We have established a dual-color transgenic mouse that simultaneously reports the expression of two clock genes, Bmal1 and Per2, in a single tissue. The expression of the two genes is monitored with green- and red-emitting beetle luciferases with a single luminescent substrate. Antiphasic oscillations of Bmal1 and Per2, consistent with their endogenous mRNA profiles, were clearly monitored in the suprachiasmatic nucleus (SCN), the master circadian pacemaker, and in the peripheral tissues, demonstrating that the system allows the long-term, quantitative, and simultaneous monitoring of the expression of the two genes. We also showed that although the expression patterns of Bmal1 and Per2 in each organ are strictly antiphasic, the recorded circadian phases and periods of both genes varied between organs. The phase shifts in the expression of both genes in the SCN, induced by a change of medium, also occurred in a similar manner. Therefore, this dual-color luciferase mouse allows noninvasive and continuous monitoring of the coupled expression of two clock genes. This system provides a simple technique with which to unravel the complex interactions of two genes in the body.