Doxycycline-mediated quantitative and tissue-specific control of gene expression in transgenic mice
Doxycycline-mediated quantitative and tissue-specific control of gene expression in transgenic mice
复制标题
DOI：
10.1073/pnas.93.20.10933
复制
发表时间：
1996-10-01
影响因子：
11.1
通讯作者：
Bujard, H
中科院分区：
文献类型：
--
作者：
Kistner, A;Gossen, M;Bujard, H
将强力霉素（dox）作为特异性工程RNA聚合酶II启动子诱导剂的tet调控系统转移到转基因小鼠中。根据动物供水中的还原浓度，监测到严格的控制和广泛的调节范围，可达5个数量级。在一些器官中，给药还原可迅速诱导指示酶荧光素酶的合成，其活性在前4小时内上升几个数量级，在大多数器官中，诱导在24小时后完成。在tet调控系统中，发现了类似的调控潜力，其中还原可以阻止转录激活，将四环素控制的反激活子（tTA）的合成引导到肝脏，导致肝细胞中高度特异性的调控，在还原存在时，每个细胞检测到的荧光素酶分子少于一个，相比之下，在没有抗生素的情况下，观察到的荧光素酶基因的激活超过10(5)倍。这些结果表明，这两种四环素控制的转录激活系统都提供了基因开关，允许以组织特异性的方式对转基因小鼠的基因活性进行定量控制，因此，提出了产生新型条件突变体的可能性。
The tet regulatory system in which doxycycline (dox) acts as an inducer of specifically engineered RNA polymerase II promoters was transferred into transgenic mice. Tight control and a broad range of regulation spanning up to five orders of magnitude were monitored dependent on the dox concentration in the water supply of the animals, Administration of dox rapidly induces the synthesis of the indicator enzyme luciferase whose activity rises over several orders of magnitude within the first 4 h in some organs, Induction is complete after 24 h in most organs analyzed. A comparable regulatory potential was revealed with the tet regulatory system where dox prevents transcription activation, Directing the synthesis of the tetracycline-controlled transactivator (tTA) to the liver led to highly specific regulation in hepatocytes where, in presence of dox, less than one molecule of luciferase was detected per cell, By contrast, a more than 10(5)-fold activation of the luciferase gene was observed in the absence of the antibiotic. This regulation was homogeneous throughout but stringently restricted to hepatocytes, These results demonstrate that both tetracycline-controlled transcriptional activation systems provide genetic switches that permit the quantitative control of gene activities in transgenic mice in a tissue-specific manner and, thus, suggest possibilities for the generation of a novel type of conditional mutants.