Efficient recombination in diverse tissues by a tamoxifen-inducible form of Cre: A tool for temporally regulated gene activation/inactivation in the mouse
Efficient recombination in diverse tissues by a tamoxifen-inducible form of Cre: A tool for temporally regulated gene activation/inactivation in the mouse
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DOI：
10.1006/dbio.2002.0597
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发表时间：
2002-04-15
影响因子：
2.7
通讯作者：
McMahon, AP
中科院分区：
文献类型：
--
作者：
Hayashi, S;McMahon, AP
近年来，来自噬菌体P1的Cre整合酶已成为小鼠条件基因激活和/或失活的重要工具。在早期的报告中，我们描述了Cre和雌激素受体配体结合域突变形式（Cre- er (TM)）之间的融合蛋白，该融合蛋白使Cre活性他莫昔芬（TM）可诱导，允许在哺乳动物子宫内神经管中有条件地修饰基因活性。在目前的工作中，我们已经产生了一种转基因小鼠系，其中Cre-ER（TM）无处不在地表达，允许在胚胎和成年阶段的小鼠不同组织中暂时调节cre介导的重组。我们证明，在性交后8.5天，将TM单次腹腔注射到怀孕小鼠体内，可在注射后6小时内检测到发育中的胚胎中的重组，并在注射后24小时内有效地重组了所有三个胚层衍生物中的报告基因。此外，通过改变注射TM的剂量，可以控制胚胎中发生重组事件的细胞百分比。在成年小鼠的多种组织（包括中枢神经系统）和转基因小鼠系的培养细胞中，TM诱导的剂量依赖性切除也是可能的。这种可诱导的Cre系统将成为一种广泛有用的工具，用于调节小鼠胚胎，成人和培养系统中的基因活性，其中时间控制是所有重要的考虑因素。(C) 2002 Elsevier Science （USA）。
In recent years, the Cre integrase from bacteriophage P1 has become an essential tool for conditional gene activation and/or inactivation in mouse. In an earlier report, we described a fusion protein between Cre and a mutated form of the ligand binding domain of the estrogen receptor (Cre-ER(TM)) that renders Cre activity tamoxifen (TM) inducible, allowing for conditional modification of gene activity in the mammalian neural tube in utero. In the current work, we have generated a transgenic mouse line in which Cre-ER(TM) is ubiquitously expressed to permit temporally regulated Cre-mediated recombination in diverse tissues of the mouse at embryonic and adult stages. We demonstrate that a single, intraperitoneal injection of TM into a pregnant mouse at 8.5 days postcoitum leads to detectable recombination in the developing embryo within 6 h of injection and efficient recombination of a reporter gene in derivatives of all three germ layers within 24 h of injection. In addition, by varying the dose of TM injected, the percentage of cells undergoing a recombination event in the embryo can be controlled. Dose-dependent excision induced by TM was also possible in diverse tissues in the adult mouse, including the central nervous system, and in cultured cells derived from the transgenic mouse line. This inducible Cre system will be a broadly useful tool to modulate gene activity in mouse embryos, adults, and culture systems where temporal control is all important consideration. (C) 2002 Elsevier Science (USA).