Inducible site-specific recombination in myelinating cells
Inducible site-specific recombination in myelinating cells
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DOI：
10.1002/gene.10154
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发表时间：
2003-01-01
期刊：
影响因子：
1.5
通讯作者：
Popko, B
中科院分区：
文献类型：
--
作者：
Doerflinger, NH;Macklin, WB;Popko, B
为了探索由髓鞘形成细胞表达的基因的功能，我们已经开发了一个模型系统，该系统允许在少突胶质细胞和许旺细胞中诱导消融预定基因。Cre/ IoxP重组系统提供了在小鼠中产生组织特异性体细胞突变的机会。我们已经使用Cre重组酶和人雌激素受体（CreER（T））的突变配体结合结构域之间的融合蛋白来获得可诱导的位点特异性重组。CreER（T）表达置于髓磷脂蛋白脂质蛋白（PLP）基因的调节序列的转录控制下，该基因在少突胶质细胞中大量表达，在许旺细胞中表达程度较低。CreER（T）融合蛋白易位至细胞核，并介导注射合成类固醇他莫昔芬的PLP/CreER（T）小鼠的髓鞘形成细胞中LacZ报告基因的重组。在未处理的动物中，CreER（T）保持在细胞质中，并且没有重组的证据。PLP/CreER（T）动物在阐明和区分特定基因在髓鞘形成和维持中的功能以及在分析病理条件下成熟少突胶质细胞功能方面应该是非常有用的。(C)2002 Wiley-Liss，Inc.
To explore the function of genes expressed by myelinating cells we have developed a model system that allows for the inducible ablation of predetermined genes in oligodendrocytes and Schwann cells. The Cre/ IoxP recombination system provides the opportunity to generate tissue-specific somatic mutations in mice. We have used a fusion protein between the Cre recombinase and a mutated ligand-binding domain of the human estrogen receptor (CreER(T)) to obtain inducible, site-specific recombination. CreER(T) expression was placed under the transcriptional control of the regulatory sequences of the myelin proteolipid protein (PLP) gene, which is abundantly expressed in oligodendrocytes and to a lesser extent in Schwann cells. The CreER(T) fusion protein translocated to the nucleus and mediated the recombination of a LacZ reporter transgene in myelinating cells of PLP/CreER(T) mice injected with the synthetic steroid tamoxifen. In untreated animals CreER(T) remained cytoplasmic, and there was no evidence of recombination. The PLP/CreER(T) animals should be very useful in elucidating and distinguishing a particular gene's function in the formation and maintenance of the myelin sheath and in analyzing mature oligodendrocyte function in pathological conditions. (C) 2002 Wiley-Liss, Inc.