Inducible raptor and rictor Knockout Mouse Embryonic Fibroblasts
Inducible raptor and rictor Knockout Mouse Embryonic Fibroblasts
复制标题
DOI：
10.1007/978-1-61779-430-8_16
复制
发表时间：
2012-01-01
期刊：
影响因子：
--
通讯作者：
Hall, Michael N.
中科院分区：
文献类型：
--
作者：
Cybulski, Nadine;Zinzalla, Vittoria;Hall, Michael N.
哺乳动物雷帕霉素靶蛋白（mTOR）激酶在称为mTOR复合物1（mTORC 1）和mTORC 2的两种结构和功能不同的多蛋白复合物中起作用。免疫抑制剂和抗癌药物雷帕霉素通常用于基础研究，作为研究mTOR信号传导的工具。然而，雷帕霉素仅抑制且仅不完全抑制mTORC 1，并且没有mTORC 2特异性抑制剂可用。因此，要全面了解体内mTOR信号传导，包括两种复合物的功能，除了药理学抑制外，还需要遗传抑制。利用Cre/LoxP系统，我们产生了可诱导敲除小鼠胚胎成纤维细胞（MEF）缺陷的mTORC 1特异性组件raptor（iRapKO）或mTORC 2特异性组件rictor（iRicKO）。诱导敲除是重要的，因为mTOR复合物组分是必需的。诱导raptor或rictor敲除分别消除raptor或rictor表达，并损害相应的mTOR信号分支。所描述的敲除MEF是单独研究两种mTOR复合物的全部功能的有价值的工具。
The mammalian Target of Rapamycin (mTOR.) kinase functions within two structurally and functionally distinct multiprotein complexes termed mTOR complex 1 (mTORC1) and mTORC2. The immunosuppressant and anticancer drug rapamycin is commonly used in basic research as a tool to study mTOR signaling. However, rapamycin inhibits only, and only incompletely, mTORC1, and no mTORC2-specific inhibitor is available. Hence, a full understanding of mTOR signaling in vivo, including the function of both complexes, requires genetic inhibition in addition to pharmacological inhibition. Taking advantage of the Cre/LoxP system, we generated inducible knockout mouse embryonic fibroblasts (MEFs) deficient for either the mTORC1-specific component raptor (iRapKO) or the mTORC2-specific component rictor (iRicKO). Inducibility of the knockout was important because mTOR complex components are essential. Induction of either raptor or rictor knockout eliminated raptor or rictor expression, respectively, and impaired the corresponding mTOR signaling branch. The described knockout MEFs are a valuable tool to study the full function of the two mTOR complexes individually.