Analysis of the Transcriptional Activity of Retinoic Acid-Related Orphan Receptors (RORs) and Inhibition by Inverse Agonists.

Analysis of the Transcriptional Activity of Retinoic Acid-Related Orphan Receptors (RORs) and Inhibition by Inverse Agonists.
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视黄酸相关孤儿受体 (ROR) 的转录活性分析和反向激动剂的抑制。

DOI:
10.1007/978-1-4939-9195-2_15
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发表时间:
2019
期刊:
Methods Mol. Biol.
影响因子:
--
通讯作者:
and Jetten A.M.
and Jetten A.M.
中科院分区:
--
文献类型:
--
作者:
Takeda Y.;Kang H.S.;and Jetten A.M.

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在这里,我们描述了几种测定来分析视黄酸相关孤儿受体(ROR)的转录活性和反向激动剂对其活性的影响。一种测定法测量反向激动剂对Tet-On细胞系统中ROR对荧光素酶报告基因的转录激活的影响。哺乳动物双杂交测定分析ROR配体结合结构域与共激活肽的相互作用。另外两个测定使用哺乳动物单杂交测定检查反向激动剂对在ROR靶基因IL 17的启动子控制下的荧光素酶报告基因的活化和对ROR介导的活化的影响。
Here, we describe several assays to analyze the transcriptional activity of retinoic acid-related orphan receptors (RORs) and the effect of inverse agonists on their activity. One assay measures the effect of an inverse agonist on the transcriptional activation of a luciferase reporter by RORs in a Tet-On cell system. A mammalian two-hybrid assay analyzes the interaction of the ROR ligand binding domain with a coactivator peptide. Two additional assays examine the effect of an inverse agonist on the activation of a luciferase reporter under control of the promoter of the ROR target gene,IL17, and on ROR-mediated activation using a mammalian monohybrid assay.
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