Cellular MicroRNA Sensors Based on Luciferase Reporters
Cellular MicroRNA Sensors Based on Luciferase Reporters
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DOI：
10.1007/978-1-62703-703-7_11
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发表时间：
2014-01-01
期刊：
影响因子：
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通讯作者：
Deiters, Alexander
中科院分区：
文献类型：
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作者：
Connelly, Colleen M.;Deiters, Alexander
最近，microRNA (miRNA) 已被证实与多种人类疾病有关，包括癌症和病毒感染。 miRNA 的小分子修饰剂可以代表新的治疗剂，并可用作阐明 miRNA 生物学作用的工具。为了识别 miRNA 的小分子修饰物，必须开发和优化针对特定 miRNA 的功能测定。在这里，我们报告了用于 miRNA miR-122 功能的荧光素酶报告基因测定的构建，以及可用于小分子 miR-122 抑制剂高通量筛选的稳定 Huh7 细胞系的开发。这里描述的步骤不仅适用于 Huh7 细胞和 miR-122，而且适用于几乎任何细胞系和 miRNA 组合。
Recently, microRNAs (miRNAs) have been linked to a variety of human diseases including cancer and viral infections. Small molecule modifiers of miRNAs could represent new therapeutic agents and be used as tools for elucidating the biological roles of miRNAs. In order to identify small molecule modifiers of miRNAs, functional assays for specific miRNAs must be developed and optimized. Here, we report the construction of a luciferase reporter assay for miRNA miR-122 function and the development of a stable Huh7 cell line that can be used for high-throughput screening of small molecule miR-122 inhibitors. The steps described here can be applied not only to Huh7 cells and miR-122 but also to virtually any cell line and miRNA combination.