Multiplexed Reporter Gene Assays: Monitoring the Cell Viability and the Compound Kinetics on Luciferase Activity
Multiplexed Reporter Gene Assays: Monitoring the Cell Viability and the Compound Kinetics on Luciferase Activity
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DOI：
10.1177/1087057111407768
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发表时间：
2011-08-01
影响因子：
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通讯作者：
Parker, Christian N.
中科院分区：
文献类型：
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作者：
Didiot, Marie-Cecile;Serafini, Sergio;Parker, Christian N.
具有多个读数的高通量筛选分析使人们能够监测多个分析参数。通过尽可能多地获取有关潜在生物学的信息，可以提高对真正活性的检测。本报告描述了一个扩展到标准荧光素酶报告基因测定，使多个参数从每个样品监测。该报告描述了多路荧光素酶试验与正交读数监测细胞活力使用还原瑞祖啉。此外，本技术说明表明，通过在活细胞中使用荧光素底物，可以记录测定时间过程。这使得鉴定通过抑制荧光素酶起作用的非活性或非特异性化合物以及改变基因表达或细胞生长的化合物成为可能。（Journal of biomolmolecular Screening 2011;16:786-793）
High-throughput screening assays with multiple readouts enable one to monitor multiple assay parameters. By capturing as much information about the underlying biology as possible, the detection of true actives can be improved. This report describes an extension to standard luciferase reporter gene assays that enables multiple parameters to be monitored from each sample. The report describes multiplexing luciferase assays with an orthogonal readout monitoring cell viability using reduction of resazurin. In addition, this technical note shows that by using the luciferin substrate in live cells, an assay time course can be recorded. This enables the identification of nonactive or unspecific compounds that act by inhibiting luciferase, as well as compounds altering gene expression or cell growth. (Journal of Biomolecular Screening 2011;16:786-793)