Cook Erica, Hermes Jeffrey, Li Jing, Tudor Matthew
Lead Discovery and Optimization, Bristol Myers Squibb, Pennington, NJ, USA.
Screening and Translational Enzymology, Roche, Basel, Canton of Basel-Stadt, Switzerland.
Methods Mol Biol. 2018;1755:179-195. doi: 10.1007/978-1-4939-7724-6_13.
While luminescent reporter gene assays allow for a rapid and relatively interference free assessment of the activation state of a luminescent reporter, fluorescent reporters do not. They suffer from artifacts such as compound fluorescence and cellular debris which makes the assessment of whole well fluorescence signals difficult. However, the use of high-content screening allows for the isolation of individual cells, segmentation and thus enables the screener to utilize fluorescent reporters to assess the activation state of such a high-content reporter on a cell by cell level, thus minimizing artifacts. Here we discuss the use of such a high-content reporter that enables screening for compounds useful for HIV reactivation on Jurkat cells with high-content screening.
虽然发光报告基因检测能够快速且相对无干扰地评估发光报告基因的激活状态，但荧光报告基因检测则不然。它们会受到诸如化合物荧光和细胞碎片等假象的影响，这使得对整个孔的荧光信号进行评估变得困难。然而，使用高内涵筛选可以分离单个细胞、进行分割，从而使筛选者能够利用荧光报告基因在单个细胞水平上评估此类高内涵报告基因的激活状态，进而将假象降至最低。在此，我们讨论使用这样一种高内涵报告基因，它能够通过高内涵筛选在Jurkat细胞上筛选出对HIV重新激活有用的化合物。
