High-Content Reporter Assays

Abstract

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.

A New Method for Screening Natural Products to Stimulate IFN-γ Production in Jurkat Human T Lymphocytes

“…In general, bioluminescent reporters, such as firefly luciferase, and fluorescent reporters like green fluorescent protein (GFP) and red fluorescent protein (RFP) are two efficiently and widely used reporter proteins. 26,27 Reporter gene assay is a powerful tool used in high-content screening assays for novel drug discovery and studying some intracellular signaling pathways of interest, as well as cytokines and their induced genes in vitro. 28 What is more, the application of fluorescencebased cytokine reporter mice has also been well studied in vivo.…”