Description
The GloResponse™ CRE- luc2P HEK293 Cell Line is a clonal derivative of Human Embryonic Kidney 293 cells. These cells contain a luciferase gene ( luc2P ) under the control of a minimal Herpes Simplex Virus (HSV) thymidine kinase promoter with multiple cAMP response elements (CREs). CRE is the DNA-binding sequence for the transcription factor CRE binding protein (CREB), which is responsible for regulation of a variety of biological functions including cell proliferation, circadian rhythms and memory. The GloResponse™ CRE- luc2P HEK293 Cell Line is designed for rapid and convenient analysis of any cellular response that results in modulation of CREB activities.
GPCRs regulate a wide-range of biological functions and are one of the most important target classes for drug discovery. GPCR signaling pathways can be categorized into three classes based on the G protein α-subunit involved: Gs, Gi/o and Gq. The GloResponse™ CRE- luc2P HEK293 Cell Line can be used to study and configure screening assays for Gs- and Gi/o-coupled GPCRs, which signal through cAMP and the cAMP Response Element (CRE).
The GloResponse™ Cell Lines incorporate the improvements developed for the pGL4 family of reporter vectors for enhanced performance. The destabilized luc2P luciferase reporter is used for improved responsiveness to transcriptional dynamics. The luc2P gene is codon optimized for enhanced expression in mammalian cells, and the pGL4 plasmid backbone was engineered to reduce background reporter expression. The result is a cell line with very high induction levels when the pathway of interest is activated.
A total of 10,000 GloResponse™ CRE- HEK293 cells per well were dispensed into each well of a 96-well plate, and twofold serial dilutions of forskolin were added to induce reporter gene expression. After 4 hours of induction in a tissue culture incubator, luciferase activity was measured using the Dual-Glo® Luciferase Assay System Reagent on the GloMax® 96 Microplate Luminometer (n = 4).
