Conference Presentation: Dynamic Live-Cell Visualization, Quantification of Akt Activity Using a Genetically-Encoded, Fluorescent Kinase Translocation Reporter

Overview

Dysregulation of signal transduction pathways is associated with cancer initiation, progression and recurrence.  The PI3K/Akt signaling pathway has been extensively investigated as a therapeutic target due to its mechanistic association with several Hallmarks of Cancer.

Studying dynamic changes in kinase activity can be difficult, and assays to measure these changes in live cells within a physiologically relevant environment are lacking. Standard approaches to monitoring Akt kinase activity are limited to end-point assays, which lack the ability to monitor the effects of treatments over time.

Here, we demonstrate the utility of the Incucyte^® Kinase Akt Lentivirus Reagent, encoding a kinase translocation reporter based on a green fluorescent protein-tagged Akt substrate whose subcellular localization is phosphorylation-dependent, and a red fluorescent nuclear protein to denote the nuclear/cytoplasmic boundary. Quantification of treatment responses using the Incucyte^® Live Cell Analysis System showed concentration-dependent inhibition of Akt activity for all compounds with varying kinetic profiles over 24 hours.

Watch the full video to learn more and see the utility of the Incucyte^® Kinase Akt Lentivirus Reagent in providing valuable kinetic measurements of Akt activity using live cells within a physiologically relevant environment.