NanoLuc and GloSensor Technologies Enabling Kinetic Analysis of Signaling Pathways in iCell Cardiomyocytes Poster
Part # PS195
Jennifer Wilkinson, Jeff Kelly, Richard Somberg, Chris Eggers, Brock Binkowski, Jim Hartnett, Frank Fan, Keith Wood, Mei Cong and Matthew Robers
Promega Corporation; 2800 Wood Hollow Road, Madison WI 53711
Although traditional reporter gene assays have proven powerful tools to interrogate cell signaling pathways, transcriptional responses generally occur at distal signaling endpoints, limiting their suitability for real-time pathway analysis. Consequently, alternate methods that measure signaling responses at upstream pathway nodes are desirable as complements to transcriptional reporters. We have applied a new set of ultrasensitive bioluminescent reporter technologies ideally suited for these applications. First, NanoLuc® technology is ideally suited as a protein fusion tag for measuring changes in protein lifetime following induction of stress response signaling. Via genetic attachment of NanoLuc® luciferase to various transcription factors, induction of ROS and hypoxia responses can be measured. GloSensor™ technology exploits analyte-sensitive forms of permuted firefly luciferase for the purposes of quantifying intracellular cAMP levels upon induction of G-protein coupled receptors. The sensitivity of these reporters make them well-suited for applications in iCell® Cardiomyocytes using simple transfection methods.
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