C10155: Reporters for Visualizing Signaling Pathways in Living CellsValue Proposition: Protein kinases and phosphatases are crucial components of intracellular signaling pathways which transmit signals by phosphorylation of downstream targets, altering their function. Efficient signal transduction requires precise kinase regulation within specific biological contexts, making tools that allow study of their dynamics in situ critical for understanding kinase function. JHU scientists have developed genetically encodable reporters to monitor spatio-temporal dynamics of specific enzymes in living cells.
• A reporter system – Bimolecular Indicator Reporting (BindR) useful in generating biosensors of a wide range of target kinases.
• A FRET-based reporter that provides a flexible, reliable and quantifiable measurement of Jun N-terminal kinase (JNK) activity in living cells.
• A FRET-based reporter that provides a flexible, reliable and quantifiable measurement of Extracellular Signal-Regulated Kinase (ERK) activity in living cells.
• A reporter which provides a unique tool to study Akt signaling in living cells where Akt activity can be monitored in real time.
• A FRET-based reporter to detect phosphatase activity in living cells that directly reports Protein Kinase A activity
• A FRET-based reporter to detect phosphatase activity in living cells that directly reports Protein Kinase C activity
• A FRET-based reporter to detect phosphatase activity in living cells that directly reports calcineurin activity.
Technical Details:
Genetically encoded reporters based on fluorescence resonance energy transfer (FRET) have become powerful tools for monitoring cellular events in living systems. There is a continuing need for sensitive enzyme reporters, particularly genetically encoded reporters, which can be used to measure enzyme activities with high spatial and temporal resolution in vivo.
Looking for Partners:
This technology provides genetically encoded reporters that can be used to measure intracellular enzymatic activity in living cells. The reporters are additionally useful for high-throughput drug screening and to monitor enzymatic activity in various disease states.
• Increased sensitivity and specificity of the FRET-based reporters allows for monitoring of signaling activity in living cells in real-time
• Reliable and quantifiable emission spectra allow development of high throughput assays
• Provides tunable (flexible) reporters with different spectral properties for the development of assays with greater efficiencies and sensitivity
• May be targeted to different subcelluar locations providing high spatial and temporal resolution of enzyme activity
