Genetically encoded fluorescent biosensors for GPCR discovery

While fluorescent dye-based assays are powerful tools for high throughput screening in engineered cell-lines or cell lysates to show that signaling has occurred, genetically-encoded sensors that measure dynamic processes in physiologically-relevant cells provide a higher level of insight into how signals are regulated in both space and time. It is the regulation of signaling that controls cell function in both health and disease.

The fluorescent assays for cAMP, DAG, and PIP2 described on these pages are compatible with both GPCR drug discovery and basic research. Each sensor is packaged in a BacMam vector for efficient delivery to a wide variety of cell types including cardiomyocytes, neurons, fibroblasts, pancreatic islet cells, and osteoblasts as well as standard cell lines such as HEK293, CHO and NIH3T3.

BacMam as a delivery vector was originally described by Boyce and Bucher in 1996, In mammalian cells, this vector, based on a baculovirus, drives strong and exclusive expression of the gene encoding our fluorescent biosensor. The amount of virus introduced into a well can be easily adjusted, so you can control the transduction efficiency and expression levels (Kost et al, 2007).

How much time does it take?

Montana-Molecular_assay-steps-diagram_Jan-2016

How do I get started?

For more details about the benefits of a live cell assay, check out Tebu-Bio’s blog

Archives