LiveWare Cell Lines & LiveWare Plasmid Vectors

LiveWare® is a series of cells and expression vectors that have been successfully used in drug discovery¹ and screening environments to aid the study of G-protein-coupled receptors (GPCR) that do not conventionally couple through intracellular calcium. 

Pharmaceutical companies currently use multiple assay types in their routine screening operations. Recent years have seen an increased interest in functional assays for G-protein-coupled receptors.  The development of the FLIPR® Fluorometric Imaging Plate Reader System has become an industry standard system for screening GPCRs that signal through calcium flux.  However, over 60% of the known G-protein-coupled receptors signal through pathways other than those which lead to an increase in intracellular calcium (i.e., other than those which couple through G_q), and as genomics reveals more G-protein-coupled receptor targets this trend continues to increase.

LiveWare is based on a series of G-protein chimeras that have been developed by Bruce Conklin and colleagues at the Gladstone Institute, UCSF. LiveWare also includes the promiscuous G-protein, G_α16. The chimeras consist of the alpha subunit of a G_q-protein complex whose 5 carboxy-terminal amino acids have been replaced with those from one of the other G-proteins (either G_αs, G_αi, G_αo, or G_αz). These amino acids are responsible for the coupling of the receptor to its G-protein. Co-expression of these chimeras with specific non-G_q-coupled receptors may result in the generation of an intracellular calcium signal upon receptor stimulation.

¹Coward, P. et al. Proc. Natl. Acad. Sci. 95(1): 352-357 (1998).

Features

• LiveWare includes host cells and plasmids that contain chimeras to enable G_αs, G_αi, G_αo, or G_αz coupled GPCR receptors to signal through calcium mobilization and take advantage of the high throughput capabilities of FLIPR Calcium Assay Kits and the FLIPR^TETRA System.  
• LiveWare reagents have been successfully used on the FLIPR System, in conjunction with non-G_q-coupled members of chemokine, serotonin, glutamate, dopamine, opoid, vasopressin and a- and b-adrenoceptor receptor families. 
• Receptor configuration options are increased as LiveWare products are available in the following configurations (plasmids are available as hygromycin and neomycin (G418)-resistant; cells are hygromycin-resistant*:
• Host cells: CHO cells stably transfected with one of the G-protein chimera genes or the G_α16 gene.
• Plasmids: Replicating G-protein expression vectors. 

*Each plasmid order contains one vial of hygromycin-resistance plasmid and one vial of neomycin (G418)-resistance plasmid.

Specifications

Information about LiveWare cells and plasmids is listed in the chart below. Click on the Product # to view a product information sheets.  Vector maps and sequences are available by request from technical support or your sales representative.

LiveWare Product Specifications

For information about sequences or vector maps,  please contact Technical Support or your Sales Representative.

Applications

Figure 1. Chimeric Gqi5 couples CCR5 chemokine receptor to calcium response

Figure 2. Gα16 couples b-adrenergic receptor to calcium response

Figure 3. Chimeric and promiscuous G-proteins couple k-opioid receptor to calcium response

Ordering Information

Part numbers for ordering can be found in the Product Specifications Table above.