AAV ACTOne Gi-GPCR Assay Kit-S1PR2

Sphingosine-1-Phosphate Receptor 2 (S1PR2) is a G protein-coupled receptor (GPCR) that binds to the bioactive lipid sphingosine-1-phosphate (S1P). S1PR2 is one of five receptors (S1PR1-5) in the S1P receptor family, each of which mediates various physiological functions through interactions with S1P. S1PR2 is primarily coupled with multiple G proteins, including G12/13, Gq, and Gi, allowing it to activate diverse downstream signaling pathways. When S1PR2 is activated by S1P, it can lead to the activation of several signaling cascades, including the Rho/Rho kinase pathway (via G12/13), the phospholipase C (PLC) pathway (via Gq), and inhibition of adenylate cyclase (via Gi). These pathways collectively influence cell morphology, migration, proliferation, and survival. One of the key roles of S1PR2 is regulating cytoskeletal organization and cell migration, often resulting in the inhibition of cell movement, which contrasts with the effects of S1PR1. S1PR2 is expressed in various tissues, including the cardiovascular system, central nervous system, and immune system. In the cardiovascular system, S1PR2 is involved in vascular development, endothelial barrier function, and smooth muscle cell contraction. In the central nervous system, it has been implicated in regulating neuronal signaling and brain development. S1PR2 also plays roles in immune cell trafficking and the inflammatory response. Due to its involvement in diverse biological processes, S1PR2 has been implicated in several pathological conditions, including cancer, fibrosis, and immune disorders. For instance, in cancer, S1PR2 can influence tumor progression and metastasis by regulating cell migration and survival. In the context of fibrosis, S1PR2 signaling contributes to the excessive deposition of extracellular matrix proteins, leading to tissue scarring. Research into S1PR2 continues to explore its therapeutic potential, particularly in diseases where abnormal cell migration, proliferation, and immune responses are prominent.
This kit uses AAV vectors with a CMV promoter to co-express the S1PR2 and cyclic nucleotide-gated (CNG) channel, allowing researchers to conduct high-throughput screening and functional analysis of potential S1PR2-targeting compounds. The kit provides a sensitive and reliable method for evaluating the pharmacological properties of S1PR2 drugs, such as agonists and antagonists, in a live-cell environment.

Please contact us at for specific academic pricing.

Background

ACTOne™ is the only high-throughput GPCR screening technology that can directly measure the intracellular changes of the secondary messenger cyclic AMP (cAMP) in living cells, in real-time. It uses a proprietary modified cyclic nucleotide-gated (CNG) channel, which is co-localized with adenylate cyclase at the plasma membrane, as a biosensor of cAMP activity. The CNG channel opens when the cAMP level near the plasma membrane increases, resulting in ion flux and cell membrane depolarization. The influx of cations through the CNG channel can be quantified using fluorescent ion indicators or membrane potential (MP) dyes. It provides information on real time intracellular cAMP changes and is highly sensitive. By combining kinetic and endpoint readouts, we are able to capture and analyze transient responses from endogenous GPCRs and weak responses caused by weak Gs or Gi coupled GPCR activities. Using ACTOne, we are able to detect the subcellular cAMP concentration changes directly caused by GPCR activation. Real-time kinetic readouts minimize artifacts, and provide greater content and more statistically relevant data. The intensity of signal increase caused by GPCR activation is directly related to the receptor number on cell surface. Using ACTOne assay, we were able to detect activities of some endogenous Gs coupled receptors in HEK293 cells that have not been reported in literature. In addition, we have also detected weak Gs coupled activity of a GPCR that was widely considered to be only linked to Gq coupled pathway. The ACTOne assay also provides a useful tool for GPCR de-orphanization.