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GPCRs: G Protein-Coupled Receptors
G Protein-Coupled Receptors (GPCRs) are a large and diverse group of membrane proteins that play a critical role in cellular communication and signal transduction. Involved in a wide range of physiological processes, GPCRs are among the most studied drug targets, with more than 30% of FDA-approved drugs targeting them.
Types of GPCRs and Their Functions:
Gi-GPCRs: Regulate adenylyl cyclase activity by inhibiting cAMP production, leading to inhibitory cellular effects.
Gs-GPCRs: Regulate adenylyl cyclase activity by stimulating cAMP production, leading to stimulatory cellular effects.
Gq-GPCRs: Activate PLC, leading to calcium release and PKC activation.
G12/13-GPCRs: Regulate the cytoskeleton and cell motility through Rho GTPases.
ACTOne GPCR Assay
ACTOne™ is a high-throughput GPCR screening technology that measures intracellular changes of the secondary messenger cyclic AMP (cAMP) in living cells, in real-time. The assay uses a proprietary modified cyclic nucleotide-gated (CNG) channel, co-localized with adenylate cyclase at the plasma membrane, as a biosensor of cAMP activity. When cAMP levels increase, the CNG channel opens, resulting in ion flux and cell membrane depolarization.
Key Features:
Real-Time Measurement: Quantifies intracellular cAMP changes using fluorescent ion indicators or membrane potential (MP) dyes.
High Sensitivity: Detects both strong and weak GPCR activities with minimal artifacts.
Versatile Applications: Suitable for detecting endogenous GPCR responses and activities not reported in the literature.
Advantages of ACTOne GPCR Assay: Directly measures GPCR activation. Provides both kinetic and endpoint readouts. Greater content and statistically relevant data.
AAV ACTOne GPCR Assay Kits
GPCR stable cell lines are widely used for drug screening due to their reliability and consistency in expressing GPCRs. These cell lines are engineered to stably express a specific GPCR, making them ideal for high-throughput screening of potential drug candidates. The use of GPCR stable cell lines allows researchers to efficiently evaluate the pharmacological properties of compounds, including their potency, efficacy, and selectivity. However, the stable cell lines has limited receptor diversity. Developing and validating a stable GPCR cell line can be time-consuming and costly, particularly for difficult-to-express receptors or those requiring specific post-translational modifications. Moreover, stable cell lines typically express only one type of GPCR, whereas native tissues often co-express multiple GPCRs and other receptors that can interact and influence each other.
In the AAV ACTOne GPCR Assay Kit, we use AAV vector to express both CNG and GPCR. It offers significant advantages over stable cell lines, including faster setup, greater physiological relevance, reduced risk of genetic drift, simplified workflows, flexibility in co-expression, dynamic control of expression, and cost and time efficiency. These benefits make AAV ACTOne GPCR Assay Kit a powerful alternative for GPCR drug screening and research.
Components in AAV ACTOne GPCR Assay Kits
Each kit includes two main components:
AAV-CNG/GPCR enables transient expression of both CNG and GPCRs. AAV-CNG/GPCR is a mixture of AAV-CNG and AAV-GPCR. Both viruses use CMV as promoter, which is known for driving strong and rapid expression of transgenes in a wide variety of cell types. The ratio of two viruses are optimized in each kit.
ACTOne MP Dye allows for real-time, sensitive detection of GPCR functionality.
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Advantages of AAV ACTOne GPCR Assay Kits
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