ADRB2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the human ADRB2 gene. This product provides a heterogeneous pool of edited HEK293T cells with targeted disruption of the beta-2 adrenergic receptor locus, enabling investigation of ADRB2-dependent signaling without clonal artifacts. The polyclonal format captures a range of genetic edits, offering a robust model for studying overall gene ablation effects in a widely used human cell background.
The HEK293T cell line is a well-characterized human embryonic kidney epithelial line immortalized by expression of SV40 large T antigen, which supports high-level episomal plasmid replication and efficient transfection. Widely adopted for protein expression, viral packaging, and receptor pharmacology, HEK293T provides a versatile platform for dissecting GPCR signaling pathways. Its epithelial origin and amenability to a variety of genetic manipulations make it particularly suited for studying receptor function, trafficking, and downstream cascades in a human cellular context.
The ADRB2 gene encodes the beta-2 adrenergic receptor, a classical Gs-coupled GPCR activated by endogenous catecholamines such as epinephrine and norepinephrine. Agonist binding promotes receptor coupling to the Gs alpha subunit, which stimulates adenylyl cyclase to generate cAMP. Elevated cAMP activates protein kinase A (PKA), which phosphorylates numerous substrates including CREB, phospholamban, and glycogen phosphorylase, driving responses like bronchodilation, glycogenolysis, and lipolysis. Additionally, GPCR kinase (GRK2)-mediated phosphorylation recruits ??-arrestin 1/2, leading to receptor desensitization, internalization, and G protein-independent activation of the ERK1/2 MAPK cascade. The receptor also interacts with A-kinase anchoring proteins (AKAPs) and phosphodiesterase PDE4D to fine-tune localized cAMP signaling.
In HEK293T cells, endogenous ADRB2 expression enables reconstitution of the full signaling module from receptor to transcriptional output. By disrupting ADRB2, this knockout model eliminates both G protein-dependent and ??-arrestin-mediated branches, allowing rigorous interrogation of pathway selectivity. The absence of the receptor eliminates agonist-induced cAMP accumulation, PKA substrate phosphorylation, and CREB-dependent transcription, while preserving responsiveness of other GPCRs. This controlled genetic background is invaluable for mapping ADRB2-specific contributions to cAMP/PKA dynamics, testing biased agonism, and validating target engagement of beta-2 ligands in an isogenic system.
Typical research applications include functional characterization of beta-2 adrenergic receptor signaling pathways, high-throughput screening of beta-2 agonists and antagonists, mechanistic studies of GPCR desensitization and internalization, and dissection of cAMP/PKA-driven transcriptional programs. The knockout cells can be employed in cAMP accumulation assays, radioligand binding studies, phospho-PKA substrate western blotting, CREB target gene qPCR, and ??-arrestin recruitment bioluminescence resonance energy transfer (BRET) experiments. They also serve as a negative control for validating antibody specificity or siRNA effects. This product is suitable for asthma, COPD, heart failure, and metabolic disease research. For further information, please contact Ascent Research.
