The ADCY6 Knockout A-549 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal population derived from the A-549 human lung adenocarcinoma cell line, in which the ADCY6 gene has been disrupted to create a loss-of-function model. This heterogeneous knockout pool avoids the limitations of single?cell clones and enables consistent interrogation of ADCY6-mediated pathways in a physiologically relevant epithelial context, providing a robust platform for cAMP signaling research.
The parental A-549 line originates from human lung adenocarcinoma and serves as a model of alveolar type II epithelium. These cells carry a KRAS mutation and wild?type p53, reflecting common genetic alterations in non-small-cell lung cancer. A-549 cells retain key epithelial characteristics, including ion transport functionality and responsiveness to GPCR agonists, making them a relevant host for ADCY6 knockout studies.
ADCY6 encodes adenylyl cyclase 6, which catalyzes cAMP production downstream of Gs-coupled receptors such as ADRB2 and PTGER2/4 via GNAS. The generated cAMP activates PKA and EPAC1/2: PKA phosphorylates CREB and regulates CFTR, while EPAC activates RAP1. ADCY6 interacts with GNAS, GNAI2, AKAP5, calmodulin, and ???arrestin?2, which regulate its localization and activity. Key signaling routes include ADRB2??GNAS??ADCY6??cAMP??PKA??CREB and PTGER2??GNAS??ADCY6??cAMP??EPAC??RAP1. Knockout of ADCY6 therefore impairs cAMP-dependent PKA/CREB and EPAC pathways, altering downstream gene transcription, ion transport, and cellular responses.
In A?549 lung adenocarcinoma cells, ADCY6 knockout disrupts cAMP?regulated processes critical to epithelial function, including CFTR?mediated ion transport, CREB?driven transcription, and anti?apoptotic signaling. The KRAS?mutant background may reveal crosstalk between cAMP and oncogenic pathways, influencing proliferation, survival, and metastasis. Because A?549 cells respond to ???adrenergic agonists and prostaglandins, this model is valuable for studying GPCR?cAMP axis dysfunction in asthma, lung injury, and drug validation aimed at restoring cAMP homeostasis.
Typical applications include respiratory disease modeling, cAMP signaling network dissection, drug target validation for asthma and cardiac arrhythmias, host?Cpathogen interaction studies, and lung cancer proliferation/apoptosis assays. Validation methods encompass western blotting and RT?qPCR for ADCY6, cAMP accumulation and PKA activity assays, CREB phosphorylation and CFTR immunofluorescence, flow cytometry for apoptosis, and MTT/BrdU proliferation assays; drug sensitivity and migration assays further expand utility. For product specifications or technical inquiries, please contact Ascent Research.
