The ADCY3 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the ADCY3 gene has been disrupted to create a loss-of-function model for cAMP signaling studies. This product provides a mixed population of HT29 cells carrying targeted gene disruption, allowing researchers to investigate the role of adenylyl cyclase 3 in colorectal cancer biology without the need for single-cell cloning.
The HT29 host cell line is derived from a human colorectal adenocarcinoma and exhibits adherent epithelial morphology with a hypertriploid karyotype. HT29 cells are widely utilized as a model for intestinal epithelial function and colorectal cancer, as they retain key characteristics of the colonic epithelium and respond to various hormonal and metabolic stimuli. Their ease of culture and well-characterized signaling pathways make them a robust platform for interrogating GPCR-mediated signal transduction.
ADCY3 encodes a membrane-associated adenylyl cyclase that catalyzes the conversion of ATP to cyclic AMP (cAMP) upon activation by Gs-coupled GPCRs such as GLP1R, MC4R, and ADRB2. The Gs alpha subunit, along with cofactors like calcium/calmodulin and forskolin, stimulates ADCY3 activity. The resulting cAMP activates downstream effectors PKA and Epac, which phosphorylate transcription factors like CREB and modulate MAPK signaling. ADCY3 also interacts with calmodulin, AKAP5, G?¦?, and Ric8A, and regulates ion channels such as HCN and CNG, thereby integrating multiple signaling inputs to control cellular responses.
In HT29 colorectal cancer cells, ADCY3 knockout disrupts the canonical cAMP-PKA-CREB axis, impairing receptor-stimulated cAMP accumulation and altering the phosphorylation of CREB at Ser133. This loss of function also impacts Epac-dependent pathways, including Rap1-mediated MAPK activation, which can affect cell proliferation, migration, and metabolic reprogramming. Given the association of ADCY3 with obesity, type 2 diabetes, and colorectal cancer, this knockout model is a valuable tool for dissecting the molecular mechanisms linking cAMP signaling to tumorigenic processes in the gut.
This polyclonal knockout product is ideally suited for a range of experimental applications, including cAMP signaling investigation in colorectal cancer, GPCR pharmacological profiling, metabolic disorder modeling, and drug screening for obesity and diabetes. Researchers can employ assays such as cAMP accumulation measurements (ELISA or FRET-based), Western blotting for phospho-CREB (Ser133), RT-qPCR analysis of CREB target genes like NR4A1 and FOS, MTT proliferation assays, transwell migration/invasion assays, phospho-kinase antibody arrays, and flow cytometry for cell cycle analysis. For further information or to acquire these cells, please contact Ascent Research.
