The ADCY9 Knouckout HT29 Polyclonal Cells product provides a mixed population of HT29 colorectal adenocarcinoma cells with CRISPR/Cas9-mediated disruption of ADCY9. This polyclonal knockout model avoids clonal selection artifacts, offering a heterogeneous loss-of-function system for studying adenylyl cyclase 9-dependent signaling in a disease-relevant cellular background.
HT29 cells are derived from a human colorectal adenocarcinoma and carry well-defined oncogenic mutations, including mutant APC, BRAF V600E, and TP53. These alterations drive aberrant Wnt, MAPK, and p53 pathways, rendering HT29 a widely used model for colorectal cancer research. The epithelial morphology and moderate differentiation of HT29 enable assessment of how ADCY9 inactivation impacts proliferation, differentiation, and apoptosis in a genetically characterized context.
ADCY9 encodes a membrane-bound adenylyl cyclase that catalyzes conversion of ATP to cAMP upon stimulation by Gs-coupled GPCRs, exemplified by the beta2-adrenergic receptor. Its activity is regulated by the Gs alpha subunit, calcium/calmodulin, and protein kinase C. The generated cAMP activates PKA and EPAC (RAPGEF3/4), leading to phosphorylation of CREB and other effectors. ADCY9 signaling is organized by interacting partners such as AKAPs, beta-arrestins, and calmodulin, and is terminated by phosphodiesterases. In HT29 cells, this cascade influences cell cycle progression, survival, and metabolic responses, intersecting with oncogenic pathways.
In HT29 colorectal adenocarcinoma, ADCY9-dependent cAMP signaling modulates cellular responses to external stimuli and pharmacological agents. Since ADCY9 expression is altered in colorectal cancer, the knockout model allows dissection of its role in tumor biology. The polyclonal population facilitates examination of ADCY9-specific effects on proliferation, differentiation, and drug sensitivity, unaffected by clonal compensation. This system is especially useful given the cell line’s reliance on BRAF V600E and Wnt-driven signaling.
The ADCY9 knockout polyclonal cells support numerous applications, including Western blot and RT-qPCR for knockout verification, cAMP accumulation assays, PKA substrate and CREB phosphorylation analyses, and immunofluorescence. The model is well-suited for GPCR agonist/antagonist profiling, drug sensitivity screens (e.g., dalcetrapib), and studies of cAMP-mediated differentiation or apoptosis. Proliferation and viability assays further enable interrogation of ADCY9 function in colorectal cancer. For technical inquiries, contact Ascent Research.
