The ADCY5 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered from the HCT 116 human colorectal carcinoma cell line, designed to disrupt the ADCY5 gene and enable loss-of-function analyses. This product consists of a heterogeneous pool of cells carrying targeted gene disruption at the ADCY5 locus, providing a versatile tool for studying adenylyl cyclase 5 biology without the need for clonal isolation. As a polyclonal population, it retains the diversity of editing outcomes, which can be advantageous for assessing overall pathway effects in a physiologically relevant cancer model.
HCT 116 is a well-characterized KRAS-mutant colorectal carcinoma cell line widely used as an in vitro model for colorectal cancer research. These cells exhibit an epithelial morphology and harbor an activating KRAS mutation that drives constitutive MAPK signaling, contributing to uncontrolled proliferation, survival, and metabolic reprogramming. The KRAS-mutant background makes HCT 116 particularly valuable for investigating cross-talk between RAS-driven and cAMP-dependent pathways, as well as for evaluating therapeutic vulnerabilities.
ADCY5 encodes type 5 adenylyl cyclase, a membrane-bound enzyme that converts ATP to cAMP upon Gs protein-coupled receptor activation. Upstream receptors such as beta-adrenergic, prostaglandin E2, and adenosine receptors stimulate ADCY5 via Gs/olf proteins. Downstream, cAMP activates effectors including PKA, EPAC, and CREB. ADCY5 activity is further modulated by interactions with G?¦?, calmodulin, and protein kinase C. In colorectal cancer cells, cAMP signaling regulates phosphorylation events that influence proliferation, migration, and apoptosis.
In the KRAS-mutant colorectal cancer context, ADCY5-mediated cAMP generation can modulate tumorigenic processes, including cell cycle progression, survival, and motility. By using these polyclonal knockout cells, researchers can interrogate how loss of ADCY5 alters cAMP dynamics and downstream PKA/CREB transcriptional programs, potentially unmasking synthetic vulnerabilities or resistance mechanisms to targeted therapies. The HCT 116 model is particularly suited for such studies due to its extensive characterization and the availability of complementary data on RAS pathway interactions.
This ADCY5 knockout product supports a wide range of experimental applications, such as investigating GPCR-cAMP-PKA signaling in colorectal cancer, elucidating the role of adenylyl cyclase 5 in tumorigenesis, and screening for small-molecule modulators of the cAMP pathway. Typical assays include western blotting for phosphorylated PKA substrates, cAMP enzyme-linked immunosorbent assays (ELISA), reverse transcription quantitative PCR (RT-qPCR) for CREB target genes, cell proliferation and colony formation assays, migration/invasion tests, and drug sensitivity profiling. For additional information, please contact Ascent Research.
