The ADCY6 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, targeting the ADCY6 gene. This loss-of-function model enables the study of adenylyl cyclase 6-dependent signaling in a colorectal epithelial context. The polyclonal pool contains cells with diverse CRISPR-induced mutations, making it suitable for bulk biochemical assays, pooled functional screens, and pathway analysis where clonal uniformity is not essential. This product provides a robust tool for interrogating cAMP-mediated processes without endogenous ADCY6 activity.
The HT29 cell line is a well-established model of human colorectal adenocarcinoma, originally isolated from a primary tumor in 1964. These epithelial cells recapitulate features of the intestinal epithelium and are widely used in studies of intestinal cell differentiation, cancer biology, and drug sensitivity. Their characterized signaling networks and ease of genetic manipulation make them an ideal host for creating knockout models. In this polyclonal ADCY6-knockout background, HT29 cells maintain their epithelial phenotype while lacking functional cyclase activity, allowing focused investigation of ADCY6??s role in colorectal cancer.
ADCY6 encodes adenylyl cyclase 6, a membrane-bound enzyme that converts ATP to the second messenger cAMP upon activation by G??s-coupled receptors such as the ??-adrenergic receptor and prostaglandin E2 receptor. Its activity is further modulated by calmodulin, protein kinase C (PKC), and calcium ions. cAMP produced by ADCY6 activates downstream effectors including PKA, which phosphorylates CREB and other substrates, and EPAC, which triggers Rap1 and MAPK/ERK cascades. ADCY6 interacts with G??i subunits, ??-arrestin, and AKAPs, integrating GPCR signals to regulate proliferation, differentiation, and apoptosis. Disruption of ADCY6 impairs this signaling node, providing a system to dissect cAMP-dependent pathways in colorectal cancer.
In HT29 colorectal adenocarcinoma cells, ADCY6 knockout disrupts GPCR-mediated cAMP production, compromising the PKA-CREB and EPAC-Rap1 signaling axes. This model is particularly valuable for elucidating the contributions of cAMP to colorectal tumorigenesis, where dysregulated adenylyl cyclase activity influences cell growth, survival, and migration. By comparing knockout and wild-type cells, researchers can isolate isoform-specific functions and explore compensatory mechanisms within the adenylyl cyclase family. The polyclonal population also mirrors the heterogeneity of tumor cells, enhancing physiological relevance.
This ADCY6 knockout model supports diverse research applications, including mechanistic studies of cAMP signaling in colorectal cancer, GPCR pathway investigation in intestinal biology, and screening of adenylyl cyclase modulators. Recommended assays include cAMP quantification (ELISA, FRET-based sensors), Western blotting for phospho-CREB, cell proliferation (MTT, BrdU) and apoptosis (Annexin V) measurements, RT-qPCR for CREB target genes, and migration/invasion assays. Phospho-signaling arrays can reveal pathway rewiring upon ADCY6 loss. For additional information or to acquire this product, contact Ascent Research.
