ADK Knouckout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human epithelial colorectal adenocarcinoma cell line HT29. This product, targeting the adenosine kinase (ADK) gene, provides a loss-of-function model through Cas9-mediated gene disruption, enabling the study of ADK-dependent purine metabolism and adenosine signaling. The polyclonal nature of the knockout population ensures robust representation of edited alleles while maintaining the heterogeneous background typical of HT29 cultures.
The HT29 cell line, originally isolated from a 44-year-old female Caucasian with colorectal adenocarcinoma, is an extensively characterized model of the intestinal epithelium. HT29 cells are capable of enterocytic differentiation under appropriate conditions, making them valuable for investigations of intestinal barrier function, colorectal cancer biology, and tumor microenvironment interactions. Their well-documented signaling networks and growth properties render them a reliable host for functional gene knockout studies.
ADK encodes adenosine kinase, which catalyzes the phosphorylation of adenosine to AMP, thereby serving as a principal regulator of intracellular and extracellular adenosine concentrations. This enzyme is upregulated by hypoxia, TNF-alpha, IL-1beta, cAMP, and HIF-1alpha, and its activity directly modulates adenosine receptor (A1, A2A, A2B, A3) signaling, AMPK and mTOR pathways, and cAMP-dependent effectors. In the presence of Mg2+, ADK interacts with adenosine and ATP to control the balance of purine salvage, influencing downstream cascades including adenylate cyclase?CcAMP?CPKA and AMPK-mediated metabolic responses.
In the HT29 model, disruption of ADK results in adenosine accumulation, which potentiates adenosine receptor activation and perturbs cAMP homeostasis. This alteration can affect cell proliferation, migration, and immune regulatory functions, as adenosine signaling plays critical roles in tumor growth and immune evasion within the colorectal cancer microenvironment. The ADK knockout thus offers a physiologically relevant system to dissect how adenosine-driven pathways intersect with colorectal adenocarcinoma progression.
Researchers can employ these cells in assays such as Western blotting, RT-qPCR, LC-MS/MS adenosine quantification, cAMP measurement, cell proliferation and migration assays, phospho-signaling analysis, and flow cytometry-based apoptosis detection. Applications span adenosine signaling studies, cancer metabolism research, immunomodulation assays, drug target validation for epilepsy and inflammatory diseases, and colorectal cancer progression modeling. For further details on experimental utilization and technical specifications, please contact Ascent Research.
