The ADK Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma cell line, designed for functional investigation of adenosine kinase (ADK). This product features a heterogeneous pool of cells carrying targeted disruptions in the ADK gene, resulting in a loss-of-function model without clonal selection. The polyclonal format preserves genetic diversity while ensuring knockout of ADK, enabling robust modeling of adenosine signaling perturbations in cancer research. Researchers can employ this tool to dissect the role of ADK in purine metabolism and downstream pathways, with applications spanning drug discovery, signal transduction, and epigenetic regulation.
HCT 116 is a widely utilized colorectal carcinoma epithelial cell line characterized by a KRAS G13D mutation, microsatellite stability (MSS), and a near-diploid karyotype, making it a representative model for colon cancer biology. These cells exhibit adherent growth and are extensively employed to study oncogenic signaling, tumor progression, and therapeutic responses. The HCT 116 background provides a clinically relevant context for examining ADK function, as adenosine metabolism is increasingly implicated in colorectal cancer progression and immune evasion. The combination of the defined genomic profile of HCT 116 with ADK knockout creates a powerful system to explore how aberrant adenosine homeostasis influences malignant phenotypes.
ADK functions as the primary enzyme for adenosine clearance by catalyzing the phosphorylation of adenosine to AMP, thereby tightly regulating intra- and extracellular adenosine concentrations. This activity directly controls adenosine receptor signaling through ADORA1, ADORA2A, ADORA2B, and ADORA3, and intersects with the methionine cycle via modulation of S-adenosylmethionine (SAMe) levels and DNA methylation. Knockout of ADK leads to accumulation of extracellular adenosine, which activates adenosine receptors and engages downstream effectors including AMPK and cAMP signaling. Key upstream regulators such as inflammatory cytokines (TNF, IL1B) and hypoxia-inducible factor 1A (HIF1A) modulate ADK expression, while interacting factors like S-adenosylhomocysteine hydrolase (AHCY) and importin ??5 (KPNA1) coordinate its subcellular localization and metabolic coupling. This network positions ADK at the nexus of purine salvage, methionine metabolism, and receptor-mediated cellular responses.
In the HCT 116 colon cancer model, ADK knockout is anticipated to elevate extracellular adenosine levels, leading to sustained adenosine receptor activation and subsequent alterations in cell proliferation, migration, and epigenetic states. The KRAS mutant background may synergize with adenosine-driven signaling to modulate tumor aggressiveness and the inflammatory microenvironment. By disrupting adenosine homeostasis, these polyclonal knockout cells provide a platform to investigate how adenosine receptor subtypes (A1, A2A, A2B, A3) contribute to colorectal carcinoma pathophysiology and to identify potential vulnerabilities for therapeutic intervention targeting the adenosine axis. The model is particularly suited for studies linking metabolic reprogramming to cancer cell plasticity.
This product supports a diverse array of experimental approaches, including quantitative assessment of ADK expression via Western blotting and RT-qPCR, measurement of adenosine and AMP levels by HPLC, and functional assays for adenosine receptor activity using cAMP detection and AMPK phosphorylation analysis. Cell proliferation (MTS/XTT), migration, and invasion assays enable phenotypic characterization, while RNA-seq and ATP measurement facilitate transcriptomic and metabolic profiling. Applications encompass validation of adenosine-modulating drug candidates, screening of receptor agonists/antagonists, and exploration of methionine cycle-dependent epigenetic regulation in colorectal cancer. For additional technical specifications or ordering inquiries, please contact Ascent Research.
