The ADH1B Knockout HCT 116 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which the ADH1B gene has been disrupted within the human HCT 116 colorectal carcinoma cell line. This product offers a heterogeneous pool of gene-edited cells for functional studies of alcohol dehydrogenase 1B, enabling researchers to examine the loss-of-function consequences in a genetically defined cancer model. The polyclonal format provides a robust experimental system without the limitations of single-cell clonal isolates, making it suitable for a wide range of biochemical and cell-based assays.
HCT 116 is a well-characterized, near-diploid human epithelial cell line derived from a colorectal adenocarcinoma. Its genomic landscape includes an activating KRAS(G13D) mutation, a stabilizing CTNNB1 (??-catenin) mutation, and microsatellite instability-high (MSI-H) status due to defective DNA mismatch repair. These alterations drive constitutive MAPK and Wnt signaling, promoting uncontrolled proliferation. The line is extensively used in colorectal cancer research, intestinal epithelial biology, and drug discovery, offering a relevant host context for probing metabolic gene function in an oncogenic background.
ADH1B encodes an enzyme that catalyzes the oxidation of ethanol to acetaldehyde, a reactive and potentially carcinogenic intermediate. This reaction proceeds with the reduction of NAD? to NADH, linking ethanol metabolism to cellular redox balance and energy pathways such as glycolysis and gluconeogenesis. ADH1B also participates in retinol metabolism by oxidizing retinol to retinal. The enzyme is transcriptionally regulated by hepatocyte nuclear factor 4 alpha (HNF4A) and CCAAT/enhancer-binding protein alpha (CEBPA), and its activity is induced by substrates like ethanol and retinoic acid. Downstream, acetaldehyde can form DNA adducts and generate reactive oxygen species (ROS), while NADH accumulation shifts the NAD?/NADH ratio. ADH1B functionally interacts with aldehyde dehydrogenase 2 (ALDH2), which detoxifies acetaldehyde, and with cytochrome P450 2E1 (CYP2E1) and alcohol dehydrogenase 1A (ADH1A), which provide parallel ethanol-oxidizing routes. Catalase also participates in ethanol degradation. Disruption of ADH1B therefore uncouples this metabolic network.
In the HCT 116 model, ADH1B knockout is particularly informative given the host cell??s oncogenic drivers. The KRAS(G13D) mutation sustains proliferative signaling, while ??-catenin stabilization promotes transcriptional programs that may intersect with acetaldehyde-induced stress responses. The MSI-H phenotype predisposes cells to DNA damage sensitivity, making the knockout a powerful tool to study how loss of ethanol metabolism influences genomic instability, redox homeostasis, and apoptotic thresholds. This model can reveal context-dependent roles of ADH1B in colorectal carcinoma, potentially linking alcohol metabolism to tumor cell survival and DNA repair.
This polyclonal knockout cell product is ideally suited for a range of functional assays. Researchers can quantify ethanol conversion rates, measure acetaldehyde accumulation, assess ROS levels using fluorescent probes, and monitor NADH/NAD? ratios via enzymatic cycling. DNA damage can be evaluated by comet assay and ??-H2AX immunostaining, while cell fate changes are analyzed through apoptosis and colony formation assays. These applications position the ADH1B Knockout HCT 116 Polyclonal Cells as a versatile platform for investigating ethanol-induced carcinogenesis, oxidative stress, NAD? metabolism, and drug metabolism in a colorectal cancer context. For more information, please contact Ascent Research.
