The ACADVL Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, with targeted disruption of the ACADVL gene. Supplied as a heterogeneous pool, this product avoids single-cell cloning artifacts and enables robust population-level loss-of-function studies, validated for ACADVL inactivation at the pooled level.
The HT29 host cell line originates from a grade I human colon adenocarcinoma (1964) and is a widely used epithelial model for colorectal cancer research. HT29 cells form polarized monolayers, express intestinal epithelial markers, and exhibit stable growth characteristics, making them an ideal platform to study metabolic regulation, lipid metabolism, and mitochondrial function in a transformed colorectal context.
ACADVL encodes very long-chain acyl-CoA dehydrogenase (VLCAD), a mitochondrial flavoenzyme that catalyzes the initial dehydrogenation step in the beta-oxidation of long-chain fatty acids (C14?CC20). The enzyme uses FAD as its cofactor and transfers electrons to electron transfer flavoprotein (ETF), thereby coupling fatty acid degradation to the respiratory chain. ACADVL expression is transcriptionally regulated by PPAR?? and its co-activator PGC-1??, and the enzymatic reaction functions downstream of the carnitine shuttle components CPT1A and CPT2. ACADVL activity precedes that of downstream beta-oxidation enzymes such as medium-chain acyl-CoA dehydrogenase (ACADM) and the mitochondrial trifunctional protein subunits HADHA and HADHB. Disruption of ACADVL blocks the oxidation of long-chain fatty acids, leading to accumulation of long-chain acylcarnitines and free fatty acids, impaired ketogenesis, and severe energy deficiency, particularly under fasting or metabolic stress conditions, due to reduced electron flux through ETF and diminished acyl-CoA intermediate supply.
Within the HT29 colorectal cancer model, ACADVL knockout recapitulates features of VLCAD deficiency, creating a reliance on alternative substrates and exposing metabolic vulnerabilities. Colorectal tumors frequently rewire lipid metabolism; loss of ACADVL forces compensatory shifts that can be probed for synthetic lethal interactions. Given that beta-oxidation-derived acetyl-CoA and NADH support anabolic pathways and redox balance, ACADVL deficiency in HT29 cells reveals how colorectal cancers adapt to mitochondrial lipid oxidation defects. This polyclonal model thus allows investigation of how fatty acid oxidation supports cancer cell proliferation, survival under nutrient limitation, and maintenance of redox homeostasis, while also providing a physiologically relevant system to study epithelial consequences of acylcarnitine accumulation.
Typical applications include acylcarnitine profiling by LC-MS to quantify metabolic intermediates, fatty acid oxidation rate assays using labeled palmitate, and mitochondrial stress tests via Seahorse analysis to measure oxygen consumption rates. The model is well-suited for cell viability assays under glucose deprivation or metabolic stress, RT-qPCR and western blotting for ACADVL, PPAR?? targets, and apoptosis markers, and proliferation/apoptosis assessments. Additionally, it supports research into metabolic reprogramming in response to pharmacological PPAR?? agonists or mitochondrial uncouplers, and the interplay between lipid catabolism and oncogenic signaling. For further product information or technical inquiries, please contact Ascent Research.
