The ACSL5 Knockout SK-HEP-1 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human SK-HEP-1 hepatocellular carcinoma cell line. It introduces a loss-of-function model for ACSL5 (acyl-CoA synthetase long-chain family member 5), a critical enzyme in hepatic lipid metabolism. The polyclonal pool retains heterogeneous knockout profiles, avoiding clonal selection bias and representing population-level gene inactivation.
SK-HEP-1 is a well-established human hepatocellular carcinoma line originally isolated from the ascites of a liver adenocarcinoma patient. These epithelial cells maintain key hepatic metabolic features and are widely used in hepatocarcinogenesis, metabolic reprogramming, and drug metabolism studies. Their metastatic origin enhances relevance for invasive liver cancer research. The ACSL5 knockout in this context enables direct analysis of fatty acid activation’s role in tumor cell biology.
ACSL5 catalyzes the ATP-dependent thioesterification of long-chain fatty acids, such as palmitate, with coenzyme A to form fatty acyl-CoA esters. These activated intermediates are channeled either into mitochondrial ??-oxidation via CPT1A or into complex lipid synthesis, including triglyceride and phospholipid production. ACSL5 expression is controlled by transcription factors PPAR??, SREBP-1c, and LXR, and is responsive to insulin signaling. It interacts with fatty acid-binding protein 1 (FABP1) and associates with lipid droplet surfaces, where it generates acyl-CoA for droplet expansion. Disruption of ACSL5 therefore profoundly alters fatty acid partitioning between oxidation and storage, impacting overall energy homeostasis.
In hepatocellular carcinoma, enhanced fatty acid utilization supports proliferation. ACSL5 knockout in SK-HEP-1 cells disrupts this metabolic adaptation, impairing conversion of exogenous fatty acids to energy and lipids. This model is valuable for studying lipotoxicity??where unesterified fatty acids trigger ER stress and apoptosis??and for exploring links to non-alcoholic fatty liver disease and insulin resistance. The polyclonal population reveals compensatory mechanisms, better mimicking tumor heterogeneity and providing insights into therapeutic vulnerabilities.
Typical applications include quantitative fatty acid oxidation assays using radiolabeled or fluorescent palmitate, comprehensive lipidomics profiling via LC?CMS to quantify changes in acyl-CoA and complex lipid species, and ATP bioluminescence assays to gauge metabolic output. Apoptosis induction can be monitored by Annexin V staining, while RT-qPCR and western blotting confirm ACSL5 disruption and evaluate compensatory gene expression. This knockout product serves as an essential tool for dissecting the intersection of lipid metabolism and oncogenic signaling, identifying lipid-related drug targets, and testing candidate therapeutics that modulate fatty acid handling in liver cancer. For further technical information, please contact Ascent Research.
