The HACL1 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human hepatocellular carcinoma line, with disruption of the HACL1 gene. This loss-of-function model enables investigation of peroxisomal alpha-oxidation without assumptions of clonality or specific editing patterns. The polyclonal format provides a heterogeneous pool of edited alleles, reflecting genetic variability relevant to disease modeling while ensuring effective target-gene disruption at the population level.
SK-HEP-1 is a well-established hepatocellular carcinoma cell line, originally isolated from the ascitic fluid of a liver adenocarcinoma patient. As a cancerous liver epithelial line, SK-HEP-1 maintains hepatic metabolic functions, including peroxisomal activity, and is extensively used to study lipid metabolism, hepatocarcinogenesis, and metabolism-driven cancer phenotypes. Its tumorigenic properties make it a relevant host for exploring the role of HACL1 in the context of liver cancer.
HACL1 encodes 2-hydroxyacyl-CoA lyase 1, a peroxisomal enzyme that catalyzes TPP-dependent cleavage of 2-hydroxyphytanoyl-CoA to formyl-CoA and pristanal during alpha-oxidation. This step is essential for phytanic acid breakdown. HACL1 expression is activated by PPARA and induced by phytanic acid and fibrates. The downstream metabolites pristanal and pristanic acid are processed by AMACR for beta-oxidation, while formyl-CoA is hydrolyzed. HACL1 requires PEX5 for peroxisomal import and TPP as a cofactor. The pathway operates downstream of PHYH, which hydroxylates phytanoyl-CoA. Disruption of HACL1 causes 2-hydroxyphytanoyl-CoA accumulation and blocks pristanal and formyl-CoA production, phenocopying Refsum disease.
In the SK-HEP-1 hepatocellular carcinoma context, HACL1 knockout generates a model of peroxisomal alpha-oxidation deficiency superimposed on a cancer metabolic background. This allows dissection of how impaired phytanic acid degradation influences cancer cell behavior, potentially revealing synthetic lethalities or adaptive metabolic shifts. Moreover, the model is valuable for studying peroxisome?Ccancer crosstalk, as peroxisomal metabolism contributes to ROS homeostasis, bile acid synthesis, and energy metabolism??all pathways often rewired in liver tumors.
This polyclonal knockout cell product is suited for modeling Refsum disease, peroxisomal biogenesis disorders, and lipid metabolism dysregulation in liver cancer. Typical assays include western blotting and RT-qPCR for HACL1 validation, phytanic acid accumulation assays to confirm metabolic blockade, immunofluorescence staining of peroxisomal markers (e.g., PEX5, catalase), and metabolic labeling with phytanic acid to trace alpha-oxidation flux. These cells also enable drug screening for compounds that restore peroxisomal function or mitigate lipid toxicity. For further information, contact Ascent Research.
