The ABCD1 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 human hepatocellular carcinoma cell line, featuring targeted disruption of the ABCD1 gene. This polyclonal format offers a heterogeneous pool of edited cells with stable ABCD1 loss-of-function, ideal for studying peroxisomal fatty acid metabolism and related disease mechanisms. Unlike clonal lines, the population approach provides a broader representation of CRISPR-driven genetic heterogeneity, making it suitable for pooled functional screens and robust phenotypic analyses.
The Huh-7 cell line is an epithelial hepatoma model widely used in liver biology and metabolism research. Derived from a human hepatocellular carcinoma, Huh-7 cells retain many hepatocyte-like characteristics, including expression of liver-specific metabolic enzymes and active lipid metabolism pathways. Their robust proliferation and well-characterized genome make them a valuable platform for dissecting hepatic peroxisomal functions and investigating the cellular consequences of lipid transporter defects in a liver epithelial context.
ABCD1 encodes the peroxisomal ATP-binding cassette transporter ALDP, which mediates the uptake of very-long-chain fatty acyl-CoAs into peroxisomes for ??-oxidation. Its expression is transcriptionally regulated by the nuclear receptor PPAR?? in response to fatty acid ligands. ALDP interacts with related transporters ABCD2 and ABCD3, and its peroxisomal targeting depends on the chaperone PEX19. Additionally, ALDP cooperates with acyl-CoA synthetases to channel substrates into the organelle. Disruption of ABCD1 blocks substrate import, leading to cytosolic accumulation of very-long-chain fatty acids and impaired peroxisomal ??-oxidation, with downstream effects on enzymes such as ACOX1 and DBP.
In Huh-7 hepatocellular carcinoma cells, ABCD1 knockout compromises peroxisomal lipid handling, causing abnormal very-long-chain fatty acid accumulation and potential oxidative stress. This phenotype recapitulates key metabolic defects observed in X-linked adrenoleukodystrophy and related peroxisomal disorders, making the model valuable for investigating disease mechanisms in a hepatic context. Moreover, the disruption may intersect with oncogenic lipid metabolism and redox homeostasis, offering insights into liver cancer biology where peroxisomal functions are often dysregulated.
Researchers can employ this polyclonal knockout population to explore very-long-chain fatty acid metabolism, peroxisome biogenesis, and the molecular pathogenesis of peroxisomal disorders. Representative assays include western blotting and RT-qPCR for ABCD1, LC-MS-based quantification of very-long-chain fatty acids, peroxisomal ??-oxidation assays, immunofluorescence for peroxisomal markers such as PMP70, and cellular lipidomics. The model is also suited for drug testing and viability assays under lipid overload. For further inquiries, please contact Ascent Research.
