The HACL1 Knockout A-549 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population for targeted disruption of the HACL1 gene in human A-549 cells. This loss-of-function model enables investigation of 2-hydroxyacyl-CoA lyase function and alpha-oxidation pathway dynamics. The polyclonal pool retains diverse edits across the population, offering a robust system for biochemical and functional assays in a gene knockout context without reliance on single-cell clones.
A-549 cells are an adherent epithelial line originally isolated from a 58-year-old Caucasian male with lung adenocarcinoma, displaying typical epithelial morphology. They serve as a well-characterized in vitro model of human alveolar type II epithelial cells, retaining key metabolic features. Their tumorigenic origin and active peroxisomal pathways make them especially suitable for exploring the intersection of cancer metabolism and peroxisomal lipid processing.
HACL1 encodes a peroxisomal 2-hydroxyacyl-CoA lyase that catalyzes the cleavage of 2-hydroxyacyl-CoA into formyl-CoA and a fatty aldehyde, a critical step in the alpha-oxidation of phytanic acid and branched-chain fatty acid degradation. This enzyme is transcriptionally regulated by PPARA and depends on PEX5-directed import via interaction with PEX14 at the peroxisomal importomer. Downstream, the aldehyde product is oxidized by ALDH3A2, formyl-CoA hydrolase processes the formyl-CoA, and pristanic acid generated is further metabolized by SCP2. The pathway, initiated by PHYH, ensures efficient clearance of potentially toxic branched-chain lipids.
Knockout of HACL1 in A-549 cells blocks alpha-oxidation, leading to phytanic acid and 2-hydroxyacyl-CoA accumulation, mirroring aspects of Refsum disease and peroxisomal biogenesis disorders. This model enables dissection of peroxisomal dysfunction and investigation of how impaired branched-chain fatty acid degradation influences cancer metabolic reprogramming, lipid homeostasis, and epithelial cell biology.
Researchers can apply this model to disease modeling, drug screening, and metabolic flux studies. Reproducible assays include LC-MS/MS-based phytanic acid quantification, PMP70 immunofluorescence for peroxisomal visualization, RT-qPCR and Western blotting for gene/protein analysis, and fatty acid oxidation assays. The system supports investigations into Refsum disease, peroxisomal biogenesis disorders, and the contribution of alpha-oxidation to lung adenocarcinoma metabolic reprogramming. For further information, please contact Ascent Research.
