The HACL2 Knockout A-549 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of A-549 lung adenocarcinoma epithelial cells carrying targeted disruption of the HACL2 gene, which encodes peroxisomal 2-hydroxyacyl-CoA lyase. This polyclonal format provides a heterogeneous pool of knockout cells, minimizing clonal bias and enabling robust population-level studies of HACL2 loss-of-function. The use of CRISPR/Cas9-mediated gene disruption ensures efficient target ablation, and the polyclonal nature allows investigation of gene function without the limitations of single-cell-derived clones.
The parental A-549 cell line originates from human lung adenocarcinoma and harbors a KRAS G12S mutation with wild-type p53, representing a classic model for KRAS-driven non-small cell lung cancer. Its epithelial morphology and well-documented genetic background support research on metabolism, signaling, and drug resistance. This context is particularly relevant for studying how peroxisomal alpha-oxidation pathways influence lung adenocarcinoma biology.
HACL2 functions as a TPP- and Mg2+-dependent lyase that cleaves 2-hydroxy fatty acyl-CoAs into formyl-CoA and fatty aldehydes during peroxisomal alpha-oxidation. The enzyme acts downstream of PHYH, which hydroxylates phytanoyl-CoA to produce 2-hydroxyphytanoyl-CoA. The resulting fatty aldehyde is oxidized by ALDH3A2 to pristanic acid, a step critical for branched-chain fatty acid degradation. HACL2 activity is regulated by PPARalpha, and its peroxisomal import requires PEX5. Knockout of HACL2 consequently blocks alpha-oxidation, leading to accumulation of phytanic acid and 2-hydroxy fatty acyl-CoAs, which can disrupt mitochondrial function, induce oxidative stress, and perturb lipid homeostasis.
In A-549 lung adenocarcinoma cells, HACL2 knockout impairs peroxisomal alpha-oxidation, likely resulting in toxic accumulation of phytanic acid and 2-hydroxy fatty acyl-CoA intermediates. This metabolic disruption may sensitize cells to oxidative stress and alter KRAS-driven lipid metabolic reprogramming, potentially affecting proliferation and survival. The model enables dissection of the crosstalk between peroxisomal function and oncogenic signaling, providing insight into how aberrant fatty acid metabolism impacts lung adenocarcinoma pathology and drug response.
This polyclonal knockout cell population is suited for investigating cancer lipid metabolism and peroxisomal disorders, with applications including phytanic acid challenge assays coupled with GC-MS, Western blotting and RT-qPCR for HACL2 expression, metabolic flux analysis, and functional assays such as colony formation and migration. Its use facilitates studies on alpha-oxidation-related toxicity, drug sensitivity profiling, and the role of 2-hydroxy fatty acid metabolism in lung adenocarcinoma progression. For further product information or technical support, please contact Ascent Research.
