The EHHADH Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human A-549 lung adenocarcinoma cell line, featuring disruption of the EHHADH gene. This polyclonal pool captures a range of editing events, enabling robust functional studies while preserving natural genetic heterogeneity. The knockout model is generated without single-cell cloning, making it suitable for experiments where population-level effects are desired. The cells provide a loss-of-function platform to dissect peroxisomal fatty acid beta-oxidation pathways in an epithelial context.
The parental A-549 cell line was originally isolated from a 58-year-old Caucasian male with lung adenocarcinoma. These cells exhibit an epithelial morphology and are widely utilized in cancer biology, respiratory disease research, and toxicology. Their well-characterized growth properties and responsiveness to metabolic and pharmacological interventions make them a practical host for genetic perturbation studies, particularly those examining lipid metabolism and peroxisomal function in tumor cells.
EHHADH encodes the L-bifunctional protein, a peroxisomal enzyme catalyzing the second and third steps of fatty acid beta-oxidation: hydration of trans-2-enoyl-CoA and dehydrogenation of 3-hydroxyacyl-CoA. Its expression is transcriptionally regulated by PPAR??, which heterodimerizes with RXR?? and is activated by ligands such as fatty acids and fibrates, with additional input from HNF4??. The enzyme functions within a peroxisomal complex that includes PEX5-mediated import, interacting with SCP2 and acyl-CoA oxidase, and contributes to the production of acetyl-CoA and medium-chain acyl-CoA that feed downstream pathways including lipid metabolism and energy homeostasis.
In A-549 lung cancer cells, EHHADH knockout disrupts peroxisomal degradation of long-chain and branched-chain fatty acids, potentially shifting cellular lipid metabolism and altering energy homeostasis. Given the reliance of tumor cells on metabolic flexibility for proliferation and survival, this model is significant for investigating peroxisome-dependent metabolic reprogramming in lung adenocarcinoma. The knockout may also impact ROS balance and peroxisome-mediated signaling, offering insights into the interplay between peroxisomal function and cancer cell physiology.
This polyclonal knockout cell population is well-suited for a variety of research applications, including measurement of fatty acid oxidation flux using isotope-labeled substrates, lipidomic profiling to assess changes in very long-chain fatty acids and related metabolites, and immunofluorescence analysis of peroxisomal markers. It also supports drug sensitivity screening and functional assays exploring the role of peroxisomes in drug resistance mechanisms. Researchers can employ these cells to model peroxisomal dysfunction relevant to metabolic diseases such as Fanconi syndrome. For further information and expert support, please contact Ascent Research.
