The ACOX1 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for the disruption of the ACOX1 gene in a widely utilized human epithelial model. This polyclonal product provides a heterogeneous pool of cells carrying diverse gene-disruption events, enabling robust functional studies without the need for single-cell clonal isolation. The use of CRISPR/Cas9 technology ensures efficient target-gene disruption while maintaining the genetic background of the host cell line.
HeLa cells, derived from a cervical adenocarcinoma of a 31-year-old female, are one of the most extensively characterized immortalized human cell lines. They harbor integrated human papillomavirus 18 (HPV-18) DNA, and the viral oncoproteins E6 and E7 inactivate the tumor suppressors p53 and Rb, respectively. These features confer unlimited proliferative capacity and have made HeLa cells a fundamental platform for research in cancer biology, virology, signal transduction, and drug discovery.
ACOX1 (acyl-CoA oxidase 1) catalyzes the initial and rate-limiting step of peroxisomal very long-chain fatty acid (VLCFA) ??-oxidation, converting acyl-CoA substrates into trans-2-enoyl-CoA with concomitant generation of hydrogen peroxide (H?O?). The enzyme is imported into peroxisomes via an interaction network involving PEX5, PEX14, and PEX13. ACOX1 expression is under transcriptional control of PPAR??, PPAR??, thyroid hormone receptor, glucocorticoid receptor, and insulin signaling. Its activity produces HSD17B4 and SCPx substrates, and contributes to the formation of acetyl-CoA and PPAR??-activating lipid ligands, thereby linking peroxisomal metabolism to broader metabolic regulation and reactive oxygen species signaling.
In the HeLa cell context, CRISPR/Cas9-mediated ACOX1 knockout disrupts peroxisomal VLCFA catabolism, leading to intracellular accumulation of very long-chain fatty acids and a reduction in peroxisomally derived H?O?. Given the transformed metabolic state of HeLa cells and the role of lipid metabolism in supporting cancer cell proliferation and survival, this knockout model provides a valuable system to dissect the interplay between peroxisomal function, oxidative stress, and oncogenic signaling. It also serves as an in vitro surrogate for peroxisomal biogenesis disorders such as neonatal adrenoleukodystrophy and Zellweger spectrum disorder.
This polyclonal knockout product is well-suited for applications in lipid metabolism, oxidative stress, and cancer metabolism research. Typical assays include Western blotting for ACOX1 protein levels, very long-chain fatty acid quantification by GC-MS, H?O? measurement, peroxisomal ??-oxidation activity assays, immunofluorescence staining of peroxisomal markers, and transcriptional profiling by RT-qPCR or RNA-seq. The cells may also be employed in drug toxicity screens targeting peroxisomal pathways. For further details or technical support, please contact Ascent Research.
