The ABCD1 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HeLa cells, designed for loss-of-function studies of the human ABCD1 gene. This model uses CRISPR/Cas9-mediated gene disruption across the cell pool, avoiding clonal artifacts and enabling flexible interrogation of peroxisomal metabolism. It is suited for research into X-linked adrenoleukodystrophy (X-ALD) and very-long-chain fatty acid (VLCFA) biology in a human epithelial line.
HeLa cells, an HPV18-positive cervical adenocarcinoma epithelial line, serve as a robust host due to rapid growth, genetic manipulability, and well-documented cancer biology. Their transformed status offers a convenient platform for studying peroxisome-related processes, despite potential differences from primary cells. HeLa’s tractability supports high-throughput CRISPR editing and live-cell imaging assays, making them useful for exploring ABCD1-dependent functions.
ABCD1 encodes ALDP, a peroxisomal ABC transporter that imports VLCFA-CoA esters, especially C26:0, for ??-oxidation by ACOX1, DBP, and SCPx. Its expression is transcriptionally regulated by PPARA, PPARG, RXR, and LXR in response to lipid ligands. ALDP interacts with peroxins PEX19, PEX3, PEX16, and forms homodimers, while also cooperating with ABCD2 and ABCD3. Disruption impairs VLCFA degradation, causing accumulation, oxidative stress, and downstream defects in bile acid, DHA, and plasmalogen synthesis, mirroring X-ALD metabolic derangements.
In HeLa cells, ABCD1 knockout recapitulates X-ALD features by elevating C26:0 and triggering peroxisomal dysfunction. This polyclonal model captures heterogeneous gene disruption, reflecting population-level effects and complementing clonal lines. It enables examination of VLCFA lipotoxicity, peroxisome-mitochondria crosstalk, and redox imbalance relevant to adrenomyeloneuropathy and Addison disease. The model supports mechanistic studies linking transporter loss to metabolic and oxidative stress pathways.
Key applications include X-ALD disease modeling with LC-MS/MS-based C26:0 quantification, western blotting for ABCD1, and immunofluorescence for peroxisomal markers. The cells are suitable for screening therapies to reduce VLCFA levels, evaluating small molecules, and conducting RNA-seq transcriptome profiling. Additional assays like fatty acid ??-oxidation measurement, ROS detection, and MTT viability allow multi-parametric analysis. For further information or technical inquiries, contact Ascent Research.
