The ACBD5 knockout HeLa polyclonal cells are a CRISPR/Cas9-edited heterogeneous cell population derived from the HeLa cervical adenocarcinoma line, designed for studies of peroxisome biology and membrane contact sites. This polyclonal pool contains multiple disrupted ACBD5 alleles, generating a loss-of-function model that avoids clonal artifacts. The product enables robust interrogation of ACBD5-dependent pathways without the need for single-cell clone isolation.
The parental HeLa cell line is an immortalized human cervical epithelial adenocarcinoma model, positive for HPV18, and is widely used in cancer and cell biology research. HeLa cells offer a well-characterized genetic background, reliable culture characteristics, and retain key organelle systems including peroxisomes and ER, making them suitable for knockout-based studies of membrane contact site and lipid metabolism pathways.
ACBD5 is a peroxisomal membrane protein that acts as a critical mediator of ER-peroxisome contact sites through direct interaction with the ER-localized VAPA and VAPB proteins. This tethering complex facilitates the exchange of phospholipids and metabolites essential for peroxisome biogenesis and very long-chain fatty acid (VLCFA) metabolism. ACBD5 functions downstream of putative regulators such as PPAR?? and peroxisome proliferators, and it is required for efficient peroxisomal beta-oxidation, positioning it within a pathway that includes ACOX1, DBP, and various PEX proteins. Knockout of ACBD5 disrupts these contact sites, leading to impaired VLCFA breakdown, altered peroxisomal homeostasis, and defective phospholipid transfer.
In the HeLa cancer cell context, ACBD5 knockout provides a platform to explore how peroxisome-ER contacts influence lipid metabolic reprogramming and organelle dynamics in tumor cells. As ACBD5 is also linked to peroxisomal disorders, retinal dystrophy, and leukodystrophy, this model can be used to dissect disease-relevant mechanisms and evaluate potential therapeutic interventions targeting membrane contact sites in a human cell environment.
This polyclonal knockout is suitable for diverse applications, including the study of ER-peroxisome membrane contact site formation, very long-chain fatty acid metabolic pathways, and modeling of peroxisomal dysfunction. Recommended assays include immunofluorescence labeling of peroxisomal markers such as PMP70, Western blotting for ACBD5 and VAP proteins, co-immunoprecipitation of VAPA/VAPB complexes, GC-MS-based VLCFA analysis, and peroxisomal beta-oxidation activity measurements. For additional information, please contact Ascent Research.
