The ACBD5 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population targeting the ACBD5 gene in the HT29 colorectal adenocarcinoma cell line. This model provides a heterogeneous pool of cells with disrupted ACBD5 expression, enabling loss-of-function analysis without monoclonal selection. The gene encodes a peroxisomal membrane protein essential for tethering peroxisomes to the endoplasmic reticulum (ER), a process critical for lipid exchange and metabolic regulation.
HT29 cells are an adherent epithelial line isolated from a 44-year-old female with colorectal adenocarcinoma. These cells retain the capacity for enterocytic differentiation, making them a well-characterized intestinal epithelial model. Their robust growth and documented use in cancer metabolism research provide a relevant context for investigating peroxisomal biology in colorectal tumors.
ACBD5 functions as a peroxisomal anchor that interacts with the ER-resident proteins VAPB and VAPA, forming membrane contact sites that facilitate transfer of very long-chain fatty acids for ??-oxidation. The protein is regulated upstream by the transcription factor PPAR?? and fatty acid ligands. Downstream signaling involves enzymes such as ACOX1 and HSD17B4, which execute fatty acid catabolism. Disruption of ACBD5 therefore impairs peroxisome-ER tethering and downstream ??-oxidation processes.
In the HT29 colorectal adenocarcinoma background, ACBD5 knockout provides a valuable system to study the contribution of peroxisomal lipid metabolism to cancer cell proliferation. Colorectal tumors often rewire metabolic pathways to sustain growth, and intact peroxisome-ER contacts may support the elevated fatty acid oxidation required for tumor maintenance. This polyclonal knockout model allows dissection of how loss of ACBD5 alters metabolic homeostasis, cell viability, and potentially differentiation in intestinal epithelial-derived cancer cells.
Applications include immunofluorescence for peroxisomal markers (e.g., PMP70), co-immunoprecipitation of VAPB, and fatty acid oxidation assays. Lipidomics and RT-qPCR profiling can characterize metabolic and transcriptional changes. These cells are also suitable for drug screening in peroxisomal disorder research and cell viability assays under metabolic stress. For further information, please contact Ascent Research.
