The HACD3 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HeLa human cervical adenocarcinoma epithelial cell line. This polyclonal pool carries targeted disruption of HACD3, encoding 3-hydroxyacyl-CoA dehydratase 3, a critical enzyme in the very-long-chain fatty acid (VLCFA) elongation cycle. The polyclonal format minimizes clonal selection bias, providing a robust loss-of-function model for studying HACD3-dependent lipid metabolism.
HeLa cells, originally established from a cervical adenocarcinoma biopsy in 1951, are immortalized by integrated HPV-18 and are widely utilized in cancer research, virology, and cell biology. Their transformed phenotype includes altered lipid metabolism, making them an appropriate host for investigating enzymes involved in VLCFA and sphingolipid synthesis.
HACD3 catalyzes the dehydration of 3-hydroxyacyl-CoA to trans-2-enoyl-CoA, the third step of the ER-resident VLCFA elongase complex that also contains ELOVL elongases, HACD family members, TECR, and ACSL1. Transcription of HACD3 is regulated by SREBP1, PPAR??, and LXR. The VLCFAs produced are essential precursors for ceramide, sphingomyelin, and lipid droplet formation. Thus, HACD3 sits at an intersection of fatty acid elongation and sphingolipid biosynthesis, with broad impacts on membrane structure and signaling.
In the HeLa carcinoma background, HACD3 disruption enables dissection of how oncogenic pathways interface with lipid homeostasis. VLCFA-derived sphingolipids influence membrane microdomain organization and lipid droplet dynamics, processes frequently dysregulated in cancer. Additionally, given that HACD3 mutations are linked to neurodevelopmental disorders including intellectual disability, developmental and epileptic encephalopathy, and microcephaly, these polyclonal knockout cells provide a platform for exploring the metabolic underpinnings of such conditions when paired with disease-relevant cellular contexts.
Researchers can employ this model in fatty acid elongation assays, lipidomics (LC-MS), ceramide ELISA, BODIPY staining, and immunofluorescence to dissect VLCFA and sphingolipid metabolism. Expression analysis of regulatory factors such as SREBP1 and PPAR?? via RT-qPCR or Western blotting is also supported. The HACD3 Knockout HeLa Polyclonal Cells thus offer a versatile tool for both targeted lipid studies and broader metabolic investigations. For technical information, contact Ascent Research.
