The HACD3 Knockout HAP1 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 cell line, featuring a targeted disruption of the human HACD3 gene. This loss-of-function model enables investigation of very long-chain fatty acid (VLCFA) elongation and its downstream consequences in a near-haploid genetic background.
HAP1 cells are a chronic myeloid leukemia-derived, near-haploid hematopoietic cell line with an adherent, fibroblast-like morphology. Originally adapted from the KBM-7 CML line, HAP1 cells provide a simplified genetic landscape ideal for knockout-based functional genomics studies, as the near-haploid genome reduces functional redundancy and facilitates unambiguous genotype?Cphenotype analyses.
HACD3 encodes 3-hydroxyacyl-CoA dehydratase 3, which catalyzes the dehydration step of the VLCFA elongation cycle in the endoplasmic reticulum. The enzyme functions downstream of the ELOVL elongases (ELOVL1, ELOVL2, ELOVL3) and malonyl-CoA-dependent chain extension, acting in concert with trans-2,3-enoyl-CoA reductase (TECR) and 17??-hydroxysteroid dehydrogenase 12 (HSD17B12) to generate very long-chain acyl-CoAs such as behenoyl-CoA and lignoceroyl-CoA. HACD3 activity is transcriptionally regulated by SREBP1 in response to insulin signaling and palmitoyl-CoA substrate availability. The resulting VLCFA-CoAs serve as critical precursors for sphingolipid and ceramide biosynthesis. Disruption of HACD3 therefore impairs the production of these lipid species, potentially altering membrane architecture and lipid-mediated signal transduction.
In the HAP1 leukemia background, HACD3 knockout provides a unique platform to dissect the role of VLCFA metabolism in hematological malignancy. Aberrant lipid metabolism is increasingly recognized as a hallmark of cancer, and HACD3-dependent VLCFA synthesis may contribute to membrane biogenesis, lipid raft assembly, and survival signaling in leukemia cells. This model thus allows researchers to examine how VLCFA dysregulation influences cellular proliferation, differentiation, and sensitivity to therapeutic agents, while also offering relevance to VLCFA synthesis defects and associated neurodevelopmental disorders.
Research applications include quantitative lipidomics by LC?CMS to profile VLCFA species and sphingolipid alterations, immunoblotting and RT-qPCR to assess expression of fatty acid elongation enzymes, cell proliferation and viability assays under varying lipid conditions, and radiolabeled fatty acid elongation activity assays to directly measure pathway flux. The polyclonal population can be employed in pooled screening campaigns to identify modulators of fatty acid elongation or to uncover synthetic lethal interactions in leukemia cells. For additional technical details or to request a quote, please contact Ascent Research.
