The HACD2 Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the haploid HAP1 cell line, with targeted disruption of the HACD2 gene. This product represents a heterogeneous pool of cells harboring HACD2 gene disruptions achieved via CRISPR/Cas9 technology, providing a robust loss-of-function model for studying the enzymatic step catalyzed by 3-hydroxyacyl-CoA dehydratase 2 in very long-chain fatty acid elongation.
HAP1 cells are a near-haploid human cell line originally isolated from the KBM-7 chronic myeloid leukemia background. Their haploid karyotype simplifies genetic studies by facilitating the generation of single-gene disruptions and reducing the complexity of diploid gene redundancy, making them an ideal host for knockout modeling. HAP1 cells retain expression of many metabolic pathways and are widely employed in functional genomics screens, drug target validation, and pathway dissection.
HACD2 encodes the third enzyme of the microsomal fatty acid elongation cycle, specifically catalyzing the dehydration of 3-hydroxyacyl-CoA intermediates to trans-2-enoyl-CoA during the elongation of very long-chain fatty acids (VLCFAs; C16?CC26). This reaction operates immediately downstream of the ELOVL family of elongases and upstream of trans-2,3-enoyl-CoA reductase (TECR, also known as TER). HACD2 is transcriptionally regulated by lipogenic factors including SREBP1c and PPAR??, and its activity integrates insulin signaling with lipid anabolism. Interacting partners include the ELOVL1-7 elongases, the terminal reductase TECR, and ER membrane complex proteins that organize the elongase complex. Disruption of HACD2 leads to accumulation of 3-hydroxyacyl-CoA species, reduced production of VLCFAs, and downstream perturbations in the synthesis of sphingolipids and ceramides.
In the HAP1 haploid background, the knockout of HACD2 creates a clean metabolic perturbation that unmasks the cellular reliance on dehydratase activity for maintaining VLCFA pools. The polyclonal population ensures representation of multiple gene-disrupted alleles, minimizing clonal artifacts and providing a physiologically broad knockout phenotype. This model is particularly valuable for investigating lipid remodeling under conditions where VLCFA elongation is compromised, such as in metabolic syndromes or certain neuropathies linked to altered fatty acid metabolism.
This knockout cell pool is suited for a wide range of functional studies, including lipidomic profiling by LC-MS to quantify VLCFA and sphingolipid species, FAME analysis to assess fatty acid composition, and RT-qPCR or western blotting to monitor compensatory changes in elongation pathway components. It can also be applied in drug screening campaigns aimed at restoring or bypassing the dehydration step, and in CRISPR-based genetic interaction studies to map functional relationships within the elongase complex. Researchers interested in further details or bulk pricing are encouraged to contact Ascent Research.
