The HACL1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the HACL1 gene in the human HAP1 cell line. This gene-targeting approach yields a loss-of-function model for interrogating peroxisomal alpha-oxidation and the metabolic consequences of HACL1 ablation. The polyclonal format provides a diverse array of edited alleles, enabling robust phenotypic analysis at the population level.
The host HAP1 cell line is a near-haploid human chronic myeloid leukemia line derived from KBM-7. Its minimal karyotype, with only a partial disomy on chromosome 15, simplifies functional genomics studies by reducing genetic redundancy and facilitating complete gene inactivation. HAP1 cells are a gold standard for CRISPR-based screens and genetic interaction mapping, offering a clean background for dissecting HACL1 function.
HACL1 encodes 2-hydroxyphytanoyl-CoA lyase, a peroxisomal enzyme that catalyzes the carbon-carbon bond cleavage of 2-hydroxyacyl-CoA thioesters, generating formyl-CoA and a fatty aldehyde. This step is central to the alpha-oxidation pathway for phytanic acid degradation. HACL1 operates downstream of phytanoyl-CoA hydroxylase (PHYH) and upstream of aldehyde dehydrogenase ALDH3A2. Its catalytic activity depends on peroxisomal import mediated by PEX proteins and the cofactor coenzyme A. The gene is transcriptionally regulated by PPAR signaling and transcription factors governing peroxisome biogenesis, placing HACL1 within a network that links peroxisomal fatty acid metabolism to cellular lipid homeostasis.
Disruption of HACL1 in HAP1 cells impairs phytanic acid alpha-oxidation, leading to the accumulation of phytanic acid and its CoA esters, a phenotype reminiscent of peroxisomal disorders such as Refsum disease. This polyclonal knockout model enables detailed metabolic tracing and genetic interaction studies, and the near-haploid background ensures strong loss-of-function effects without compensatory wild-type alleles.
Experimental applications include fatty acid oxidation assays, phytanic acid accumulation measurements, peroxisomal import assays, immunoblotting for HACL1, and RT-qPCR analysis of peroxisomal gene expression. The cells are well-suited for CRISPR functional genomics, drug sensitivity profiling, and mechanistic studies of peroxisomal disorders. For further details, please contact Ascent Research.
