HSDL1 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the HSDL1 gene in the HAP1 near-haploid human cell line. HSDL1 encodes a peroxisomal 2-hydroxyacyl-CoA dehydrogenase that catalyzes the NAD+-dependent oxidation of 2-hydroxy fatty acyl-CoAs to 2-ketoacyl-CoAs, a central step in the peroxisomal alpha-oxidation pathway. This product provides a robust loss-of-function model for investigating the molecular basis of peroxisomal fatty acid metabolism and related disorders.
The HAP1 host cell line is a near-haploid human cell model derived from the KBM-7 chronic myeloid leukemia line. HAP1 retains a single copy of most chromosomes, thereby eliminating the confounding effects of diploid heterozygosity and enabling unambiguous genotype-phenotype correlations. This feature makes HAP1 an ideal platform for genetic knockout screens and functional genomics studies, particularly for genes involved in metabolic pathways where allele dosage can mask subtle phenotypes.
HSDL1, a peroxisomal 2-hydroxyacyl-CoA dehydrogenase, catalyzes the NAD+-dependent oxidation of 2-hydroxy fatty acyl-CoAs to 2-ketoacyl-CoAs within the alpha-oxidation pathway. It functions downstream of phytanoyl-CoA hydroxylase (PAHX) and 2-hydroxyphytanoyl-CoA lyase, and its products are further metabolized by peroxisomal acyl-CoA oxidases. Interaction with the import receptor PEX5 mediates peroxisomal targeting, while transcription is regulated by the nuclear receptors PPAR-alpha and PPAR-gamma in response to fatty acid ligands. Disruption of HSDL1 thus disrupts this catalytic sequence, leading to accumulation of 2-hydroxy fatty acids and impaired degradation of branched-chain fatty acids such as phytanic acid.
The near-haploid HAP1 background eliminates confounding diploid heterozygosity, ensuring uniform loss of HSDL1 function across the polyclonal population and simplifying phenotypic analyses. This model is especially valuable for studying peroxisomal disorders, fatty acid oxidation defects, and metabolic syndrome, where HSDL1 disruption may contribute to pathogenic lipid accumulation and aberrant peroxisomal function.
These polyclonal knockout cells are suitable for diverse applications, including western blotting for HSDL1 confirmation, RT-qPCR profiling of peroxisomal gene expression, and functional fatty acid oxidation assays. Lipidomics and metabolomics can characterize 2-hydroxy fatty acid accumulation, while immunofluorescence imaging assesses peroxisome integrity. The model supports drug testing for compounds that may compensate for HSDL1 deficiency, providing a platform for therapeutic discovery in peroxisomal and metabolic diseases. For further information, please contact Ascent Research.
