HSD17B4 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the HSD17B4 gene is disrupted. This pooled format consists of a heterogeneous collection of edited cells each carrying distinct mutations around the target site, yielding a loss-of-function model. The product provides a versatile tool for investigating HSD17B4 deficiency in a near-haploid human background.
The HAP1 host cell line is a near-haploid human male adherent line derived from the KBM-7 chronic myeloid leukemia (CML) line. Its near-haploid karyotype reduces genetic complexity, facilitating unambiguous gene disruption studies. HAP1 cells are p53-deficient, which eliminates confounding DNA damage responses and makes them particularly suitable for metabolic and genotoxic stress assays.
HSD17B4 encodes a peroxisomal bifunctional enzyme catalyzing the hydration and dehydrogenation of enoyl-CoA during ??-oxidation of fatty acids. This enzyme operates downstream of acyl-CoA oxidase 1 (ACOX1) and works in concert with sterol carrier protein 2 (SCP2) and acetyl-CoA acyltransferase 1 (ACAA1) to complete the shortening of fatty acyl chains, generating acetyl-CoA and NADH. Its import into peroxisomes relies on the receptors PEX5 and PEX7. Transcriptional regulation by PPAR??, PPAR??, RXR, and THR couples HSD17B4 expression to lipid metabolism and peroxisomal biogenesis. Additionally, HSD17B4 participates in steroid hormone metabolism, interconverting estrone and testosterone, thereby influencing steroid profiles.
In the HAP1 background, the HSD17B4 knockout recapitulates biochemical hallmarks of D-bifunctional protein deficiency and Zellweger spectrum disorders, including accumulation of very long-chain fatty acids (VLCFAs) and disrupted steroid hormone levels. The near-haploid genome ensures complete loss of function from a single allele, while the p53 deficiency avoids confounding apoptotic responses. This model thus provides a clean genetic system to dissect peroxisomal ??-oxidation pathways and their interplay with steroid biosynthesis, enabling robust mechanistic studies.
This polyclonal knockout population is ideal for applications such as peroxisomal disorder modeling, fatty acid metabolism flux studies, and drug screening for peroxisome-associated diseases. Researchers can validate functional consequences using assays including Western blotting for protein expression, RT-qPCR for transcriptional effects, and lipidomic analysis of VLCFAs. Immunofluorescence microscopy can assess peroxisomal morphology, while LC-MS/MS quantification of steroid hormones enables endocrine profiling. Cell viability under oxidative stress provides a readout for peroxisomal function. The heterogeneous knockout pool is also well-suited for CRISPR functional screens. For further details, contact Ascent Research.
