HSD17B8 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt HSD17B8 in the human near-haploid HAP1 leukemic cell line. This loss-of-function model enables dissection of the gene??s dual enzymatic activities: 17??-hydroxysteroid dehydrogenase?Cmediated steroid hormone biosynthesis and 3-ketoacyl-CoA reductase function in mitochondrial long-chain fatty acid elongation. The polyclonal format comprises a heterogeneous mixture of edited cells, reflecting diverse mutational outcomes and providing a robust platform for functional genomics and metabolic studies.
The HAP1 cell line is a male, adherent, near-haploid line derived from the chronic myeloid leukemia KBM-7. Its near-haploid karyotype facilitates efficient CRISPR/Cas9 gene disruption, making it a preferred model for knockout experiments. HAP1 retains core metabolic pathways and oncogenic signaling networks, including steroidogenic and fatty acid elongation cascades, rendering it an appropriate host for investigating hormone-dependent cancers and inborn metabolic defects.
HSD17B8 encodes a bifunctional enzyme. As a 17??-hydroxysteroid dehydrogenase, it reduces 17-ketosteroids such as androstenedione to testosterone; as a 3-ketoacyl-CoA reductase, it participates in mitochondrial fatty acid elongation. Transcription is driven by steroidogenic factor 1 (SF1/NR5A1) and PPAR??. The protein physically interacts with HSD17B12 and TECR and cooperates with ELOVL6 to generate very long-chain fatty acids. Consequently, HSD17B8 disruption alters downstream estradiol and testosterone levels, as well as very long-chain fatty acid profiles, impacting endocrine and lipid homeostasis.
Knockout of HSD17B8 in HAP1 cells perturbs steroid hormone biosynthesis and fatty acid metabolism, creating a disease-relevant model for polycystic ovary syndrome, HSD17B8 deficiency, and hormone-dependent malignancies. The leukemic background allows assessment of steroid-driven cancer cell proliferation and apoptosis, while the polyclonal nature captures biological heterogeneity. These cells also facilitate exploration of mitochondrial dysfunction in metabolic disorders.
Typical experimental approaches include western blotting and RT-qPCR to quantify steroidogenic enzyme expression, liquid chromatography?Cmass spectrometry (LC-MS) for steroid profiling, and gas chromatography?Cmass spectrometry for fatty acid composition. Functional assays such as hormone-stimulated proliferation, steroid receptor luciferase reporters, and immunofluorescence for mitochondrial localization further validate pathway engagement. The polyclonal population is suited for high-throughput drug screening targeting endocrine and metabolic pathways. For detailed protocols, please contact Ascent Research.
