The HSD17B10 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population targeting the HSD17B10 gene, which encodes a mitochondrial short-chain dehydrogenase/reductase. This heterogeneous pool of HAP1 cells carries diverse loss-of-function mutations in HSD17B10, eliminating the need for clonal selection and enabling bulk analyses of gene disruption effects.
HAP1 is a near-haploid human cell line derived from the chronic myeloid leukemia cell line KBM-7. The near-haploid karyotype ensures that a single CRISPR-mediated mutation can yield functional gene inactivation, simplifying functional genomics studies. HAP1 cells maintain core cellular machinery, making them suitable for investigating mitochondrial metabolism, signal transduction, and disease pathways.
HSD17B10 catalyzes the oxidation of 17??-estradiol to estrone and is essential for branched-chain amino acid degradation, converting isovaleryl-CoA to 2-methyl-3-hydroxybutyryl-CoA. The enzyme binds amyloid-beta peptide (A??) and interacts with mitochondrial complex I subunits and Cyclophilin D, linking it to energy metabolism and apoptosis. Upstream regulators include substrate availability and PGC-1??-mediated mitochondrial biogenesis. Downstream, HSD17B10 influences estrone levels, 2-methyl-3-hydroxybutyryl-CoA production, reactive oxygen species, and mitochondrial respiratory chain activity.
In the HAP1 context, HSD17B10 knockout recapitulates key biochemical features of HSD10 mitochondrial disease and disrupts A??-associated mitochondrial toxicity, providing a model for Alzheimer??s disease research. The loss of enzymatic activity leads to substrate accumulation and impaired mitochondrial fatty acid ??-oxidation. The haploid background facilitates clear genotype-phenotype correlations, enhancing phenotypic characterization.
These polyclonal knockout cells are suited for studying steroid metabolism disorders, mitochondrial dysfunction, and neurodevelopmental conditions. Applications include mitochondrial respiration assays (Seahorse), targeted metabolomics (LC-MS), validation via Western blotting and RT-qPCR, and immunofluorescence. The cells support screening of pharmacological chaperones and assessment of A??-induced toxicity, aiding drug discovery. For more information, please contact Ascent Research.
