The IDH3B Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of HAP1 cells with disruption of the IDH3B gene, encoding the ?? subunit of mitochondrial NAD-dependent isocitrate dehydrogenase (IDH3). This loss-of-function model is intended for research on TCA cycle regulation, mitochondrial redox homeostasis, and metabolic diseases. The polyclonal knockout context avoids single-clone bias while maintaining a near-haploid genetic background, enabling metabolic flux analysis, disease modeling, and genetic screening.
HAP1 is a near-haploid human cell line derived from KBM-7 chronic myeloid leukemia, haploid except for chromosome 8. This genomic simplicity allows direct genotype-phenotype correlations without heterozygous compensation. Its hematopoietic origin suits blood metabolism and malignancy studies, and its robust growth facilitates CRISPR editing, making HAP1 a key model for functional genomics and high-throughput screens.
IDH3B encodes the ?? subunit of the IDH3 complex, which irreversibly converts isocitrate to ??-ketoglutarate (??-KG) while reducing NAD+ to NADH in the TCA cycle. IDH3 activity is allosterically activated by ADP and inhibited by NADH, and is modulated by SIRT3 deacetylation. Its transcription is driven by NRF1 and TFAM. The ?? subunit interacts with IDH3A and IDH3G to form the functional holoenzyme. ??-KG downstream enters glutamate/glutamine metabolism and is oxidized by ??-ketoglutarate dehydrogenase. Loss of IDH3B disrupts NADH production, TCA flux, and ??-KG-dependent pathways, critically affecting cellular energetics and redox balance.
In the haploid HAP1 background, IDH3B knockout generates a clean loss-of-function model without compensatory alleles. Disruption reduces mitochondrial NADH output, alters ??-KG levels, and shifts carbon flux from glutamate/glutamine, ideal for studying metabolic reprogramming in cancer and mitochondrial disease. The near-haploid state enables synthetic lethality screens and chemical-genetic profiling to uncover vulnerabilities linked to IDH3B deficiency, relevant to retinitis pigmentosa and other mitochondrial disorders.
These cells support metabolic flux analysis, NADH/NAD+ ratio measurements, mitochondrial respiration assays (Seahorse), and cell viability tests under stress. They enable mechanistic studies of IDH3 in mitochondrial disorders and retinitis pigmentosa via RT-qPCR and western blotting. Haploid genetic screens can identify synthetic lethal partners or TCA cycle regulators, advancing cancer metabolism research and drug discovery. For technical inquiries, contact Ascent Research.
