The IDH3G Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout population targeting the IDH3G gene in HAP1 cells. This heterogeneous pool enables loss-of-function studies without clonal selection, avoiding clone-specific artifacts. IDH3G encodes the regulatory gamma subunit of the mitochondrial isocitrate dehydrogenase complex, pivotal for TCA cycle flux and NADH generation. The polyclonal format offers a population-level model suitable for robust metabolic and signaling analyses.
Derived from KBM-7 chronic myeloid leukemia cells, HAP1 is an adherent near-haploid human cell line. Its haploidy reduces genetic redundancy, simplifying knockout interpretation. The immortalized hematopoietic background retains cancer-relevant metabolic features, and the adherent morphology supports diverse assay platforms, including live-cell imaging and metabolic flux analyzers.
IDH3G is an essential regulatory subunit of the NAD+-dependent mitochondrial IDH complex, interacting with IDH3A and IDH3B catalytic subunits to decarboxylate isocitrate into ??-ketoglutarate (??-KG) with concurrent NADH production. The complex is allosterically regulated by ADP/ATP ratios. Upstream, transcription factors MYC, HIF-1??, PGC-1??, and the deacetylase SIRT3 control IDH3G expression, linking TCA cycle activity to mitochondrial biogenesis and redox state. Consequently, IDH3G knockout impairs ??-KG and NADH production, reduces electron transport chain efficiency, and may perturb glutamate dehydrogenase-dependent anaplerosis, disrupting cellular redox homeostasis.
In HAP1 leukemia cells, IDH3G knockout models metabolic vulnerabilities of hematopoietic cancers, where TCA cycle rewiring and glutamine-driven ??-KG production support proliferation. The loss of the regulatory subunit creates a metabolic bottleneck, limiting NADH generation and potentially forcing reliance on alternative pathways like glutaminolysis. This system is ideal for probing metabolic reprogramming and identifying synthetic lethal targets in leukemia, with the haploid background ensuring clean phenotypic readouts.
These polyclonal knockout cells are suited for Seahorse metabolic flux assays (OCR/ECAR), NAD+/NADH ratio quantification, and ??-KG measurements by mass spectrometry. Validation can be performed by Western blot for IDH3 subunits and RT-qPCR for TCA cycle genes. Cell viability under metabolic stress (e.g., glucose deprivation) enables drug sensitivity screening. Applications extend to redox biology and mitochondrial dysfunction studies. For technical inquiries or custom options, contact Ascent Research.
