The HIBADH Knockout HAP1 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal population of HAP1 cells carrying a disrupted HIBADH gene locus. This reagent provides a loss-of-function model for the mitochondrial enzyme 3-hydroxyisobutyrate dehydrogenase, enabling investigation of valine degradation and downstream metabolic pathways. The polyclonal format ensures a heterogeneous pool of edited cells, reflecting variable knockout genotypes while maintaining robust population-level functional ablation of HIBADH activity.
HAP1 is a near-haploid human cell line derived from a male chronic myeloid leukemia patient, exhibiting an adherent fibroblast-like morphology. Its near-haploid karyotype simplifies genetic manipulation, as disruption of a single allele is sufficient to achieve functional knockout, making it an ideal host for high-throughput genetic screening and targeted gene knockout models. Widely used in functional genomics, HAP1 cells provide a clean background for studying metabolic and signaling pathways without the complexity of diploid gene redundancy.
HIBADH encodes a mitochondrial enzyme that catalyzes the NAD+-dependent oxidation of 3-hydroxyisobutyrate to methylmalonate semialdehyde, a key step in the valine catabolic pathway. This reaction links branched-chain amino acid degradation to the TCA cycle through propionyl-CoA and succinyl-CoA. The enzyme sits downstream of HIBCH and upstream of ALDH6A1, functioning within a pathway that includes BCKDHA, BCKDHB, and ACADSB. Its activity is regulated by PPARGC1A and mitochondrial biogenesis signals, and its disruption can lead to accumulation of 3-hydroxyisobutyrate, potentially altering NAD+/NADH ratios and TCA cycle flux.
In the HAP1 background, knockout of HIBADH provides a physiologically relevant model to dissect valine metabolism and its connection to mitochondrial energy production. The loss of HIBADH is expected to impair propionate metabolism and may mimic metabolic disturbances observed in 3-hydroxyisobutyric aciduria and methylmalonic acidemia. This cellular model allows researchers to study the consequences of blocked valine degradation, including altered levels of TCA cycle intermediates and potential metabolic acidosis phenotypes, in an isogenic context.
This product is suitable for a range of research applications, including valine metabolism studies, mitochondrial dysfunction research, and drug target validation. Experimentally, loss of HIBADH protein can be confirmed by Western blot, while transcript levels are assessable via RT-qPCR. Functional metabolic consequences can be evaluated through metabolite profiling of 3-hydroxyisobutyrate and TCA cycle intermediates, NAD+/NADH ratio measurements, and Seahorse-based metabolic flux analysis. It also serves as a control in CRISPR screens. For detailed technical specifications and availability, please contact Ascent Research.
