The DLAT Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for functional studies of the dihydrolipoamide S-acetyltransferase (DLAT) gene in a near-haploid chronic myeloid leukemia background. This polyclonal knockout model enables loss-of-function analysis without clonal isolation, providing a heterogeneous pool of edited cells ideal for population-level assays. The product is generated by CRISPR/Cas9-mediated disruption of the DLAT locus in HAP1 cells, creating a versatile tool for investigating metabolic regulation and signaling networks.
HAP1 is a near-haploid human cell line derived from a male patient with chronic myelogenous leukemia. Its near-haploid karyotype simplifies genetic manipulation and ensures high editing efficiency, making it a robust platform for CRISPR-based gene disruption. As a leukemia cell line, HAP1 retains cancer-relevant metabolic and signaling features, including dependence on glycolysis and altered mitochondrial function, which are particularly relevant for dissecting metabolic pathways.
DLAT encodes the E2 subunit of the pyruvate dehydrogenase (PDH) complex, catalyzing acetyl group transfer from pyruvate to coenzyme A, generating acetyl-CoA. This reaction bridges glycolysis and the tricarboxylic acid (TCA) cycle, tightly regulated by upstream kinases PDK1?C4 (phosphorylating and inhibiting E1??) and phosphatases PDP1?C2 (dephosphorylating and activating E1??). DLAT requires lipoic acid as a cofactor and interacts with PDHA1/PDHB (E1), DLD (E3), and PDHX (E3BP) to form the functional PDH complex. Acetyl-CoA fuels the TCA cycle and ATP production, and serves as a substrate for SIRT3-mediated histone acetylation, linking metabolism to epigenetic regulation. DLAT disruption ablates PDH activity, forcing metabolic rerouting and reprogramming.
In the HAP1 chronic myeloid leukemia background, DLAT knockout generates a powerful model for examining how loss of PDH complex function impacts cancer cell metabolism. HAP1 cells, with their near-haploid genome, provide a clean genetic background that minimizes confounding variables. Disruption of DLAT is expected to uncouple glycolysis from the TCA cycle, mimicking conditions seen in pyruvate dehydrogenase deficiency and certain cancers where PDH activity is suppressed. This model allows researchers to study compensatory metabolic pathways, such as glutamine-dependent reductive carboxylation, and to interrogate the role of DLAT in sustaining proliferation and survival under nutrient stress.
The DLAT Knockout HAP1 Polyclonal Cells suit cancer metabolism research, metabolic disorder modeling, and epigenetic regulation studies. Typical assays include PDH activity measurements, Seahorse metabolic flux analysis, acetyl-CoA quantification, and metabolomics profiling. Western blotting and RT-qPCR confirm DLAT disruption and downstream effects, while proliferation and apoptosis assays evaluate functional consequences. This polyclonal knockout population is valuable for drug sensitivity studies targeting metabolic vulnerabilities. For further information, contact Ascent Research.
