GOT1 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which the gene encoding cytoplasmic aspartate aminotransferase (GOT1) has been disrupted in the human HAP1 cell line. This product is supplied as a heterogeneous pool of edited cells, enabling loss-of-function studies without the selective pressures associated with monoclonal expansion. The polyclonal format avoids the confounding effects of clonal variation while providing a robust model for investigating GOT1-dependent metabolic processes. Researchers can employ this knockout population for comparative analyses against wild-type HAP1 cells, utilizing standard assays to dissect the enzyme’s role in amino acid metabolism and redox homeostasis.
The HAP1 host cell line is a near-haploid human cell model derived from the KBM-7 chronic myeloid leukemia line. Its predominantly haploid karyotype simplifies functional genomics by allowing single-allele disruptions to produce clear phenotypes, reducing issues of genetic redundancy that complicate diploid systems. HAP1 cells express a range of metabolic and signaling pathways relevant to multiple tissue types, and their adherent growth properties and stable genetic background make them highly suitable for high-throughput screening, metabolomic profiling, and mitochondrial function analyses. This cell model has been widely adopted for CRISPR-based knockout studies, particularly for dissecting metabolic networks.
GOT1 encodes the cytoplasmic isoform of aspartate aminotransferase, which catalyzes the reversible transamination of aspartate and alpha-ketoglutarate into oxaloacetate and glutamate. This reaction is a critical component of the malate-aspartate shuttle, a key mechanism for transferring reducing equivalents across the mitochondrial membrane, linking glycolysis to oxidative phosphorylation. GOT1 is regulated upstream by transcription factors such as c-Myc, FOXO, NRF2, HIF1??, and glucocorticoids, and its activity influences levels of downstream metabolites including oxaloacetate, glutamate, aspartate, and malate. It functionally interacts with mitochondrial GOT2, malate dehydrogenases MDH1 and MDH2, glutamate dehydrogenase GLUD1, the mitochondrial aspartate-glutamate carrier SLC25A11, and the glutamate transporter SLC1A3. These molecular partners position GOT1 at a nexus of amino acid catabolism, gluconeogenesis, and the tricarboxylic acid cycle.
In the near-haploid HAP1 context, disruption of GOT1 provides a clean genetic background to evaluate metabolic dependencies. Loss of GOT1 activity can impair the malate-aspartate shuttle, potentially affecting NADH oxidation, maintenance of cytosolic redox state, and anaplerotic flux. This knockout model is particularly valuable for studying metabolic reprogramming in cancer, where GOT1 is often upregulated to support glutamine metabolism and nucleotide biosynthesis. By comparing polyclonal knockout and wild-type cells, researchers can assess compensatory mechanisms??such as upregulation of GOT2 or alternative transaminases??and define the essentiality of this shuttle in various nutrient environments. HAP1’s inherent genetic tractability further permits double knockout experiments or rescue studies with mutant GOT1 variants.
Typical applications of GOT1 Knockout HAP1 Polyclonal Cells extend across multiple fields, including cancer metabolism (glutamine addiction, redox balance), liver biology (gluconeogenesis, ammonia detoxification), and neurological disease models where aspartate aminotransferase deficiency is implicated. These cells are compatible with diverse experimental workflows: western blotting for protein expression analysis, spectrophotometric enzyme activity assays, RT-qPCR for transcript quantification, untargeted or targeted metabolomics via mass spectrometry, mitochondrial function assays (Seahorse), and stable isotope tracing to map metabolic flux. The combination of a well-defined haploid host and polyclonal knockout strategy offers a flexible and reproducible tool for probing GOT1 function. For further details, please contact Ascent Research.
