The AMDHD2 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the AMDHD2 gene in the HAP1 cell line. This product provides a loss-of-function model for investigating the role of AMDHD2 in histidine catabolism and associated metabolic pathways. The polyclonal knockout cells are derived from a heterogeneous pool, enabling robust population-level analyses without the bottleneck effects of clonal selection. Researchers can utilize these cells to study the functional consequences of AMDHD2 gene disruption on amino acid metabolism and cancer cell physiology.
The HAP1 host cell line is a human near-haploid chronic myeloid leukemia cell model derived from the KBM-7 line. Characterized by its adherent growth and near-haploid karyotype, HAP1 cells are a widely adopted system for genetic knockout studies due to the ease of generating complete gene disruptions. The leukemic origin of HAP1 cells makes them particularly relevant for exploring metabolic reprogramming in hematopoietic malignancies, providing a physiologically appropriate context for studying AMDHD2-dependent processes in cancer.
AMDHD2 encodes formiminoglutamase, which catalyzes the final step of histidine degradation by hydrolyzing N-formimino-L-glutamate into L-glutamate and formamide. This reaction is crucial for maintaining nitrogen balance and supplying glutamate for the TCA cycle. The activity of AMDHD2 is integrated into a broader metabolic network, with upstream regulators including mTORC1 signaling and the MYC transcription factor, both of which modulate amino acid metabolism. AMDHD2 functionally interacts with the histidine catabolic enzymes histidine ammonia-lyase (HAL) and urocanate hydratase (UROC1), as well as the upstream paralog AMDHD1. Disruption of AMDHD2 results in reduced glutamate production and potential accumulation of the intermediate N-formimino-L-glutamate, which can be monitored by targeted metabolomics.
In the HAP1 leukemic background, loss of AMDHD2 may compromise the cell??s ability to utilize histidine as a carbon and nitrogen source, potentially creating a metabolic vulnerability. Given that cancer cells often exhibit altered amino acid metabolism to support rapid proliferation, this knockout model enables the dissection of AMDHD2??s contribution to anaplerotic reactions and redox homeostasis. The near-haploid genome simplifies genetic analysis, allowing researchers to correlate AMDHD2 deficiency with specific phenotypic outcomes, such as altered sensitivity to chemotherapeutics that target one-carbon metabolism or glutaminolysis. Thus, these cells are a valuable tool for understanding metabolic dependencies in hematological cancers.
Typical research applications include metabolic flux analysis using LC-MS to measure N-formimino-L-glutamate (FIGLU) levels and histidine consumption, as well as functional assays such as cell proliferation, colony formation, and drug sensitivity screening. The cells are suitable for downstream molecular characterization by Western blotting and RT-qPCR to assess pathway alterations. By integrating these approaches, researchers can investigate the role of AMDHD2 in cancer metabolic reprogramming and evaluate its potential as a therapeutic target. For more information, please contact Ascent Research.
