The IMPDH2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 cell line, providing a heterogeneous loss-of-function model for studying IMPDH2. In this product, CRISPR/Cas9 technology disrupts the IMPDH2 gene across a pool of cells, yielding a mixed population with varied mutations. This polyclonal format avoids single-cell cloning bottlenecks and facilitates direct assessment of gene disruption effects on cellular physiology.
HAP1 is a human near-haploid cell line derived from KBM-7 chronic myeloid leukemia cells, exhibiting adherent fibroblast-like morphology. Its largely haploid karyotype (except for disomic chromosome 8) enables recessive phenotypes from single-gene disruptions, making it ideal for knockout studies. HAP1 cells are widely used for CRISPR-based functional genomics, pathway analysis, and high-throughput screens due to their robust growth and genetic tractability.
IMPDH2 catalyzes the rate-limiting conversion of inosine monophosphate (IMP) to xanthosine monophosphate (XMP) in de novo guanine nucleotide biosynthesis, a critical step for generating GTP essential for DNA synthesis and cell proliferation. IMPDH2 expression is transcriptionally activated by oncogenic factors MYC and E2F, while growth factor signaling through mTOR upregulates translation to meet increased nucleotide demand. The enzyme cooperates with IMPDH1 and forms filaments that undergo allosteric inhibition by GTP, providing feedback control. Disruption of IMPDH2 therefore depletes intracellular GTP pools, attenuates DNA replication, and reduces proliferation, as salvage pathway compensation is insufficient.
In the IMPDH2 Knockout HAP1 Polyclonal Cells, loss of de novo guanine synthesis creates a potent system for probing purine metabolism dependency, with the near-haploid background intensifying metabolic phenotypes. This model holds particular relevance for cancer biology, as many tumors upregulate IMPDH2 to sustain rapid proliferation, and for immunosuppression research, since IMPDH2 is the molecular target of mycophenolic acid. By comparing knockout and wild-type populations, researchers can dissect nucleotide deprivation effects on cell cycle progression, apoptosis, and metabolic reprogramming, identifying vulnerabilities exploitable by therapeutic interventions.
Typical research uses include cancer cell proliferation studies, drug target validation, and mechanistic investigations of immunosuppression. Standard characterization employs Western blotting to confirm IMPDH2 protein loss, LC-MS-based nucleotide profiling to monitor IMP and GTP changes, MTT or similar viability assays, and flow cytometry for cell cycle analysis. Metabolomics further reveals systems-level alterations in purine metabolism. For additional details or customer support, please contact Ascent Research.
