The DNPH1 knockout KYSE-150 polyclonal cells from Ascent Research are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human esophageal squamous cell carcinoma cell line KYSE-150. This product features targeted disruption of the DNPH1 gene using CRISPR/Cas9-mediated genome editing, generating a heterogeneous pool of cells with DNPH1 loss of function. The polyclonal format avoids clonal selection artifacts and is well-suited for population-level analyses of nucleotide metabolism and cancer biology.
KYSE-150 is a widely used esophageal squamous cell carcinoma model established from a poorly differentiated tumor of a 49-year-old Japanese female. Retaining key characteristics of esophageal cancer, this cell line provides a relevant context for studying oncogenic signaling, metabolic reprogramming, and therapeutic vulnerabilities. Esophageal squamous cell carcinoma remains a challenging malignancy, underscoring the need for robust model systems to evaluate novel targets such as nucleotide salvage enzymes.
DNPH1 (2′-deoxynucleoside 5′-monophosphate N-glycosidase) catalyzes the hydrolysis of dNMPs to deoxyribose 5-phosphate and nucleobases, a critical step in nucleotide salvage that regulates dNTP pools required for DNA synthesis and repair. The MYC transcription factor transcriptionally activates DNPH1, promoting nucleotide recycling in proliferating cells. DNPH1 thus influences dNTP homeostasis, DNA synthesis, and cell proliferation. It interacts with nucleotide metabolism enzymes and potentially DNA repair factors, linking nucleotide metabolism to genome stability.
In KYSE-150 cells, DNPH1 disruption enables dissection of nucleotide salvage in supporting the metabolic demands of esophageal cancer. MYC amplification, common in this cancer type, may drive DNPH1 expression, making the MYC-DNPH1 axis a candidate vulnerability. Loss of DNPH1 is expected to impair dNTP balance and DNA damage responses, potentially sensitizing cells to genotoxic stress or nucleotide analogs. This polyclonal knockout model allows assessment of population-level effects on growth, metabolism, and drug sensitivity.
Applications include investigating nucleotide metabolism and DNA damage response in esophageal cancer, and screening for inhibitors of nucleotide salvage. Knockout validation can be performed by Western blot and RT-qPCR, while functional studies may employ LC-MS for nucleotide pool analysis, MTS or BrdU proliferation assays, and ??H2AX immunofluorescence for DNA damage. Drug sensitivity testing with nucleotide analogs further explores therapeutic potentials. For additional information, contact Ascent Research.
