The DNPH1 Knockout KYSE-30 Polyclonal Cells product is a pool of KYSE-30 esophageal squamous cell carcinoma cells with CRISPR/Cas9-mediated disruption of the DNPH1 gene. This polyclonal knockout cell population provides a loss-of-function model for studying nucleotide pool sanitization and genomic stability. The polyclonal format retains cellular heterogeneity, suitable for pooled screening and functional genomics experiments. The cells are prepared using a validated gene-editing approach to disrupt DNPH1 expression without clonal isolation.
The KYSE-30 cell line was established from a well-differentiated esophageal squamous cell carcinoma of a 64-year-old male patient. As a representative model of esophageal squamous cell carcinoma, KYSE-30 exhibits squamous differentiation features and relevant genomic alterations. The host cell background enables context-dependent investigation of how nucleotide metabolism and oxidative stress response contribute to esophageal cancer biology. Its robust growth characteristics and compatibility with standard assays ensure reproducible experimental setups.
DNPH1 encodes a nucleotide pool sanitizing enzyme that hydrolyzes 8-oxo-dGDP to 8-oxo-dGMP, preventing incorporation of oxidized guanine nucleotides into DNA and RNA. Its expression is transcriptionally activated by c-Myc, linking oncogenic proliferation to nucleotide pool fidelity. DNPH1 interacts with PCNA, NUDT15, and NUDT5, and functions upstream of base excision repair factors OGG1 and APE1 by limiting the substrate pool for their repair activities. This molecular network integrates oxidative stress signaling, nucleotide metabolism, and genome maintenance, with DNPH1 serving as a critical gatekeeper against oxidative DNA damage-induced mutagenesis.
In KYSE-30 esophageal cancer cells, DNPH1 knockout enables dissection of how nucleotide pool control influences cancer phenotypes under oxidative stress. Given frequent c-Myc dysregulation in esophageal squamous cell carcinoma, this model is valuable for studying the c-Myc?CDNPH1?Cgenome stability axis. The knockout is expected to increase 8-oxo-dGDP levels and oxidative DNA damage, revealing potential therapeutic vulnerabilities. This system also supports investigation of DNPH1??s roles in neurodevelopmental disorders and other oxidative damage-related malignancies.
The polyclonal knockout cells are suitable for applications including western blotting and RT-qPCR for DNPH1 expression analysis, 8-oxo-dG ELISA for oxidized nucleotide quantification, immunofluorescence and comet assays for DNA damage assessment, and flow cytometry for cell cycle profiling. RNA-seq under oxidative stress conditions can elucidate transcriptomic responses. These assays facilitate mechanistic studies and drug screening efforts targeting nucleotide metabolism and base excision repair in esophageal squamous cell carcinoma. For more information, contact Ascent Research.
