The DNPH1 Knockout 786-O Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal population of human 786-O renal carcinoma cells carrying a targeted disruption of the DNPH1 gene. This polyclonal knockout cell pool offers a genetically heterogeneous loss-of-function model suitable for studying gene function in a population context, without clonal selection. The edited cell population is provided as a ready-to-use reagent for examining DNPH1-dependent processes in cancer cell biology.
The parental 786-O cell line is a widely established human clear cell renal cell carcinoma (ccRCC) model derived from a primary adenocarcinoma. 786-O cells harbor a naturally occurring inactivating mutation in the von Hippel-Lindau (VHL) tumor suppressor gene, leading to stabilization and constitutive activation of hypoxia-inducible factors HIF-1?? and HIF-2?? under normoxic conditions. This genetic background recapitulates a hallmark of ccRCC pathology and drives a pseudohypoxic transcriptional program that promotes tumor growth, angiogenesis, and metabolic reprogramming.
DNPH1 encodes a deoxynucleoside 5??-monophosphate hydrolase that catalyzes the hydrolysis of deoxynucleoside monophosphates (dNMPs) into deoxynucleosides and inorganic phosphate, thereby regulating the balance of nucleotide pools required for high-fidelity DNA replication and repair. DNPH1 expression is transcriptionally regulated by the c-Myc oncoprotein, linking oncogenic signaling directly to nucleotide metabolism. Within the cellular signaling network, DNPH1 acts downstream of c-Myc and upstream of DNA polymerases, interacting with enzymes of the nucleotide salvage pathway and factors involved in the DNA damage response. This positions DNPH1 as a mediator of c-Myc-driven metabolic adaptations that sustain genomic integrity in rapidly proliferating cancer cells.
In the 786-O ccRCC background, constitutive HIF activation often coexists with enhanced c-Myc activity, creating a metabolic context that relies on robust nucleotide supply. Disruption of DNPH1 function in these cells is expected to disturb deoxynucleotide homeostasis, potentially leading to insufficient dNTP pools for DNA synthesis and repair. This may result in replication stress, accumulation of DNA damage, and heightened vulnerability to genotoxic agents. Consequently, the DNPH1 knockout 786-O polyclonal cells serve as a valuable tool for dissecting the intersection of oncogenic signaling, nucleotide metabolism, and DNA damage responses in renal cell carcinoma.
Researchers can employ these knockout cells in a variety of experimental setups, including quantitative analysis of nucleotide pools, assessment of DNA damage markers such as ??-H2AX by immunofluorescence, cell proliferation and colony formation assays, and drug sensitivity screens with DNA-damaging chemotherapeutics like gemcitabine. Transcriptomic studies via RNA-seq can reveal the broader impact of DNPH1 loss on gene expression programs. The model is particularly suited for synthetic lethality screens and investigations into c-Myc-driven metabolic vulnerabilities in ccRCC. For additional information or technical inquiries, please contact Ascent Research.
