The DNPH1 Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-mediated gene disruption model targeting the DNPH1 gene in the human SK-HEP-1 cell line. This polyclonal knockout cell population provides a loss-of-function system for investigating DNPH1-dependent processes. The product is supplied as a pool of edited cells, enabling robust and reproducible assays without the selection artifacts associated with single-cell clones. By abrogating DNPH1 expression, researchers can dissect its role in nucleotide pool sanitation and DNA damage responses directly in a liver sinusoidal endothelial context.
The SK-HEP-1 host cell line was originally derived from the ascitic fluid of a patient with liver adenocarcinoma and exhibits endothelial-like characteristics. These cells have become a widely accepted model for liver sinusoidal endothelial cells, as they form a functional barrier, regulate hepatic stellate cell activation, and mediate endocytosis and immune surveillance. Their unique phenotype makes SK-HEP-1 cells particularly valuable for studying liver-specific vascular biology and pathophysiology. The endothelial-like properties of SK-HEP-1 cells are maintained in this knockout population, allowing physiologically relevant experiments.
DNPH1 encodes a dNTPase that hydrolyzes oxidized nucleotides, most notably 8-oxo-dGTP, to prevent their mutagenic incorporation into DNA. The enzyme is transcriptionally upregulated by the c-Myc oncogene, linking nucleotide pool maintenance to proliferative signaling. DNPH1 functions downstream of c-Myc and reduces 8-oxo-dGTP pools, thereby protecting DNA polymerases from incorporating 8-oxoguanine into nascent DNA. It interacts with nucleotide metabolism enzymes and potentially with DNA polymerases, forming a critical node in the DNA damage response and nucleotide metabolism pathways. Disruption of DNPH1 leads to accumulation of 8-oxo-dGTP and increased genomic instability.
In the context of liver sinusoidal endothelial cells, loss of DNPH1 is expected to heighten sensitivity to oxidative stress and replicate the mutagenic environment characteristic of c-Myc-driven genomic instability. The SK-HEP-1 knockout model thus serves as a platform to explore the intersection of nucleotide metabolism and endothelial cell biology, with relevance to liver cancer and genomic instability syndromes. It enables the study of how endothelial cells cope with oxidative DNA damage, a common feature of the hepatic microenvironment.
Researchers can employ these polyclonal knockout cells in a variety of applications, including the assessment of DNA damage response kinetics, evaluation of mutagenesis mechanisms, and investigation of nucleotide metabolism in liver endothelial cells. The model is particularly suited for drug sensitivity testing of oxidative stress modulators and for exploring c-Myc-dependent pathways. Representative assays include Western blotting, RT-qPCR, comet assay, 8-oxo-dGTP ELISA, immunofluorescence for 8-oxoguanine, RNA-seq, cell proliferation assays, and drug sensitivity assays. For further information or to discuss custom applications, please contact Ascent Research.
