The DNPH1 Knockout 143B Polyclonal Cells are a CRISPR/Cas9-edited population of human 143B osteosarcoma cells bearing targeted gene disruption of DNPH1. Composed of a heterogeneous pool of edited cells, this product provides a consistent loss-of-function model while avoiding clonal selection artifacts. CRISPR/Cas9-mediated editing introduces site-specific genomic alterations that abrogate functional DNPH1 protein production across the polyclonal population. This format facilitates robust depletion of DNPH1 activity, enabling reproducible phenotypic analysis.
The 143B cell line originates from a human osteosarcoma and is a widely used model in cancer research. These cells exhibit rapid proliferation and transformed properties characteristic of osteosarcoma, making them suitable for investigating tumor cell biology, oncogenic signaling, and therapeutic responses. The osteosarcoma background provides a pathologically relevant environment for studying nucleotide metabolism regulators like DNPH1 in the context of bone cancer.
DNPH1 functions as a 2′-deoxynucleoside 5′-phosphate N-hydrolase that hydrolyzes deoxynucleoside monophosphates (dNMPs), thereby regulating intracellular nucleotide pools. Its expression is transcriptionally activated by the c-Myc oncoprotein, placing it downstream of a major mitogenic and metabolic facilitator. By limiting dNMP accumulation, DNPH1 helps prevent misincorporation errors during DNA synthesis and protects against DNA damage-induced apoptosis. The enzyme is thus integrated into the DNA damage response network, interacting with dNMP substrates, DNA polymerases, and repair enzymes to maintain genomic fidelity.
In 143B osteosarcoma cells, DNPH1 knockout provides a means to dissect the c-Myc?CDNPH1?Cnucleotide metabolism axis. c-Myc overexpression drives metabolic reprogramming in many cancers, and DNPH1 may buffer the resultant nucleotide pool imbalances, potentially contributing to chemoresistance. Loss of DNPH1 in this model can reveal vulnerabilities in nucleotide homeostasis and DNA repair pathways that are exploitable for therapeutic intervention. This knockout background thus allows exploration of how c-Myc-driven nucleotide metabolism impacts osteosarcoma cell survival and drug sensitivity.
These polyclonal knockout cells are suitable for a range of applications, including RT-qPCR and western blotting for target validation, LC-MS-based nucleotide pool quantification, and cell viability assays (MTT, CellTiter-Glo) to assess proliferation. DNA damage responses can be evaluated via COMET assay or ??-H2AX staining, and apoptosis measured by Annexin V or caspase activity tests. The model supports drug target validation, study of c-Myc downstream functions, and synthetic lethality screening. For further details, please contact Ascent Research.
