The DNPH1 Knockout T-47D Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the DNPH1 gene. This product provides a heterogeneous pool of T-47D cells carrying targeted disruptions in DNPH1, enabling researchers to interrogate the functional consequences of abrogated DNPH1 expression without clonal selection. The knockout model serves as a valuable tool for dissecting nucleotide metabolism, DNA repair, and oncogenic signaling in a well-characterized breast cancer background.
The parental T-47D cell line is a human breast ductal carcinoma epithelial line derived from a pleural effusion of a 54-year-old female with infiltrating ductal carcinoma. These cells are estrogen receptor-positive (ER+), progesterone receptor-positive (PR+), and androgen receptor-positive (AR+), and belong to the luminal A molecular subtype. T-47D cells are extensively utilized as a model for hormone-dependent breast cancer, exhibiting growth responses to estrogens and other steroid hormones, and they recapitulate key features of ER+ breast tumor biology, including hormone-driven proliferation and tumorigenesis.
DNPH1 encodes a nucleoside monophosphate phosphohydrolase that hydrolyzes deoxynucleoside monophosphates (dNMPs) to deoxynucleosides and inorganic phosphate, a reaction central to nucleotide salvage and dNTP pool homeostasis. The enzyme is transcriptionally regulated by c-Myc and exhibits cell cycle-dependent expression, linking its activity to proliferation. DNPH1 functions within a network that includes nucleotide kinases (dCK, TK1, TK2, dGK), phosphoribosyltransferases (APRT, HPRT), and nucleoside transporters. By controlling dNMP levels, DNPH1 modulates DNA synthesis fidelity and repair efficiency, impacting genomic stability. This interplay engages ATM/ATR damage response and p53 pathways, placing DNPH1 at the intersection of metabolism and genome maintenance.
In the context of T-47D ER+ breast cancer cells, DNPH1 knockout disrupts the delicate balance of nucleotide pools, potentially impairing both DNA replication and repair capacity. This perturbation is particularly relevant given the hormone-sensitive nature of the host line, as c-Myc is a known downstream effector of estrogen signaling and frequently overexpressed in luminal breast cancers. The loss of DNPH1 may attenuate proliferation rates and render cells more susceptible to genotoxic agents such as gemcitabine, which targets nucleotide metabolism. Thus, this polyclonal knockout model enables dissection of the c-Myc?CDNPH1 axis and its contribution to hormone-dependent growth, offering insights into metabolic vulnerabilities that could be exploited therapeutically.
Recommended applications encompass a broad spectrum of experimental modalities. Researchers can employ these cells to investigate nucleotide salvage pathway dynamics in breast cancer, validate c-Myc target gene functions, and probe DNA damage response mechanisms. Representative assays include RT-qPCR and Western blotting for expression analysis, LC-MS-based quantification of dNTP pools, cell proliferation and clonogenic survival assays, ??H2AX immunofluorescence and comet assays for DNA damage assessment, and flow cytometry for cell cycle profiling. Drug sensitivity screens, particularly with nucleoside analogs, and migration/invasion assays further expand the utility. For more information on product specifications and technical support, please contact Ascent Research.
