The DNPH1 Knockout MCF-7 Polyclonal Cells constitute a CRISPR/Cas9-mediated gene-disrupted cell population derived from the MCF-7 human breast cancer cell line. This polyclonal knockout model establishes loss-of-function of DNPH1, a key enzyme in nucleotide metabolism, enabling dissection of nucleotide salvage pathway dependencies in a well-characterized breast cancer background. The heterogeneous knockout pool captures diverse editing outcomes, providing a robust platform for studying DNPH1’s impact without clonal selection artifacts.
MCF-7 is a metastatic mammary adenocarcinoma cell line originally established from a pleural effusion of a patient with breast cancer. These cells retain estrogen receptor positivity and wild-type TP53 status, making them a widely used model for hormone-responsive breast cancer research. Their adherent epithelial morphology and well-documented growth characteristics facilitate standardized assays in cancer biology, drug response, and signal transduction studies.
DNPH1 (2??-deoxynucleoside 5??-monophosphate N-glycosidase) catalyzes the hydrolysis of dNMPs into free nucleobases and deoxyribose phosphate, directly modulating cellular dNTP pools and DNA synthesis. DNPH1 is transcriptionally regulated by the c-Myc oncoprotein, placing it within the c-Myc target gene network that promotes nucleotide biosynthesis. Its activity intersects with key nucleotide metabolic enzymes including thymidine kinase 1 (TK1), deoxycytidine kinase (dCK), and downstream nucleotide kinases, thereby influencing pyrimidine and purine metabolism pathways essential for genomic integrity and cell cycle progression.
Disruption of DNPH1 in the estrogen receptor-positive MCF-7 context provides a valuable model to investigate how nucleotide salvage dysregulation alters breast cancer cell fitness. Since c-Myc-driven metabolic reprogramming is frequently amplified in aggressive tumors, this knockout model helps elucidate the dependency of hormone-responsive breast cancer cells on DNPH1-mediated nucleotide recycling. The system is particularly relevant for probing synthetic lethal interactions and compensatory mechanisms that maintain dNTP homeostasis under replication stress conditions commonly encountered in tumor biology.
Researchers can apply this polyclonal knockout population in a spectrum of functional assays, including cell viability assessments (MTT/resazurin), colony formation, and flow cytometric cell cycle analysis to evaluate proliferation defects. Complementary metabolomic profiling and LC-MS-based dNTP quantification enable precise measurement of nucleotide pool alterations, while RT-qPCR and Western blotting confirm gene disruption and downstream expression changes. Nucleotide salvage activity assays further characterize substrate-level metabolic flux. These tools facilitate studies in drug resistance mechanisms and synthetic lethality screens targeting nucleotide metabolism. For further technical specifications or ordering information, please contact Ascent Research.
