The DNPH1 Knockout K-562 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population of K-562 cells harboring targeted disruption of the DNPH1 gene. This product is supplied as a heterogeneous pool of edited cells, offering researchers a versatile loss-of-function model to study DNPH1 in a leukemia background without the clonal biases inherent to single-cell-derived lines. The knockout was generated using ribonucleoprotein-mediated gene editing to introduce disruptive mutations, resulting in a polyclonal knockout model suitable for functional genomics and drug response studies.
The host K-562 cell line is a widely used suspension lymphoblastoid line originally established from the pleural effusion of a 53-year-old female with chronic myeloid leukemia (CML) in blast crisis. K-562 cells carry the BCR-ABL fusion gene characteristic of CML and serve as a classic model for hematopoietic differentiation and leukemia research. Their ability to undergo erythroid and megakaryocytic differentiation under appropriate stimuli, combined with their robust proliferative capacity, makes them an ideal platform for investigating oncogenic signaling and therapeutic vulnerabilities.
DNPH1 (2??-deoxynucleoside 5??-phosphate N-hydrolase 1) encodes an enzyme that catalyzes the N-glycosidic bond cleavage of 2??-deoxynucleoside 5??-phosphates, releasing free nucleobases and 2-deoxyribose 5-phosphate. This reaction is a rate-limiting step in the nucleotide salvage pathway, providing precursors for DNA synthesis and cell proliferation. DNPH1 functions as a homodimer transcriptionally activated by c-Myc. It supports dNTP generation channeled into DNA replication by DNA polymerase alpha, and intersects with deoxycytidine kinase (DCK) and thymidine kinase 1 (TK1). Thus, DNPH1 sits at a critical nexus linking c-Myc-driven transcriptional programs to nucleotide homeostasis.
In the K-562 CML context, where BCR-ABL signaling converges on c-Myc to sustain high proliferation rates, DNPH1 knockout is expected to severely compromise the salvage-dependent replenishment of dNTP pools. This disruption likely triggers replication stress, cell cycle arrest, and enhanced apoptosis, especially under conditions that challenge nucleotide biosynthesis. The model is therefore highly relevant for dissecting the metabolic dependencies of c-Myc-addicted leukemia cells and for evaluating the mechanistic basis of sensitivity to nucleoside-analog chemotherapeutics such as cytarabine and gemcitabine, which rely on salvage pathway enzymes for their activation.
Applications of the DNPH1 Knockout K-562 Polyclonal Cells include detailed investigation of the nucleotide salvage pathway’s role in leukemia cell proliferation and survival, assessment of chemosensitivity to antimetabolite drugs, and exploration of c-Myc oncogenic signaling networks. Researchers can perform cell proliferation assays (MTT, BrdU), apoptosis analyses (Annexin V/PI), nucleotide pool quantification by LC-MS, DNPH1 and c-Myc expression profiling via western blot and RT-qPCR, drug sensitivity dose-response curves, and flow cytometric cell cycle analysis. These polyclonal cells provide a robust platform for target validation and drug discovery efforts in hematological malignancies. For further information, please contact Ascent Research.
