The DNER Knockout K-562 Polyclonal Cells represent a genetically heterogeneous pool of K-562 cells engineered via CRISPR/Cas9 to disrupt the DNER gene. This polyclonal knockout population enables loss-of-function studies of DNER-dependent Notch signaling without the need for single-cell cloning. The product is well-suited for population-level assays and provides a versatile starting point for biochemical, pharmacological, and differentiation experiments.
The parental human K-562 cell line was derived from the bone marrow of a 53-year-old female with chronic myeloid leukemia in blast crisis. K-562 cells carry the Philadelphia chromosome, resulting in the BCR-ABL oncogenic fusion, and are extensively utilized as a model system for hematopoietic stem cell biology, erythroid differentiation, and leukemogenesis. Their robust proliferation and multi-lineage differentiation capacity make them an ideal host for dissecting signaling pathways in a malignant hematopoietic environment.
DNER (Delta/Notch-like EGF-related receptor) is a type I transmembrane protein that functions as a contact-dependent Notch ligand. Upon binding to NOTCH1 or NOTCH2, DNER induces ADAM10-mediated S2 cleavage followed by ??-secretase-dependent release of the Notch intracellular domain (NICD). Nuclear NICD complexes with the transcription factor CSL/RBPJ and the coactivator MAML to activate transcription of target genes such as HES1, HES5, HEY1, p21, and CCND1. DNER expression is controlled by neural bHLH transcription factors (NEUROD1, ASCL1) and promoter methylation, and the protein plays a key role in cell fate specification, differentiation, and proliferation, with emerging evidence of tumor-suppressive functions.
Introducing DNER knockout into K-562 cells creates a unique platform to examine Notch ligand function within a BCR-ABL-driven leukemia model. Loss of DNER may perturb Notch-mediated regulation of apoptosis, cell cycle progression, and drug sensitivity, potentially influencing the differentiation potential of K-562 along erythroid or megakaryocytic lineages. This polyclonal model allows direct assessment of DNER??s contribution to Notch activation in hematopoiesis, circumventing compensation by other ligands.
These knockout cells are applicable to a broad range of techniques, including RT-qPCR for quantifying Notch target gene changes, western blotting for NICD detection, co-immunoprecipitation to confirm disrupted DNER-NOTCH1 binding, and flow cytometry for surface DNER profiling. Functional assays such as proliferation, differentiation, and luciferase reporter assays for Notch activity can be performed. The product is well-suited for drug discovery screens targeting the Notch pathway, toxicology evaluations, and studies of BCR-ABL?CNotch crosstalk in leukemia. For additional technical details, please contact Ascent Research.
