The DNER Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of gene-disrupted HAP1 cells, providing a loss-of-function model for the human DNER (Delta/Notch-like EGF-related receptor) gene. This product offers a heterogeneous pool of knockout alleles, enabling robust investigation of DNER-dependent signaling pathways without clonal selection bias. The polyclonal format is particularly suited for pooled screening approaches and studies requiring representation of diverse genetic perturbations within the Notch signaling axis.
The HAP1 cell line is a near-haploid human chronic myeloid leukemia (CML) cell line with fibroblast-like morphology, originally derived from the KBM-7 cell line. Its haploid karyotype simplifies genetic manipulation and facilitates the generation of clear loss-of-function phenotypes, making it an ideal host for CRISPR-based knockout studies. HAP1 cells are widely used in functional genomics, drug target discovery, and high-throughput screening due to their ease of culture and genetic tractability.
DNER encodes a type I transmembrane protein that functions as an activating ligand for Notch receptors, including Notch1 and Notch2. Upon ligand-receptor engagement, the Notch intracellular domain (NICD) is proteolytically released by the gamma-secretase complex, then translocates to the nucleus where it forms a transcriptional activation complex with RBPJ and MAML. This complex drives expression of downstream targets such as HES1, HEY1, MYC, and CCND1, thereby orchestrating cell fate decisions. DNER activity is regulated by proneural transcription factors including ASCL1 (MASH1) and neurogenins, and its signaling is modulated by Deltex E3 ubiquitin ligases. Cross-talk with other Notch ligands like JAG1 and DLL1 further refines the signaling output.
In the HAP1 haploid background, DNER disruption abrogates Notch activation specifically through this ligand, allowing dissection of DNER-dependent versus ligand-independent Notch signaling. The model is particularly relevant for studying neurogenesis and gliogenesis, given DNER’s established role in neural development. Moreover, as a CML-derived line, it enables exploration of DNER’s potential involvement in cancer cell signaling and oncogenic pathways. This knockout system thus serves as a valuable tool for both neurobiology and oncology research, linking Notch pathway components to disease-relevant phenotypes.
Researchers can employ these polyclonal knockout cells in a variety of assays to interrogate Notch pathway dynamics. Typical applications include Notch reporter luciferase assays to measure transcriptional activity, western blotting for NICD and HES1 protein levels, RT-qPCR analysis of Notch target gene expression, and immunofluorescence to assess DNER localization. Functional studies such as cell proliferation assays and co-culture Notch activation experiments are also feasible. These cells are well-suited for drug target validation, high-throughput chemical screening, and functional genomics investigations of the Notch signaling network. For further information or technical support, please contact Ascent Research.
