The DNER Knockout HT29 Polyclonal Cells provide a ready-to-use CRISPR/Cas9-edited polyclonal knockout population derived from the HT29 human colorectal adenocarcinoma cell line. In this model, targeted disruption of the DNER gene abolishes expression of the transmembrane Delta/Notch-like epidermal growth factor-related receptor, a key Notch ligand. As a heterogeneous knockout population, these cells enable robust loss-of-function studies of DNER-dependent signaling in an epithelial colorectal cancer background, eliminating the need for transient knockdown or pharmacological inhibition.
The HT29 cell line, originally established from a primary colorectal adenocarcinoma of a 44-year-old female, is a widely utilized model in cancer biology and preclinical drug evaluation. These adherent epithelial cells exhibit a differentiated colorectal phenotype under conventional culture conditions and retain intact core signaling networks, including the Notch pathway. Their well-documented genomic profile and reproducible growth characteristics make HT29 cells an ideal host for CRISPR/Cas9-mediated gene disruption aimed at pathway dissection.
DNER encodes a single-pass transmembrane protein that engages Notch receptors (NOTCH1, NOTCH2, NOTCH3) in a cell-contact-dependent manner. Upon ligand binding, the Notch extracellular domain is cleaved by ADAM10/TACE, followed by intramembrane proteolysis by the ??-secretase complex, generating the Notch intracellular domain (NICD). NICD translocates to the nucleus, where it forms a transcriptional activation complex with the DNA-binding protein RBPJ and the coactivator MAML1. This complex directly promotes expression of downstream targets, including the transcriptional repressors HES1, HEY1, and HEY2, as well as MYC, CCND1, and CDKN1A. Consequently, DNER knockout in HT29 cells interrupts this signaling cascade, leading to reduced transcription of Notch-responsive genes and altered cellular outcomes such as proliferation, differentiation, and survival.
In colorectal cancer, Notch signaling exerts context-dependent roles, influencing tumor initiation, stem cell self-renewal, differentiation, and drug resistance. By eliminating DNER-mediated Notch activation, this polyclonal HT29 knockout model enables researchers to discriminate ligand-specific contributions to these processes. The population-level knockout approach preserves heterogeneity akin to tumor cell populations, allowing assessment of signaling dynamics and responses to microenvironmental cues without clonal selection bias. This makes the model particularly valuable for studying Notch-driven oncogenic mechanisms and therapeutic vulnerabilities.
These polyclonal knockout cells are suitable for a broad array of experimental techniques, including Western blot and RT-qPCR quantification of DNER and Notch targets (e.g., HES1, HEY1), flow cytometry to monitor Notch receptor surface expression, and functional assays such as MTT proliferation, soft agar colony formation, and Transwell migration/invasion. They also support Notch-responsive luciferase reporter assays, co-immunoprecipitation of DNER-Notch complexes, transcriptome profiling by RNA-seq, and apoptosis detection via Annexin V staining. Applications include drug screening for Notch pathway inhibitors, functional genomics of cell fate determination, and cancer stem cell research. For additional product details or technical inquiries, please contact Ascent Research.
