The ADAM10 Knouckout DLD-1 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population derived from the DLD-1 human colorectal adenocarcinoma cell line. This model disrupts the ADAM10 gene, encoding a transmembrane metalloprotease responsible for ectodomain shedding of numerous cell surface proteins. As a polyclonal pool, the population comprises cells carrying heterogeneous targeted disruptions at the ADAM10 locus, enabling robust loss-of-function studies without clonal selection bias. The knockout is suitable for investigating ADAM10-dependent signaling and substrate processing in a genomically defined background.
DLD-1 is a widely characterized epithelial cell line isolated from a male patient with colorectal adenocarcinoma. These cells retain key features of colorectal tumors, including mutations in APC, KRAS, and TP53, and are commonly employed to model colorectal cancer progression, cell adhesion, and intestinal epithelial barrier dysfunction. Their adherent epithelial morphology and rapid proliferation support reproducible culture conditions. The line is a standard choice for dissecting oncogenic signaling cascades and evaluating therapeutic targets in vitro.
ADAM10 functions as a principal sheddase, mediating regulated intramembrane proteolysis of substrates such as Notch1, E-cadherin, amyloid precursor protein (APP), and EGFR ligands. Proteolytic activation by Furin and calcium influx, modulated by TIMP1 and GPCR signaling, allows ADAM10 to cleave target ectodomains. It interacts with tetraspanins Tspan12, Tspan15, and CD9, and coordinates with the gamma-secretase complex to release intracellular fragments (e.g., Notch intracellular domain, NICD). Downstream, NICD translocates to the nucleus, associating with RBP-J to drive Hes1 transcription, while EGFR ligand shedding activates MAPK signaling. These pathways converge on cell fate decisions, adhesion, and proliferation.
In DLD-1 cells, ADAM10 disruption abolishes ligand-dependent Notch1 cleavage and downstream signaling, attenuates E-cadherin ectodomain shedding, and reduces EGFR ligand release. Consequently, this knockout model likely impairs Notch-driven transcriptional programs, weakens cell?Ccell adhesion, and dampens EGFR signaling. Given the basal pathway activity in DLD-1, the cells provide a valuable system to delineate ADAM10-specific contributions to colorectal oncogenesis, particularly in regulating intestinal epithelial differentiation, stemness, and sensitivity to targeted agents. The polyclonal format preserves population heterogeneity, making it suitable for studying stochastic signaling adaptation.
Researchers can utilize this polyclonal knockout model for functional dissection of Notch and EGFR signaling pathways, identification of novel ADAM10 shedding substrates, and assessment of drug resistance mechanisms in colorectal cancer. Compatible assays include quantitative Western blotting and RT-qPCR for pathway component expression, flow cytometry for surface protein analysis, immunofluorescence to visualize junctional integrity, and functional tests such as colony formation and cell adhesion assays. The model also supports intestinal epithelial barrier studies under inflammatory conditions. For additional information or technical inquiries, please contact Ascent Research.
