The ADAM10 Knockout 769-P Polyclonal Cells are a ready-to-use CRISPR/Cas9-edited polyclonal knockout population of human 769-P renal cell carcinoma cells engineered for targeted disruption of the ADAM10 gene. ADAM10 encodes a transmembrane metalloproteinase that functions as a master sheddase, cleaving ectodomains of diverse substrates. This polyclonal format provides a heterogenous gene-disrupted cell pool, minimizing clonal bias and better reflecting the genetic diversity observed in tumor environments, while enabling immediate use in loss-of-function studies.
The 769-P cell line is a well-characterized clear cell renal cell carcinoma epithelial model, retaining oncogenic features and widely used in kidney cancer research. It offers a clinically relevant context for investigating ADAM10-mediated processes in carcinoma, as it endogenously expresses relevant substrates and signaling molecules.
ADAM10 is a transmembrane sheddase that cleaves ectodomains of Notch1, E-cadherin, and APP. It is regulated by inhibitors TIMP-1 and TIMP-3 and activated by PKC, calcium, and EGF. In the Notch pathway, ligand engagement triggers ADAM10-mediated S2 cleavage, followed by gamma-secretase S3 cleavage to release NICD, which translocates to the nucleus and transcriptionally activates targets such as HES1 via RBP-J. ADAM10 also sheds E-cadherin to produce soluble E-cadherin, disrupting adherens junctions and promoting cell motility, and cleaves APP to generate sAPP??. The sheddase interacts with tetraspanins Tspan12, Tspan15, and Tspan33, which guide its substrate specificity, and cooperates with the gamma-secretase complex. Additional substrates include HB-EGF, CD44, and CX3CL1, linking ADAM10 to EGFR transactivation, cell adhesion, and chemokine signaling.
Disruption of ADAM10 in the 769-P renal carcinoma background provides a powerful model to interrogate the role of sheddase activity in cancer cell behavior. Since clear cell RCC commonly exhibits aberrant Notch pathway activation and altered E-cadherin expression, ADAM10 loss-of-function allows dissection of its contributions to tumor cell proliferation, migration, invasion, and epithelial-to-mesenchymal transition. The polyclonal knockout population avoids clonal selection artifacts and preserves some genetic heterogeneity, more closely approximating the genetic landscape of tumors, and is suitable for studying the collective response to ADAM10 disruption in a pooled setting.
Researchers can utilize these cells in a wide array of functional assays, including Notch reporter assays to measure pathway activity, sheddase activity assays using APP or HB-EGF shedding, Western blotting and RT-qPCR for ADAM10 and downstream targets, transwell migration and invasion assays, cell adhesion assays, wound healing, and drug sensitivity screens with ADAM10 inhibitors. This model also supports epithelial-mesenchymal transition studies and investigation of ADAM10??s role in renal cancer metastasis. For additional technical details, protocols, or ordering information, please contact Ascent Research.
