The DMTN knockout 786-O polyclonal cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the DMTN gene, which encodes dematin, an actin-bundling protein. This loss-of-function model enables systematic investigation of dematin??s role in cytoskeletal organization and membrane stability within a cancer-relevant context. The polyclonal format captures a spectrum of editing outcomes, offering a representative phenotypic profile without clonal selection biases.
The host line, 786-O, is a well-characterized adherent human renal epithelial cell line derived from clear cell renal cell carcinoma (ccRCC). Its VHL deficiency leads to constitutive HIF stabilization, driving tumorigenic attributes such as angiogenesis and metabolic adaptation. This background is highly relevant for renal cancer research, including metastasis, drug resistance, and hypoxia-driven signaling.
Dematin serves as a key organizer of the cortical actin-spectrin network, directly interacting with spectrin (SPTAN1, SPTB), adducin (ADD1), actin (ACTB), and EPB41. It is regulated by upstream kinases PKA, PKC, and SRC, as well as calmodulin and mechanical stress. Downstream, dematin facilitates F-actin bundling, spectrin tetramerization, and anchoring of GLUT1 to the plasma membrane. DMTN ablation perturbs the RhoA-ROCK-LIMK-cofilin axis, increasing actin dynamics and cell motility, while also compromising cell adhesion complexes.
In 786-O cells, DMTN deletion recapitulates the reduced dematin expression observed in metastatic renal cell carcinoma, resulting in enhanced migration and invasion. The loss disrupts cortical actin integrity and spectrin-based membrane scaffolding, weakening adhesion and promoting a more aggressive phenotype. This model is particularly suited for dissecting the interplay between VHL-associated hypoxia pathways and actin cytoskeletal remodeling during epithelial-to-mesenchymal transition.
Typical experimental uses include transwell migration, scratch wound, and Matrigel invasion assays to quantify metastatic behavior. Immunofluorescence with phalloidin staining and co-immunoprecipitation enable visualization and biochemical validation of actin-spectrin network alterations. Western blotting, flow cytometry for GLUT1 surface localization, and apoptosis/proliferation assays further delineate functional consequences. The cells also support drug screening for anti-metastatic compounds and mechanistic studies of RhoA-driven cytoskeletal signaling. For detailed technical specifications or to order, please contact Ascent Research.
