This product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the UM-UC-3 human bladder carcinoma cell line, with targeted disruption of the CASP6 gene. The polyclonal pool comprises cells harboring diverse loss-of-function mutations, providing a robust model for studying CASP6 function while avoiding clonal selection artifacts. This knockout system is suited for investigating the roles of executioner caspase-6 in apoptosis, inflammation, and neurodegeneration within a clinically relevant urothelial carcinoma background.
The UM-UC-3 line originates from a male patient with transitional cell carcinoma (urothelial carcinoma) and is a well-established model of invasive bladder cancer. This adherent cell line exhibits aggressive features, including high migratory capacity, survival signaling, and resistance to anoikis. UM-UC-3 cells are routinely applied to study tumor progression, metastasis, and drug resistance, making them an appropriate host for evaluating the functional consequences of CASP6 knockout.
CASP6 encodes a downstream executioner caspase activated by initiator caspases (CASP8 via death receptor ligands FasL/TNF-alpha; CASP9 via cytochrome c/Apaf-1) and granzyme B. Active CASP6 cleaves lamin A/C, cytokeratin 18, PARP1, SATB1, huntingtin, tau, beta-catenin, and alpha-tubulin to drive apoptotic nuclear and cytoskeletal dismantling. Regulatory interactions include XIAP, c-IAP1, c-FLIP, Bcl-2 proteins, and HSP70. Non-apoptotic roles encompass inflammation and tau cleavage in neurodegeneration.
In bladder cancer, caspase-6 contributes to apoptosis susceptibility and invasive potential. Aberrant expression may promote chemoresistance. The UM-UC-3 CASP6 knockout model allows investigation of how loss of caspase-6 alters sensitivity to agents like cisplatin and doxorubicin, modifies migration/invasion, and affects lamin A/C fragmentation. This system is ideal for nuclear morphology studies and for distinguishing caspase-6-dependent non-apoptotic functions in tumor cell motility and matrix remodeling.
Applications include flow cytometry with Annexin V/PI staining, TUNEL assays, and Western blotting for cleaved substrates (lamin A, PARP1) and initiator caspase activation. Fluorogenic or colorimetric substrates measure caspase-6 activity. RT-qPCR confirms CASP6 ablation; immunofluorescence visualizes lamin A disruption. Migration/invasion assays (Boyden chamber, wound healing) evaluate metastatic behavior. For further information, please contact Ascent Research.
