The CASP6 Knockout T-47D Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout population derived from the T-47D human breast cancer cell line, with targeted disruption of the CASP6 gene encoding the executioner caspase-6. This polyclonal product preserves the natural genetic diversity of the edited pool, providing a robust loss-of-function model for investigating caspase-6-dependent processes without the biases of clonal selection.
T-47D cells were originally isolated from a pleural effusion metastasis of a breast ductal carcinoma and display an epithelial phenotype characteristic of hormone-responsive, estrogen receptor-positive (ER+) and progesterone receptor-positive (PR+) luminal A breast cancer. They maintain key signaling pathways governing hormone-dependent growth and apoptosis, making them a relevant host line for studying caspase-6 function in endocrine-sensitive oncology.
Caspase-6 is an effector cysteine protease activated downstream of initiator caspases caspase-8 and caspase-9, following intrinsic apoptosis (via APAF1/cytochrome c-mediated caspase-9 activation) or extrinsic apoptosis (triggered by death receptors such as Fas and TRAIL), and also by granzyme B. The protease cleaves critical substrates including lamin A/C, PARP1, keratin 18, alpha-tubulin, beta-catenin, SATB1, and huntingtin, driving nuclear lamina disassembly, cytoskeletal collapse, and inhibition of DNA repair. Its activity is modulated by interaction partners XIAP, caspase-3, heat shock protein Hsp70, and 14-3-3 proteins, forming a tightly regulated network central to apoptosis execution and implicated in axonal degeneration and neurodegenerative disorders.
In T-47D luminal breast cancer cells, CASP6 knockout impairs apoptotic execution and can alter cellular responses to chemotherapeutics and hormonal agents. This model enables dissection of caspase-6 contributions to drug-induced cell death and exploration of compensatory pathways that may emerge, offering insights into mechanisms of endocrine resistance and apoptosis dysregulation in breast cancer.
Researchers can employ these polyclonal knockout cells in a variety of assays, including apoptosis detection via Annexin V staining, caspase activity measurements, Western blot analysis of cleaved lamin A and PARP1, drug sensitivity profiling with MTT or CellTiter-Glo assays, immunofluorescence for nuclear morphology, and RT-qPCR for apoptotic gene expression. The product is also suitable for investigating neurodegenerative proteolytic events, screening caspase-6 inhibitors, and studying crosstalk between intrinsic and extrinsic apoptotic pathways. For further technical inquiries or ordering, please contact Ascent Research.
