The ELANE Knockout T-47D Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the T-47D human breast cancer cell line. This product enables loss-of-function studies of the ELANE gene, which encodes neutrophil elastase, a serine protease implicated in extracellular matrix degradation and inflammatory processes. The polyclonal knockout format provides a heterogeneous population with targeted gene disruption, suitable for experiments where clonal homogeneity is not required and bulk population-level phenotypic analysis is desired.
The T-47D host cell line originates from a pleural effusion of an invasive ductal carcinoma and is widely employed as a model of estrogen receptor-positive (ER+) and progesterone receptor-positive (PR+) breast cancer. These cells retain features of luminal epithelial differentiation and are extensively characterized for hormone-responsive signaling pathways, making them a relevant backdrop for studying the interplay between hormonal regulation and tumor microenvironment remodeling. Their well-established use in endocrine therapy resistance studies further expands the utility of this knockout model.
Neutrophil elastase, encoded by ELANE, is a serine protease that cleaves elastin and other extracellular matrix (ECM) components such as collagen IV and fibronectin. Its activity is tightly regulated by endogenous inhibitors including SERPINA1 (??1-antitrypsin), SERPINB1, and secretory leukocyte protease inhibitor (SLPI). Upstream, ELANE expression is induced by granulocyte colony-stimulating factor (G-CSF/CSF3), transcription factors CEBPE and GFI1, and pro-inflammatory cytokines like tumor necrosis factor ?? (TNF??) and interleukin-1?? (IL-1??). Downstream, elastase activity promotes the release of IL-8 and activation of matrix metalloproteinases (MMPs), notably MMP9, thereby contributing to ECM remodeling, inflammatory cell recruitment, and a microenvironment conducive to invasion.
In the T-47D breast cancer context, ectopic ELANE activity has been correlated with enhanced invasive potential due to proteolysis of basement membrane components and activation of pro-invasive signaling cascades. Disruption of ELANE in this ER+/PR+ model provides a clean system to dissect the cell-autonomous contribution of serine protease activity to hormone-dependent and -independent tumor cell migration, decoupled from neutrophil-derived elastase. This knockout population is particularly useful for investigating whether ELANE-mediated ECM degradation cooperates with or bypasses classical hormone receptor signaling in driving metastatic behavior.
This polyclonal knockout cell product is suited for a wide range of experimental workflows including protease activity assays using fluorogenic substrates, Boyden chamber migration and invasion assays, gelatin zymography for MMP profiling, Western blotting for ECM protein analysis, immunofluorescence staining of matrix components, and transcriptomic analyses (RNA-seq) to map signaling network perturbations. Additionally, it supports screening of elastase inhibitors and evaluating therapeutic candidates targeting tumor microenvironment proteolysis. For technical inquiries and ordering, please contact Ascent Research.
