The HTRA1 Knockout MCF-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the MCF-7 human breast adenocarcinoma cell line. This product features CRISPR/Cas9-mediated disruption of the HTRA1 gene, resulting in a heterogeneous pool of cells with ablated HTRA1 expression. The polyclonal format provides a robust loss-of-function model while preserving potential genetic diversity, avoiding artifacts associated with clonal selection.
The MCF-7 cell line is a well-characterized epithelial cell model isolated from the pleural effusion of a 69-year-old Caucasian female with metastatic mammary adenocarcinoma. These cells express estrogen receptor and are widely used to study hormone-responsive breast cancer. Their luminal epithelial phenotype and well-defined signaling networks make them a preferred system for investigating tumor suppressor functions and oncogenic pathways.
HTRA1 encodes a secreted serine protease that targets key components of the extracellular matrix and the TGF-?? signaling axis. It is transcriptionally regulated by p53, DNA damage, oxidative stress, and E2F1, and functions as a tumor suppressor by cleaving the type II TGF-?? receptor, fibronectin, and decorin. This cleavage attenuates TGF-?? signaling, reducing phosphorylation of SMAD2 and SMAD3, which in turn diminishes transcription of pro-oncogenic genes. HTRA1 also interacts with TGF-??1 and insulin-like growth factor-binding proteins, integrating extracellular proteolysis with growth factor signaling. Disruption of HTRA1 releases this brake, potentially enhancing TGF-?? pathway activity and SMAD-dependent gene expression.
In MCF-7 cells, HTRA1 loss-of-function is particularly relevant for dissecting the tumor-suppressive roles of extracellular proteases in estrogen receptor-positive breast cancer. Since TGF-?? signaling can exert both tumor-suppressive and pro-metastatic effects depending on context, the HTRA1 knockout model allows precise investigation of how HTRA1-mediated cleavage of the TGF-?? receptor II and matrix substrates shifts signaling outcomes. This model is valuable for exploring crosstalk between p53-regulated apoptosis, extracellular matrix remodeling, and TGF-??-driven epithelial?Cmesenchymal transition.
Typical applications include western blotting for SMAD2/3 phosphorylation to assess TGF-?? pathway activation, TGF-?? luciferase reporter assays, migration and invasion assays, and extracellular matrix degradation assays to monitor proteolytic activity. Co-immunoprecipitation can be used to identify HTRA1 substrates, and proliferation assays help evaluate growth phenotypes. These cells are suitable for investigating tumor suppressor mechanisms, screening for anti-metastatic factors, and studying regulation of TGF-?? signaling. For further technical details or custom inquiries, please contact Ascent Research.
