The HTRA1 Knockout T-47D Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the HTRA1 gene in the T-47D human breast ductal carcinoma cell line. This loss-of-function model enables the investigation of HTRA1’s biological roles and signaling functions in a controlled in vitro setting. The polyclonal population, generated via CRISPR/Cas9-mediated gene disruption, provides a versatile tool for studying the consequences of HTRA1 depletion without the constraints of single-cell clonal variation.
The host T-47D cell line was originally derived from the pleural effusion of a 54-year-old female with infiltrating ductal carcinoma. These cells are estrogen receptor (ER)-positive, progesterone receptor (PR)-positive, and belong to the luminal A molecular subtype, characterized by TP53 mutation and estrogen-dependent growth. Their hormone-responsive nature and well-defined genetic background make them a widely employed model for breast cancer research, particularly for studying endocrine therapy responses and hormone-driven tumor progression.
HTRA1 encodes a secreted serine protease that negatively regulates TGF-beta and BMP signaling by cleaving ligands such as TGF-beta and BMP4, and receptors like TGFBR2, thereby reducing SMAD2/3 and SMAD1/5/8 phosphorylation. It also degrades ECM components including fibronectin and decorin. Upstream regulators include p53, RB1, TGF-beta, and HIF1A, while downstream effects modulate apoptosis, migration, and SMAD-mediated transcription.
In the context of T-47D cells, HTRA1 knockout is particularly significant due to the cell line’s luminal A breast cancer origin and TP53 mutant status. Loss of HTRA1’s tumor-suppressive protease activity is expected to enhance TGF-beta signaling, promoting epithelial-mesenchymal transition (EMT), cell migration, and aberrant ECM dynamics??processes that contribute to breast cancer progression. Moreover, the interplay between HTRA1 and p53-dependent pathways in this estrogen-responsive background offers a unique platform to dissect cooperative tumor suppressor mechanisms and hormonal influences on protease-mediated signaling.
This polyclonal knockout population supports applications such as Western blotting for SMAD phosphorylation, RT-qPCR for TGF-beta target genes, and luciferase reporter assays. Functional assays include proliferation, migration, invasion, apoptosis analysis by flow cytometry, and ECM degradation. The model is useful for drug response studies targeting TGF-beta signaling or tumor suppressor restoration. For additional technical specifications and ordering details, please contact Ascent Research.
