The HTRA1 Knockout HT29 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal cell population derived from HT29 colorectal adenocarcinoma cells, engineered for disruption of the HTRA1 gene. This heterogeneous knockout pool contains a mixture of cells with targeted gene disruptions introduced via CRISPR/Cas9, providing a loss-of-function model that avoids clonal artifacts and captures a spectrum of editing outcomes.
The HT29 parental line was established from a primary colorectal adenocarcinoma of a 44-year-old female patient diagnosed with Dukes?? stage B colorectal cancer. These adherent epithelial cells retain key features of the intestinal epithelium and are widely used in colorectal cancer research for studying oncogenic signaling, drug resistance, and tumor cell biology.
HTRA1 is a secreted serine protease with established tumor-suppressive functions. It cleaves extracellular matrix (ECM) substrates including fibronectin and decorin, and directly cleaves TGF-?? receptor II, thereby downregulating TGF-??/SMAD signaling. HTRA1 also inhibits Wnt pathway activity by promoting ??-catenin degradation. Upstream regulators such as TGF-??1, BMP4, and p53 control HTRA1 expression, while its downstream effects extend to casein, amyloid precursor protein, and insulin-like growth factor binding proteins. HTRA1 interacts with tubulin and PDZ-domain proteins, linking its protease activity to cytoskeletal and scaffolding networks. Through these mechanisms, HTRA1 constrains proliferation, migration, and invasion.
In HT29 cells, which harbor mutations in APC and TP53, HTRA1 knockout is expected to further perturb TGF-?? and Wnt signaling, amplify ECM remodeling, and enhance malignant properties. This polyclonal model thus provides a relevant genetic context to dissect how loss of HTRA1 cooperates with existing oncogenic lesions to drive colorectal cancer progression and metastasis, offering insights into the tumor-suppressive role of HTRA1 within a human adenocarcinoma background.
Researchers can employ these cells in a variety of assays, including western blotting and RT-qPCR for HTRA1 loss verification, MTT or BrdU proliferation assays, transwell migration/invasion assays, and TGF-??-responsive luciferase reporter assays. Additional applications encompass substrate cleavage assays to monitor protease activity, immunofluorescence for ECM organization, and apoptosis assays. These cells are suitable for tumor suppression studies, ECM dynamics analysis in cancer, signaling pathway interrogation, and disease modeling of age-related macular degeneration and osteoarthritis. For further information, contact Ascent Research.
