The ITGB1 Knockout HT29 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma line. This heterogeneous pool harbors disruption of the ITGB1 gene, which encodes the integrin ??1 subunit, offering a versatile loss-of-function system free from monoclonal artifacts. The cells are delivered as a ready-to-use population suitable for immediate culture and downstream applications.
HT29 cells originate from a primary colon carcinoma of a 44-year-old female and represent a well-established model of the intestinal epithelial barrier. These mucin-producing epithelial cells display adherent growth and express markers characteristic of colorectal adenocarcinoma, including E-cadherin and cytokeratins. Their tumorigenic background makes them particularly relevant for investigating integrin-dependent processes in colorectal cancer biology.
Integrin ??1 serves as the essential ?? subunit for heterodimeric receptors that bind extracellular matrix proteins such as fibronectin, collagen, and laminin. Ligand engagement recruits intracellular adaptors talin, vinculin, and paxillin, which activate focal adhesion kinase (FAK). FAK signaling propagates through Src to PI3K/Akt and MAPK/ERK cascades, while also regulating Rho GTPases Rac1 and Cdc42. Upstream factors including TGF-??, EGF, and the SP1 transcription factor modulate ITGB1 expression, with talin and kindlin governing integrin activation. These pathways collectively influence migration, proliferation, and survival via downstream effectors like cyclin D1 and MMP-9.
Disruption of ITGB1 in HT29 cells ablates ??1 integrin surface expression, causing severe deficiencies in adhesion to fibronectin and collagen. This loss attenuates FAK and MAPK/ERK phosphorylation, diminishing cell motility and proliferative capacity. The model thereby uncouples integrin signaling from oncogenic circuits such as PI3K/Akt and Wnt, offering a powerful tool to dissect contributions to tumor cell invasiveness, ECM remodeling, and epithelial-mesenchymal transition. Unlike clonal lines, this polyclonal population preserves biological variability, enhancing physiological relevance.
Applications include adhesion assays on ECM substrates, transwell migration/invasion studies, and western blotting for phospho-FAK (Tyr397) and phospho-Akt (Ser473). Immunofluorescence of vinculin and paxillin illuminates focal adhesion dynamics, while RNA-seq and flow cytometry for integrin levels enable comprehensive characterization. Drug sensitivity tests with integrin antagonists facilitate preclinical evaluation, and the model proves valuable for research into colorectal cancer progression, integrin inhibitor pharmacology, chemoresistance, and metastasis. For additional information, please contact Ascent Research.
