The ARHGAP5 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from HT29 human colorectal adenocarcinoma cells, containing heterogeneous ARHGAP5 gene disruptions. This pool enables loss-of-function studies without clonal selection, providing a relevant model for investigating p190-B RhoGAP function in cancer biology.
HT29 cells, a well-characterized colon adenocarcinoma line with epithelial morphology, serve as a standard intestinal epithelial model. They form polarized monolayers, retain key signaling pathways, and are widely used in colorectal cancer research, offering a physiologically appropriate background to study ARHGAP5-mediated processes.
ARHGAP5 encodes p190-B, a GTPase-activating protein that inactivates RhoA, Rac1, and Cdc42 by accelerating GTP hydrolysis. It is regulated by EGF, PDGF, integrin engagement, SRC family kinases, and PKC??. Active Rho GTPases signal through ROCK1/2, PAK1, LIMK, and cofilin to control actin cytoskeleton reorganization, focal adhesion dynamics, and cell migration. p190-B interacts with RhoA-GTP, Rac1-GTP, Cdc42-GTP, p120-catenin, FAK, SRC, and paxillin, positioning it at key adhesive and signaling hubs.
ARHGAP5 knockout in HT29 cells leads to hyperactivation of RhoA, Rac1, and Cdc42, promoting stress fiber formation, increased focal adhesion turnover, and enhanced cell migration and invasion. This mimics aggressive colorectal carcinoma behavior, making the model valuable for studying cytoskeletal-driven metastasis and testing Rho pathway interventions.
Applications include Rho GTPase activation assays, Western blotting for phospho-MLC and phospho-cofilin, immunofluorescence of actin and focal adhesions, wound healing, transwell invasion, and cell adhesion assays. The cells support inhibitor screening and RT-qPCR analysis of downstream targets. Contact Ascent Research for further information.
