The ARHGAP17 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for the targeted disruption of the ARHGAP17 gene. This heterogeneous pool of HT29-derived cells enables loss-of-function studies without the biases introduced by single-cell cloning. The polyclonal format provides a representative cross-section of genetic modifications, preserving biological variability while abrogating ARHGAP17 expression.
The HT29 line, isolated from a 44-year-old female colorectal adenocarcinoma, is an established model of intestinal epithelium. These cells form polarized monolayers, express tight junction proteins (occludin, ZO-1), and differentiate into enterocyte-like cells, making them ideal for studying epithelial polarity and colorectal cancer.
ARHGAP17 encodes a Rho GTPase-activating protein that preferentially inactivates the small GTPases Cdc42 and Rac1 by accelerating their GTP hydrolysis. In epithelial cells, ARHGAP17 is recruited to tight junctions via interactions with angiomotin (AMOT) and angiomotin-like proteins (AMOTL1/L2), where it locally suppresses Cdc42 and Rac1 activity. This regulation modulates downstream effectors including PAK kinases and WASP/N-WASP, thereby controlling actin cytoskeleton remodeling and the stabilization of tight junction complexes containing occludin and ZO-1. ARHGAP17 function is influenced by upstream signals such as the CDX2 transcription factor, TGF-?? pathway activation, and pro-inflammatory cytokines like TNF-??. Disruption of ARHGAP17 disrupts the balance of Rho GTPase signaling, leading to altered cell polarity, compromised epithelial barrier integrity, and enhanced migratory potential.
In HT29 cells, ARHGAP17 knockout weakens tight junctions, reduces TEER, and enhances migration and invasion. This phenocopies aspects of colorectal cancer progression, where ARHGAP17 is often downregulated, and its loss may synergize with inflammatory signals such as TNF-??. The model allows dissection of how ARHGAP17 maintains epithelial polarity via Amot and Patj interactions, and how its absence facilitates epithelial-mesenchymal transition-like changes.
These polyclonal knockout cells are suitable for profiling tight junction protein expression by western blotting and immunofluorescence (occludin, ZO-1), functional barrier assays (TEER), and cell motility studies (scratch wound healing, transwell migration/invasion). They also enable biochemical analysis of Rho GTPase activity (Cdc42/Rac1 pulldown), co-immunoprecipitation of ARHGAP17 binding partners, and qPCR-based gene expression analysis. Drug screening for barrier restoration and assessment of tumor suppressors in colon cancer are additional applications. For detailed technical specifications, contact Ascent Research.
