The ARHGEF17 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colon epithelial adenocarcinoma cell line. This product provides a loss-of-function model for ARHGEF17, which encodes a RhoA-specific guanine nucleotide exchange factor (GEF). The polyclonal pool, obtained by CRISPR/Cas9-mediated gene disruption, retains biological heterogeneity and is well suited for population-level functional studies without clonal bottleneck effects.
The HT29 cell line was originally established from a primary colorectal adenocarcinoma of a 44-year-old Caucasian female and is extensively utilized as an in vitro model for colorectal cancer research. These cells display characteristic epithelial morphology and tumorigenic properties in xenograft assays. The line carries well-documented mutations in the tumor suppressor genes APC and TP53 and exhibits a microsatellite stable (MSS) phenotype, which recapitulates the genomic features of a large subset of sporadic colorectal carcinomas.
ARHGEF17 functions as a dedicated guanine nucleotide exchange factor that activates RhoA by catalyzing the exchange of GDP for GTP, leading to accumulation of RhoA-GTP. Active RhoA engages downstream effectors such as ROCK1 and mDia1, which in turn regulate LIM kinase (LIMK), cofilin phosphorylation, and myosin light chain activity to drive actin polymerization, stress fiber formation, and cellular contractility. Upstream signals from G protein-coupled receptors, integrin-mediated adhesion, and mechanical tension modulate ARHGEF17 function. Additionally, ARHGEF17 interacts with the tight junction scaffold protein PALS1 (MPP5) and influences signaling through serum response factor (SRF) and focal adhesion kinase (FAK), thereby linking extracellular cues to cytoskeletal reorganization and adhesive structures.
In HT29 colorectal adenocarcinoma cells, disruption of ARHGEF17 is predicted to impair RhoA-dependent cell adhesion and migration. Loss of ARHGEF17 likely compromises tight junction integrity and actin cytoskeletal dynamics, rendering the knockout population a valuable tool for dissecting how RhoA signaling contributes to epithelial barrier function and metastatic behavior in a genetic background deficient in APC and TP53.
Researchers can apply this polyclonal knockout model in diverse experimental workflows. Biochemical assays include western blotting for RhoA-GTP and phosphorylated myosin light chain, and RhoA activation assays (G-LISA). Immunofluorescence staining of F-actin and vinculin enables visualization of cytoskeletal alterations. Functional studies encompass scratch wound migration, transwell invasion, and transepithelial electrical resistance (TEER) measurements to evaluate barrier function. The model also supports chemosensitivity testing and confocal microscopy of tight junction proteins, as well as cell proliferation analyses to probe tumor-suppressive roles of ARHGEF17. For further information, please contact Ascent Research.
