The ARHGAP11A Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the ARHGAP11A gene in human HEK293T cells. Using CRISPR/Cas9-mediated gene disruption, this model generates a heterogeneous pool of knockout cells, enabling robust loss-of-function studies of the Rho GTPase-activating protein ARHGAP11A in a widely used epithelial background. The polyclonal format captures diverse editing events, providing a versatile system for functional genomics and signaling research.
HEK293T cells are derived from human embryonic kidney epithelia, immortalized with adenovirus 5 DNA, and stably express the SV40 large T antigen. This cell line is extensively utilized for protein overexpression, lentivirus production, and cell signaling studies due to its high transfectability and well-characterized molecular framework. Its epithelial origin and reliable growth make it a suitable host for interrogating Rho GTPase-mediated cytoskeletal regulation.
ARHGAP11A functions as a GTPase-activating protein that stimulates GTP hydrolysis on Rho family GTPases, including RhoA, Rac1, and Cdc42, thereby inactivating them. This activity negatively regulates actin stress fiber formation, focal adhesion dynamics, and downstream transcriptional outputs. Upstream regulators include transcription factors MYC and E2F, and signaling by EGF and HGF via integrin adhesion. Downstream, ARHGAP11A suppresses ROCK-LIMK-cofilin signaling, myosin light chain phosphorylation, and FAK/paxillin clustering, ultimately limiting YAP/TAZ nuclear translocation and cyclin D1 expression. It directly interacts with GTP-bound RhoA, Rac1, and Cdc42, and opposing ARHGEFs restore GTP loading. This positions ARHGAP11A at a critical junction between growth factor inputs, cytoskeletal mechanics, and Hippo pathway control.
In HEK293T cells, ARHGAP11A knockout is predicted to increase active RhoA, Rac1, and Cdc42, leading to enhanced stress fiber assembly, focal adhesion maturation, and altered mechanotransduction. Elevated RhoA-GTP promotes actomyosin contractility and YAP/TAZ activity, which may drive proliferation and gene expression changes. The polyclonal nature of the knockout population avoids clonal selection bias, enabling population-level analyses of these interconnected processes in an epithelial context.
This knockout model is applicable to diverse experimental approaches: Rhotekin pull-down assays for RhoA-GTP quantification, immunofluorescence staining of F-actin and paxillin, RT-qPCR profiling of YAP target genes, and migration/invasion or proliferation assays. It is also suitable for high-throughput compound screening targeting Rho pathway components and co-immunoprecipitation studies with active Rho GTPases. For additional product information or custom requests, please contact Ascent Research.
