The IQGAP3 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population engineered to disrupt the IQGAP3 gene (IQ motif containing GTPase activating protein 3) in the HEK293T human embryonic kidney cell line. This loss-of-function model provides a powerful tool for investigating the scaffold protein??s roles in cytoskeletal dynamics and cell signaling. The polyclonal nature offers a heterogeneous knockout background, suitable for population-level studies without clonal selection artifacts.
HEK293T cells are a widely utilized derivative of the HEK293 line, stably expressing the SV40 large T antigen to enable episomal replication of plasmids containing the SV40 origin. Their epithelial origin and high transfection efficiency make them a standard platform for viral production, protein expression, and functional genomics. The robust growth characteristics and well-characterized signaling networks of HEK293T cells provide a consistent cellular context for dissecting IQGAP3-dependent pathways.
IQGAP3 functions as a scaffold protein that integrates signals upstream of the actin cytoskeleton and cell cycle machinery. It directly interacts with and activates the small GTPases Rac1 and Cdc42, thereby promoting actin polymerization, lamellipodia formation, and directed cell migration. IQGAP3 also collaborates with APC and CLIP-170 to regulate mitotic spindle orientation, and potentiates Wnt/??-catenin signaling by stabilizing the ??-catenin destruction complex component APC, leading to increased cyclin D1 transcription. Upstream, IQGAP3 expression is induced by EGF and Wnt pathways, and is under negative regulation by miR-145. In turn, IQGAP3 scaffolds the PAK kinase downstream of Rac1/Cdc42, reinforcing cytoskeletal reorganization and cell cycle progression.
In HEK293T cells, IQGAP3 knockout allows dissection of its contributions to epithelial cell behavior, independent of tissue-specific oncogenic drivers. The polyclonal population provides a more physiologically diverse model compared to single clones, and is ideal for bulk assays such as immunoblotting and qPCR to assess pathway activation. Since HEK293T cells express relevant signaling receptors, this model enables analysis of EGF- and Wnt-induced cytoskeletal and proliferative responses, and facilitates co-immunoprecipitation studies of IQGAP3 complex formation with endogenous Rac1, Cdc42, and ??-catenin.
This knockout model is suited for functional studies of cancer cell migration and invasion via Transwell or wound-healing assays, cell cycle analysis by flow cytometry, and signaling network profiling through phospho-protein analysis. It supports drug target validation by assessing the impact of IQGAP3 loss on compound efficacy in proliferation or motility readouts. The cells can be combined with rescue experiments to map functional domains or used in pooled CRISPR screens for genetic interaction mapping. For further technical information, please contact Ascent Research.
