The DNMBP Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population engineered to disrupt the human DNMBP gene in HEK293T cells. This heterogeneous cell pool provides a versatile loss-of-function model for investigating DNMBP-dependent processes without clonal selection biases. The gene editing approach leads to target-gene disruption, enabling researchers to study the absence of DNMBP function in a physiologically relevant epithelial context.
HEK293T is a human embryonic kidney epithelial cell line that stably expresses the SV40 large T-antigen, renowned for its high transfection efficiency and robust growth. These adherent, semi-attached cells are a workhorse for heterologous protein expression and lentivirus production. Their epithelial origin also renders them suitable for examining cytoskeletal dynamics, receptor trafficking, and cell motility. The DNMBP knockout cells retain these advantageous properties while allowing precise dissection of DNMBP-mediated pathways.
DNMBP functions as a Cdc42-specific guanine nucleotide exchange factor (GEF) and scaffold protein, acting downstream of integrin signaling and growth factor receptors such as EGFR and PDGFR. It directly interacts with dynamin and N-WASP, coupling Cdc42 activation to actin polymerization through the Arp2/3 complex. This coordination is essential for receptor-mediated endocytosis, where DNMBP-promoted actin assembly drives vesicle scission, and for cell migration, where it regulates focal adhesion dynamics and lamellipodia protrusion. Disruption of DNMBP thus impairs key cytoskeletal and trafficking processes controlled by the Cdc42 signaling axis.
In HEK293T cells, DNMBP knockout abolishes Cdc42-specific GEF activity, providing a powerful system to study endocytic trafficking defects and migration abnormalities. The epithelial nature of these cells makes them particularly relevant for exploring the role of DNMBP in proteinuric kidney diseases such as steroid-resistant nephrotic syndrome and in cancer cell invasion. This polyclonal knockout population enables investigation of disease mechanisms and the validation of therapeutic targets within a genetically controlled yet heterogeneous cellular environment.
Typical applications include Western blotting and RT-qPCR for confirming gene disruption, immunofluorescence to assess actin architecture and focal adhesions, and transferrin uptake assays to quantify receptor-mediated endocytosis. Researchers can also perform wound healing migration assays and co-immunoprecipitation experiments to evaluate DNMBP interactions with dynamin, N-WASP, and Cdc42. Additionally, the cells are suitable for drug screening campaigns targeting GTPase modulators or trafficking pathways. For further details or to request a quotation, please contact Ascent Research.
