EGFR Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated by disruption of the epidermal growth factor receptor (EGFR) gene in the HEK293T human embryonic kidney cell line. This loss-of-function model enables the study of EGFR-dependent signaling without the influence of residual receptor activity, providing a powerful tool for investigating cellular processes regulated by this receptor tyrosine kinase.
HEK293T cells are a widely used derivative of the HEK293 line, transformed with adenovirus 5 DNA and stably expressing the SV40 large T antigen. This modification permits episomal replication of plasmids containing the SV40 origin of replication, resulting in high-level transient protein expression and robust viral packaging. Their epithelial morphology, rapid growth rate, and high transfection efficiency make HEK293T cells an ideal host for gene-editing applications, enabling scalable production of knockout populations for biochemical and functional studies.
EGFR is a transmembrane receptor tyrosine kinase activated by EGF-family ligands such as EGF, TGF-??, and HB-EGF, leading to receptor dimerization and autophosphorylation. Key adaptor proteins including GRB2, SHC, and GAB1 dock to phosphorylated tyrosines, initiating major cascades: RAS-MAPK (RAS-RAF-MEK-ERK), PI3K-AKT (PI3K-AKT-mTOR), and STAT3/STAT5 signaling. Additional pathways involve PLC??1-PKC and CBL-mediated receptor downregulation. EGFR can heterodimerize with ERBB2, ERBB3, and ERBB4. Knockout of EGFR eliminates these signaling events, abrogating ligand-driven proliferation, survival, and migration.
In the HEK293T background, endogenous EGFR expression is relatively weak, yet it is sufficient to mediate low-level signaling that can complicate interpretation of overexpressed or ligand-stimulated scenarios. The polyclonal knockout population eliminates basal receptor activity, establishing a clean genetic background for reconstitution experiments with wild-type or mutant EGFR, dissection of heterodimerization-dependent signaling, and assessment of off-target effects of EGFR-targeted therapies. Moreover, because HEK293T cells are easily transfectable and support high-level recombinant protein production, the knockout model is well-suited for biochemical reconstitution and interaction studies requiring complete absence of endogenous receptor.
This polyclonal knockout cell product supports a broad range of studies. In signal transduction research, it enables mapping of ligand-specific pathways using phospho-ERK/AKT western blotting and reconstitution with mutant EGFR. For drug development, it provides an isogenic background for testing EGFR inhibitors like erlotinib and gefitinib and identifying resistance mechanisms. Applications extend to receptor trafficking assays, co-immunoprecipitation of EGFR with GRB2/SHC, and RNA-seq profiling of EGFR-dependent transcriptomes. Cancer biology researchers can investigate EMT, proliferation, and migration in an EGFR-null context. For technical details, contact Ascent Research.
