The EGFR Knockout HEK293 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from HEK293 human embryonic kidney cells. This model provides targeted disruption of the EGFR gene, eliminating epidermal growth factor receptor expression and creating a loss-of-function system to study EGFR-dependent signaling. The polyclonal format maintains population heterogeneity, suitable for experiments not requiring clonal isolation.
HEK293 cells are an adherent epithelial-like line transformed with adenovirus 5 DNA, widely utilized for recombinant protein expression and signal transduction studies due to their high transfection efficiency and well-characterized kinase networks. Parental HEK293 cells express functional EGFR and downstream effectors, establishing a clean background for evaluating the impact of EGFR ablation.
EGFR is a receptor tyrosine kinase that binds ligands such as EGF and TGF-alpha, triggering dimerization, autophosphorylation, and recruitment of adaptor proteins GRB2, SHC1, and GAB1. This activates the SOS1?CHRAS?CRAF1?CMAP2K1?CMAPK1/3 (ERK2/1) cascade and the PI3K?CAKT1 pathway, along with STAT3 and PLCG1 signaling. Downstream, FOS, JUN, MYC, and CCND1 are transcriptionally induced. In knockout cells, the absence of EGFR prevents ligand-induced phosphorylation of MAPK1/3 and AKT1, and disrupts formation of complexes with GRB2 and CBL, abrogating multiple branches of EGF-dependent signal transduction.
Endogenous EGFR in HEK293 contributes to basal and ligand-stimulated MAPK/ERK and PI3K/AKT activation. The knockout polyclonal cells enable clear dissection of EGFR-specific functions versus contributions from other ErbB receptors or parallel pathways. This system is ideal for studying EGFR crosstalk, receptor trafficking, and feedback loops, and for distinguishing direct from indirect effects in EGF-dependent processes.
This product supports diverse applications, including EGFR signaling mechanism studies, inhibitor validation (e.g., erlotinib), and receptor trafficking analysis. Typical assays include Western blotting for phospho-MAPK1/3 and AKT1 after EGF stimulation, RT-qPCR for EGR1 and FOS, co-immunoprecipitation of GRB2, flow cytometry for surface EGFR, and proliferation assays. The polyclonal population is suited for drug sensitivity screens and compensatory signaling exploration. For more information, contact Ascent Research.
