The EGFR Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of HeLa cells with disrupted EGFR gene function. This product serves as a loss-of-function model to interrogate EGFR-dependent signaling in a cervical cancer context. The mixed knockout population provides a robust tool for population-based assays without clonal bias.
HeLa cells are an immortalized cervical adenocarcinoma epithelial line. They harbor HPV-18 sequences that inactivate p53 and Rb, resulting in high proliferation and genomic instability. These cells endogenously express EGFR and respond to mitogenic ligands, making them an established model for dissecting growth factor signaling in cervical and other cancers.
EGFR is a receptor tyrosine kinase that, upon binding ligands such as EGF, TGF-??, and amphiregulin, dimerizes and recruits adaptor proteins Grb2 and Shc. This activates the RAS?CRAF?CMEK?CERK cascade leading to ERK1/2 phosphorylation, and the PI3K?CAKT?CmTOR pathway, with AKT activation promoting survival. Additional signaling modules include JAK/STAT, where STAT3 phosphorylation induces transcription of targets like Cyclin D1, and PLC??/PKC, which mobilizes calcium and diacylglycerol. EGFR interacts with ErbB2/HER2 and ErbB3, and the E3 ubiquitin ligase Cbl attenuates signaling by targeting the receptor for degradation.
EGFR disruption in HeLa cells abolishes EGF-induced activation of MAPK/ERK and PI3K/AKT, reducing proliferation, survival, and migration. Given the p53/Rb deficiency, this model permits dissection of receptor?Ctumor suppressor interplay and identification of compensatory pathways. It provides a clean background for assessing EGFR-targeted agents and signaling rewiring mechanisms.
Applications include western blotting and phospho-signaling analysis using antibodies against p-ERK1/2 or p-AKT to validate pathway inactivation, RT-qPCR for transcript quantification, proliferation assays (e.g., MTT or BrdU), and migration/invasion assays in Boyden chambers to assess metastatic potential. Drug sensitivity screening against EGFR inhibitors or chemotherapeutics reveals therapeutic vulnerabilities. This polyclonal knockout model is also suitable for CRISPR-based functional genomics and synthetic lethality screens. For further details or custom inquiries, please contact Ascent Research.
