The EGFR Knockout 143B Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human osteosarcoma cell line 143B, enabling study of EGFR loss-of-function without single-clone selection bias. The polyclonal format preserves genetic diversity, making it suitable for functional genomics, pathway analysis, and anti-cancer drug screening.
The 143B cell line is a widely used human osteosarcoma model with epithelial morphology, employed to investigate progression, metastasis, and tumorigenesis. These cells harbor oncogenic properties and provide a robust platform for studying signaling networks in aggressive bone cancer. CRISPR-mediated EGFR disruption in this background offers a physiologically relevant system to dissect receptor tyrosine kinase signaling in a cancer type where EGFR contributes to malignancy.
EGFR is a receptor tyrosine kinase that, upon binding ligands EGF or TGF-??, activates multiple cascades controlling proliferation, survival, and migration. Activated EGFR recruits GRB2 and SHC, engaging SOS to stimulate RAS, which triggers RAF/MEK/ERK signaling. Concurrent GAB1-mediated PI3K/AKT and PLC??1/PKC pathways converge on effectors ERK1/2, AKT, and STAT3, regulating transcription factors c-Fos and c-Jun to drive expression of cyclin D1, MYC, and MMPs. The network is modulated by CBL-mediated receptor internalization.
In osteosarcoma, aberrant EGFR signaling promotes proliferation, apoptotic resistance, and invasiveness. EGFR disruption in 143B cells eliminates EGF-induced MAPK/ERK and PI3K/AKT activation, attenuating proliferative and survival signals. This knockout enables precise interrogation of EGFR dependency in transcriptional reprogramming and cell cycle progression. Loss of EGFR-driven MMP expression provides a model for studying metastasis suppression. Given the rarity of EGFR mutations in osteosarcoma, this knockout serves as a clean system for evaluating EGFR-targeted therapies and resistance.
The polyclonal pool supports diverse applications: Western blotting for EGFR and phospho-ERK/AKT, MTT/BrdU proliferation assays, wound healing and transwell invasion tests, colony formation, and RT-qPCR of downstream targets. Immunofluorescence confirms EGFR loss at single-cell resolution, and xenograft studies assess tumor growth impairment. For inquiries or to discuss integrating these cells into osteosarcoma and EGFR research, contact Ascent Research.
