h/m EGFR shRNA Lentivirus

Background

EGFR encodes the epidermal growth factor receptor, a receptor tyrosine kinase that binds ligands such as EGF and TGF-α. Ligand binding triggers receptor dimerization and autophosphorylation, activating downstream RAS/MAPK, PI3K/AKT, and STAT signaling cascades that control cell proliferation, survival, migration, and differentiation. EGFR is a well-established oncogene: amplification, overexpression, and activating mutations drive many epithelial cancers, including lung, colorectal, and head and neck tumors. EGFR is one of the most prominent targets of cancer therapy, with both antibody and small-molecule inhibitors in clinical use, making it a central focus of cancer and cell biology research.

Product Description & Applications

The h/m EGFR shRNA Lentivirus delivers a validated short hairpin RNA targeting human and mouse EGFR for stable RNA interference. The shRNA is expressed from a U6 promoter in a third-generation, self-inactivating lentiviral backbone, with a co-expressed fluorescent reporter (GFP or RFP, optionally with luciferase) and optional blasticidin or puromycin selection. Particles are ultra-purified and concentrated by PEG precipitation and sucrose gradient centrifugation and efficiently transduce difficult-to-transfect cells, including primary and thawed cells. The shRNA is validated for at least 70% knockdown using a fluorescence-based assay.

A shRNA set option supplies a mix of two validated shRNAs plus a scrambled control for loss-of-function studies of EGFR signaling and tumor cell biology.

About This Product

This validated shRNA lentivirus targeting EGFR (NCBI Accession: NM_005228.5) delivers a 19–20 bp shRNA from a third-generation, self-inactivating lentiviral backbone. Expression is driven from a U6 Pol III promoter, with a constitutively expressed fluorescent reporter (GFP, GFP/Luc, RFP, RFP/Luc) and antibiotic selection marker (Blasticidin, Puromycin) co-expressed from the same vector. VSV-G pseudotyping enables broad cell tropism, including primary, suspension, and cryopreserved cell types.

Knockdown is validated using a proprietary bicistronic fluorescence assay in which the target mRNA is co-expressed fused to RFP alongside the shRNA-GFP construct. At least 70% reduction in RFP signal in GFP-positive cells confirms on-target activity — a more direct functional readout than transcript-level qPCR. Polyclonal stable lines can be generated by antibiotic selection within 10 days, preserving parental cell heterogeneity compared to single-clone CRISPR approaches.