EGFR CRISPR Activation Plasmid (h): sc-400015-ACT

Epidermal growth factor receptor (EGFR) is a human receptor tyrosine kinase that initiates signaling cascades controlling cell proliferation, survival, differentiation, and migration following ligand-induced dimerization and autophosphorylation. EGFR activates core pathways including RAS–RAF–MEK–ERK, PI3K–AKT–mTOR, PLCγ–PKC, and JAK–STAT, integrating extracellular growth cues with transcriptional and metabolic programs. Dysregulated EGFR expression or signaling is frequently associated with altered epithelial homeostasis and oncogenic phenotypes, making it a central node in growth factor receptor biology. EGFR is also widely used as a model receptor for studying receptor trafficking, endocytosis, feedback regulation, and pathway cross-talk in human cell systems.

EGFR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EGFR expression without altering the underlying DNA sequence.

EGFR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EGFR locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the EGFR transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EGFR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EGFR locus and enabling the study of EGFR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EGFR pathway restoration in tumor cells with silenced or reduced EGFR expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.