EGFR CRISPR Activation Plasmid (m): sc-420131-ACT

Mouse Egfr encodes epidermal growth factor receptor (EGFR), a receptor tyrosine kinase that coordinates cellular proliferation, survival, migration, and differentiation in response to EGF-family ligands. Ligand-induced dimerization and autophosphorylation propagates signaling through canonical MAPK/ERK, PI3K–AKT–mTOR, JAK/STAT, and PLCγ/PKC pathways, integrating cues that shape developmental programs and tissue homeostasis. EGFR activity is tightly regulated by endocytosis, ubiquitin-mediated turnover, and feedback phosphorylation, and altered EGFR signaling is commonly used as a mechanistic model for aberrant growth control. In mice, Egfr is widely studied in epithelial biology, wound repair, and immune–stromal crosstalk, with pathway dysregulation informing research on oncogenic signaling, inflammation-associated remodeling, and resistance-adaptive signaling networks.

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

EGFR CRISPR Activation Plasmid (m) 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.