Amphiregulin CRISPR Activation Plasmid (h): sc-400987-ACT

AREG encodes amphiregulin, a secreted ligand of the epidermal growth factor receptor (EGFR) that regulates epithelial cell proliferation, survival, and differentiation through downstream MAPK/ERK, PI3K/AKT, and JAK/STAT signaling. Amphiregulin is inducible by inflammatory cues and contributes to tissue repair, stromal–epithelial communication, and remodeling of the local microenvironment. Dysregulated AREG–EGFR signaling has been linked to altered immune responses and aberrant growth programs observed across multiple disease contexts, including cancer biology and chronic inflammatory states. As a readout of EGFR pathway activity, AREG expression is frequently used to study ligand-dependent signaling dynamics, feedback regulation, and paracrine signaling networks.

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

Amphiregulin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AREG 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 AREG transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Amphiregulin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AREG locus and enabling the study of Amphiregulin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Amphiregulin pathway restoration in tumor cells with silenced or reduced AREG expression.

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