Epiregulin CRISPR Activation Plasmid (h): sc-404273-ACT

EREG encodes epiregulin, a secreted EGFR/ERBB family ligand that modulates epithelial and stromal signaling in a context-dependent manner. By engaging EGFR and ERBB4, epiregulin activates downstream MAPK/ERK, PI3K–AKT, and JAK/STAT signaling cascades that regulate proliferation, differentiation, migration, and inflammatory crosstalk. EREG expression is frequently inducible by cytokines and growth factors and contributes to wound repair programs, tissue remodeling, and paracrine communication within the tumor microenvironment. Dysregulated EREG has been associated with oncogenic signaling, invasion phenotypes, and altered immune–epithelial interactions, supporting its use as a mechanistic node in cancer biology and inflammation research.

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

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

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