Endothelin-2/EDN2/ET-2 CRISPR Activation Plasmid (h): sc-403871-ACT

EDN2 encodes endothelin-2 (ET-2), a potent vasoactive peptide processed from a prepropeptide and acting primarily through endothelin receptors (EDNRA/EDNRB) to regulate smooth muscle contraction, endothelial signaling, and paracrine control of tissue perfusion. ET-2 signaling engages G protein–coupled pathways that modulate intracellular calcium, MAPK/ERK, and PLC/PKC cascades, influencing proliferation, migration, and inflammatory mediator production. Beyond vascular biology, EDN2 is implicated in reproductive and ovarian follicular dynamics, hypoxia-responsive transcriptional programs, and stromal remodeling. Dysregulated endothelin axis activity, including altered EDN2 expression, has been associated with fibrotic processes, pulmonary and cardiovascular pathophysiology, and tumor microenvironment signaling relevant to cancer biology.

Endothelin-2/EDN2/ET-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EDN2 expression without altering the underlying DNA sequence.

Endothelin-2/EDN2/ET-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EDN2 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 EDN2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Endothelin-2/EDN2/ET-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EDN2 locus and enabling the study of Endothelin-2/EDN2/ET-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Endothelin-2/EDN2/ET-2 pathway restoration in tumor cells with silenced or reduced EDN2 expression.

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