Endothelin-1/EDN1/ET-1 CRISPR Activation Plasmid (h): sc-400564-ACT

EDN1 encodes endothelin-1 (ET-1), a potent vasoactive peptide primarily produced by endothelial cells and processed from preproendothelin to mature ET-1. ET-1 signals through endothelin receptors (EDNRA/EDNRB) to activate G protein–coupled pathways that regulate calcium flux, MAPK signaling, cytoskeletal remodeling, and transcriptional programs controlling vasoconstriction, vascular smooth muscle proliferation, and extracellular matrix turnover. In addition to cardiovascular homeostasis, EDN1 contributes to inflammatory signaling, oxidative stress responses, and endothelial dysfunction in tissue microenvironments. Dysregulated EDN1 expression is frequently studied in pulmonary and systemic hypertension, atherosclerosis, renal disease, fibrosis, tumor angiogenesis, and other disorders where vascular remodeling and hypoxia-associated signaling are central.

Endothelin-1/EDN1/ET-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EDN1 expression without altering the underlying DNA sequence.

Endothelin-1/EDN1/ET-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EDN1 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 EDN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Endothelin-1/EDN1/ET-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EDN1 locus and enabling the study of Endothelin-1/EDN1/ET-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Endothelin-1/EDN1/ET-1 pathway restoration in tumor cells with silenced or reduced EDN1 expression.

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