ETAR CRISPR Activation Plasmid (h): sc-401000-ACT

EDNRA encodes endothelin receptor type A (ETAR), a G protein–coupled receptor that preferentially binds endothelin-1 to regulate vasoconstriction and vascular smooth muscle tone. ETAR signaling primarily engages Gq/11 to activate phospholipase C, elevate intracellular Ca2+, and stimulate PKC-dependent pathways, with downstream effects on MAPK/ERK and Rho/ROCK that influence contraction, proliferation, and extracellular matrix remodeling. In endothelial and stromal contexts, EDNRA activity intersects with hypoxia and inflammatory cues that shape angiogenic and fibrotic responses. Dysregulated EDNRA expression or signaling has been associated with cardiovascular and pulmonary vascular remodeling, systemic hypertension, and tumor-associated stromal biology, supporting its utility as a mechanistic target in pathway-focused research.

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

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

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