ETBR CRISPR Activation Plasmid (m): sc-420112-ACT

Ednrb encodes endothelin receptor type B (ETBR), a G protein–coupled receptor that binds endothelins to regulate vasomotor signaling, neural crest–derived cell development, and melanocyte and enteric nervous system biology. ETBR activation engages canonical GPCR pathways including Gq/PLCβ/Ca2+ mobilization and MAPK signaling, shaping cell migration, differentiation, and tissue patterning. In mouse models, Ednrb function is tightly linked to neural crest lineage specification and peripheral nervous system formation, with perturbations associated with aganglionosis and pigmentation defects. These properties make ETBR a relevant node for studying developmental signaling, cell–cell communication, and endothelin pathway crosstalk in physiological and disease-relevant contexts.

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

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

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