Endomucin CRISPR Activation Plasmid (h): sc-403680-ACT

EMCN encodes endomucin, a sialomucin-type glycoprotein enriched on vascular endothelial cells where it contributes to luminal surface properties, modulation of cell–cell and cell–matrix interactions, and regulation of leukocyte adhesion dynamics. Endomucin interfaces with endothelial activation programs and vascular barrier processes that coordinate inflammation and tissue perfusion. Altered EMCN expression has been associated with endothelial dysfunction signatures observed across vascular remodeling contexts, including tumor angiogenesis and inflammatory vascular disease phenotypes. As an endothelial marker and functional regulator, EMCN is frequently investigated in studies of angiogenic signaling, immune cell trafficking, and vascular niche biology.

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

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

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