EPCR CRISPR Activation Plasmid (h): sc-402158-ACT

PROCR encodes endothelial protein C receptor (EPCR), a surface receptor enriched on endothelial cells that binds protein C and activated protein C to regulate coagulation and vascular homeostasis. EPCR participates in anticoagulant and cytoprotective signaling through the protein C pathway, interfacing with PAR-dependent responses that influence endothelial barrier integrity and inflammatory tone. Altered EPCR expression or shedding has been associated with thromboinflammatory processes, endothelial dysfunction, and vascular complications relevant to sepsis biology and cardiovascular disease mechanisms. As a marker and regulator of endothelial state, EPCR is frequently examined in models of hemostasis, leukocyte–endothelial interactions, and microvascular injury.

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

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

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