P-Selectin/CD62P/SELP CRISPR Activation Plasmid (h): sc-400441-ACT

SELP encodes P-selectin (CD62P), an inducible adhesion receptor stored in platelet α-granules and endothelial Weibel–Palade bodies and rapidly displayed on the cell surface upon activation. Surface P-selectin binds PSGL-1 on leukocytes to initiate tethering and rolling, coordinating early leukocyte recruitment during inflammation and linking vascular injury to thromboinflammatory responses. Through regulation of platelet–leukocyte and endothelium–leukocyte interactions, SELP influences coagulation-associated signaling, cytokine-driven endothelial activation, and remodeling of the vascular microenvironment. Dysregulated P-selectin expression and activity are associated with inflammatory vascular pathology, thrombosis-related processes, and tumor-associated platelet interactions that impact metastasis biology.

P-Selectin/CD62P/SELP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SELP expression without altering the underlying DNA sequence.

P-Selectin/CD62P/SELP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SELP 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 SELP transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous P-Selectin/CD62P/SELP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SELP locus and enabling the study of P-Selectin/CD62P/SELP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of P-Selectin/CD62P/SELP pathway restoration in tumor cells with silenced or reduced SELP expression.

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