GPVI CRISPR Activation Plasmid (h): sc-404055-ACT

GP6 encodes glycoprotein VI (GPVI), a platelet-specific immunoglobulin superfamily receptor that mediates adhesion and activation in response to collagen and related ligands exposed at sites of vascular injury. GPVI signals through the FcRγ chain and ITAM-dependent pathways to engage Src and Syk kinases, triggering PLCγ2 activation, intracellular calcium mobilization, integrin activation, secretion, and cytoskeletal remodeling. These processes coordinate thrombus formation and platelet–vessel wall interactions and intersect with broader hemostatic and inflammatory signaling networks. Altered GPVI axis activity is studied in the context of platelet function disorders, thrombo-inflammatory mechanisms, and vascular disease biology.

GPVI CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GP6 expression without altering the underlying DNA sequence.

GPVI CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GP6 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 GP6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPVI expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GP6 locus and enabling the study of GPVI-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPVI pathway restoration in tumor cells with silenced or reduced GP6 expression.

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