ADAMTS-13 CRISPR Activation Plasmid (h): sc-402782-ACT

ADAMTS13 encodes ADAMTS-13, a secreted zinc-dependent metalloprotease that specifically cleaves ultra-large von Willebrand factor (VWF) multimers, thereby regulating platelet adhesion and thrombus formation under high shear stress. By controlling VWF multimer size and distribution, ADAMTS-13 modulates hemostasis-related extracellular proteolysis and endothelial-platelet interactions within the vascular microenvironment. Reduced ADAMTS-13 activity or dysregulated VWF processing is associated with microangiopathic thrombotic disorders, making ADAMTS13 a key locus for studying coagulation biology and vascular inflammation. ADAMTS-13 also provides a functional entry point for interrogating pathways linking endothelial activation, shear-dependent mechanobiology, and plasma protease networks.

ADAMTS-13 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADAMTS13 expression without altering the underlying DNA sequence.

ADAMTS-13 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADAMTS13 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 ADAMTS13 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ADAMTS-13 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADAMTS13 locus and enabling the study of ADAMTS-13-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ADAMTS-13 pathway restoration in tumor cells with silenced or reduced ADAMTS13 expression.

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