plasma kallikrein CRISPR Activation Plasmid (h): sc-402237-ACT

KLKB1 encodes plasma kallikrein, a serine protease that functions in the contact activation system to regulate intrinsic coagulation, fibrinolysis, and inflammatory signaling. Upon activation, plasma kallikrein cleaves high-molecular-weight kininogen to release bradykinin, linking KLKB1 activity to vascular permeability, edema biology, and cytokine-modulating pathways. It also participates in reciprocal activation with coagulation factor XII and can influence protease-activated receptor signaling and complement-related cascades through proteolytic cross-talk. Dysregulated kallikrein–kinin system activity is implicated in disorders of bradykinin-mediated inflammation and angioedema-like phenotypes, making KLKB1 a useful target for mechanistic studies of hemostasis–inflammation coupling.

plasma kallikrein CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KLKB1 expression without altering the underlying DNA sequence.

plasma kallikrein CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KLKB1 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 KLKB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous plasma kallikrein expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KLKB1 locus and enabling the study of plasma kallikrein-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of plasma kallikrein pathway restoration in tumor cells with silenced or reduced KLKB1 expression.

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