PR3 CRISPR Activation Plasmid (h): sc-402550-ACT

PRTN3 encodes proteinase 3 (PR3), a neutrophil serine protease stored in azurophilic granules and mobilized to phagosomes and the cell surface during innate immune activation. PR3 contributes to antimicrobial defense and inflammatory signaling by proteolytic processing of extracellular matrix components, cytokines, and chemokine networks, interfacing with degranulation, NET formation, and leukocyte trafficking pathways. Dysregulated PR3 activity and aberrant immune recognition are linked to inflammatory tissue injury and serve as important mechanistic readouts in studies of neutrophil-driven pathology, including autoimmune vasculitic processes. PRTN3 expression dynamics are therefore frequently interrogated in myeloid differentiation models, inflammatory signaling assays, and protease-substrate pathway mapping.

PR3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRTN3 expression without altering the underlying DNA sequence.

PR3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRTN3 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 PRTN3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PR3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRTN3 locus and enabling the study of PR3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PR3 pathway restoration in tumor cells with silenced or reduced PRTN3 expression.

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