PAI-2 CRISPR Activation Plasmid (h): sc-402054-ACT

SERPINB2 encodes plasminogen activator inhibitor-2 (PAI-2), a serine protease inhibitor that modulates pericellular proteolysis by inhibiting urokinase-type plasminogen activator (uPA/PLAU), thereby influencing plasmin generation and extracellular matrix remodeling. PAI-2 is induced by inflammatory cues and is frequently studied in monocytes/macrophages, epithelial cells, and tumor microenvironments where it intersects with fibrinolysis, wound response, and cytokine-driven signaling networks. Through regulation of uPA/uPAR-linked processes, SERPINB2 impacts cell migration, adhesion, and protease-dependent signaling cascades relevant to tissue remodeling and immune activation. Altered SERPINB2/PAI-2 expression has been associated with inflammatory pathologies and cancer-related phenotypes, supporting its use as a mechanistic node in studies of proteostasis and inflammation-coupled remodeling.

PAI-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SERPINB2 expression without altering the underlying DNA sequence.

PAI-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SERPINB2 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 SERPINB2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PAI-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SERPINB2 locus and enabling the study of PAI-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PAI-2 pathway restoration in tumor cells with silenced or reduced SERPINB2 expression.

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