PAI-1/SERPINE1 CRISPR Activation Plasmid (m2): sc-422287-ACT-2

Mouse Serpine1 encodes plasminogen activator inhibitor-1 (PAI-1), a secreted serpin that inhibits tissue-type and urokinase-type plasminogen activators (tPA/uPA) to restrain plasmin generation and thereby regulate fibrinolysis, pericellular proteolysis, and extracellular matrix remodeling. PAI-1 integrates cues from TGF-β, inflammatory cytokines, and oxidative stress pathways to influence cell adhesion, migration, and tissue repair, with downstream effects on wound healing and fibrotic remodeling. Dysregulated Serpine1/PAI-1 expression is linked to thrombotic and hemorrhagic phenotypes, obesity-associated metabolic inflammation, and organ fibrosis, making it a relevant target for mechanistic studies of hemostasis, vascular biology, and chronic inflammatory pathology. Gene editing or perturbation of Serpine1 in mouse models supports functional interrogation of protease–antiprotease balance, matrix turnover, and signaling networks that couple inflammation to remodeling in vivo and in cell-based assays.

PAI-1/SERPINE1 CRISPR Activation Plasmid (m2) provides a targeted, non-destructive approach to upregulating endogenous Serpine1 expression without altering the underlying DNA sequence.

PAI-1/SERPINE1 CRISPR Activation Plasmid (m2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Serpine1 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 Serpine1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PAI-1/SERPINE1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Serpine1 locus and enabling the study of PAI-1/SERPINE1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PAI-1/SERPINE1 pathway restoration in tumor cells with silenced or reduced Serpine1 expression.

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