Pi16 CRISPR Activation Plasmid (m): sc-428732-ACT

Mouse peptidase inhibitor 16 (Pi16) encodes a secreted serine protease inhibitor–like protein implicated in regulating extracellular proteolysis and pericellular matrix remodeling. Pi16 expression is enriched in stromal compartments and mesenchymal lineages, linking it to fibroblast biology, tissue repair programs, and modulation of cell–matrix interactions that influence migration and differentiation. Through its effects on protease activity and extracellular signaling availability, Pi16 is relevant to pathways governing inflammation-associated remodeling and fibrotic responses. Dysregulated stromal remodeling and altered protease/inhibitor balance are recurrent features of cardiovascular and pulmonary disease models, positioning Pi16 as a useful node for mechanistic studies of microenvironmental regulation.

Pi16 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Pi16 expression without altering the underlying DNA sequence.

Pi16 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Pi16 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 Pi16 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Pi16 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Pi16 locus and enabling the study of Pi16-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Pi16 pathway restoration in tumor cells with silenced or reduced Pi16 expression.

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