Pepsin C CRISPR Activation Plasmid (h): sc-404137-ACT

PGC encodes pepsin C (progastricsin), an aspartic endopeptidase primarily produced by gastric chief cells and secreted as an inactive zymogen that becomes proteolytically active in acidic environments. Once activated, pepsin C contributes to luminal protein processing and supports gastric mucosal physiology through regulated secretion and protease activity within the digestive enzyme network. Aberrant PGC expression patterns are widely used as indicators of altered gastric epithelial differentiation and mucosal remodeling, linking this gene to research on gastric inflammation, metaplasia, and neoplastic transformation. As a stomach-enriched protease gene, PGC is also informative for studying epithelial lineage programs and proteostasis changes associated with gastric disease biology.

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

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

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