SCCA1 CRISPR Activation Plasmid (h): sc-403601-ACT

SERPINB3 encodes squamous cell carcinoma antigen 1 (SCCA1), an intracellular serine protease inhibitor of the serpin family that modulates protease-driven proteostasis and epithelial stress responses. SCCA1 can restrain cathepsin- and chymase-like protease activity, influencing lysosomal integrity, antigen processing, and apoptotic or inflammatory signaling programs in differentiated epithelia. Dysregulated SERPINB3 expression has been associated with epithelial remodeling and immune-related phenotypes observed in squamous malignancy contexts, making it a useful marker and functional node for studying tumor-associated inflammation and cell survival pathways. In cell models, SERPINB3 perturbation is commonly evaluated alongside pathways regulating oxidative stress, cytokine signaling, and epithelial differentiation.

SCCA1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SERPINB3 expression without altering the underlying DNA sequence.

SCCA1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SERPINB3 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 SERPINB3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SCCA1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SERPINB3 locus and enabling the study of SCCA1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SCCA1 pathway restoration in tumor cells with silenced or reduced SERPINB3 expression.

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