Serpinb3a CRISPR Activation Plasmid (m): sc-422808-ACT

Mouse Serpinb3a encodes a clade B serine protease inhibitor (serpin) implicated in regulating intracellular protease activity, cellular stress responses, and epithelial homeostasis. As an intracellular serpin, Serpinb3a can influence protease-dependent processes linked to inflammation, tissue remodeling, and barrier integrity, intersecting with pathways that modulate apoptosis, oxidative stress adaptation, and immune signaling. Altered expression of serpin family members has been associated with dysregulated inflammatory microenvironments and epithelial injury phenotypes, making Serpinb3a a useful molecular node for studying context-dependent stress and proteostasis programs. In mouse models, Serpinb3a expression dynamics are frequently evaluated in studies of tissue damage, fibrotic remodeling, and immune-mediated pathology where protease–antiprotease balance is a key determinant of cellular outcomes.

Serpinb3a CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Serpinb3a expression without altering the underlying DNA sequence.

Serpinb3a CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Serpinb3a 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 Serpinb3a transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Serpinb3a expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Serpinb3a locus and enabling the study of Serpinb3a-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Serpinb3a pathway restoration in tumor cells with silenced or reduced Serpinb3a expression.

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