Maspin CRISPR Activation Plasmid (h): sc-416693-ACT

Human SERPINB5 encodes maspin, a non-inhibitory serpin family member implicated in regulating epithelial cell adhesion, motility, and extracellular matrix interactions. Maspin has been linked to modulation of plasminogen activation, cell–matrix signaling, and cytoskeletal dynamics, influencing processes such as invasion, migration, and tissue remodeling. Its expression and subcellular localization are frequently altered across tumor types, where maspin is studied as a context-dependent regulator of tumor progression and metastasis-associated phenotypes. As a result, SERPINB5 is widely used to interrogate pathways connecting epithelial differentiation, stress responses, and microenvironmental cues in cancer biology.

Maspin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SERPINB5 expression without altering the underlying DNA sequence.

Maspin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SERPINB5 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 SERPINB5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Maspin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SERPINB5 locus and enabling the study of Maspin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Maspin pathway restoration in tumor cells with silenced or reduced SERPINB5 expression.

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