Periostin CRISPR Activation Plasmid (m): sc-424463-ACT

Mouse Postn encodes periostin, a secreted extracellular matrix matricellular protein that binds integrins and modulates cell–matrix adhesion, migration, and mechanotransduction. Periostin contributes to extracellular matrix remodeling, collagen fibrillogenesis, and activation of signaling pathways including FAK/PI3K/AKT, ERK/MAPK, and TGF-β-dependent programs. It is prominently expressed in connective tissues and is induced during wound repair and tissue remodeling, linking fibroblast activation to changes in stromal architecture. Dysregulated periostin expression is frequently used as a molecular indicator of fibrosis, inflammatory remodeling, and tumor-associated stroma in mouse model systems.

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

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

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