Pez CRISPR Activation Plasmid (h): sc-402896-ACT

PTPN14 encodes the protein tyrosine phosphatase non-receptor type 14 (Pez), a cytoplasmic phosphatase that integrates signals controlling cell adhesion, polarity, and contact-dependent growth. Pez participates in pathways that converge on Hippo–YAP/TAZ transcriptional regulation and coordinates actin cytoskeleton remodeling through interactions at cell–cell junctions and focal adhesion complexes. By modulating phosphorylation-dependent signaling networks, PTPN14 influences epithelial organization, migration, and mechanotransduction. Altered PTPN14 activity or expression has been linked to dysregulated proliferation and invasive phenotypes in multiple cancer-related contexts, making it relevant for studies of tumor suppressor signaling and junctional homeostasis.

Pez CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PTPN14 expression without altering the underlying DNA sequence.

Pez CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PTPN14 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 PTPN14 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Pez expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PTPN14 locus and enabling the study of Pez-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Pez pathway restoration in tumor cells with silenced or reduced PTPN14 expression.

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