PTPκ CRISPR Activation Plasmid (h): sc-402287-ACT

PTPRK encodes protein tyrosine phosphatase receptor type K (PTPκ), a receptor-like phosphatase that modulates phosphorylation-dependent signaling at the plasma membrane and cell–cell junctions. By dephosphorylating substrates involved in adhesion and growth factor signaling, PTPκ contributes to regulation of epithelial integrity, contact inhibition, and cytoskeletal organization. PTPRK activity intersects with pathways linked to adherens junction dynamics and receptor tyrosine kinase outputs, influencing proliferation and migration programs. Altered PTPRK expression or function has been associated with dysregulated signaling networks observed in several disease contexts, supporting its utility as a mechanistic target in cell signaling and junction biology studies.

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

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

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