PTPσ CRISPR Activation Plasmid (m): sc-422531-ACT

Mouse Ptprs encodes protein tyrosine phosphatase sigma (PTPσ), a receptor-type phosphatase enriched in neural and epithelial contexts that integrates extracellular matrix cues with intracellular dephosphorylation events to shape cell adhesion, neurite outgrowth, axon guidance, and synaptic organization. PTPσ interacts with heparan sulfate and chondroitin sulfate proteoglycans, linking the pericellular matrix to cytoskeletal remodeling and signaling modules that influence growth cone behavior and circuit plasticity. Through modulation of phosphorylation-dependent pathways, Ptprs contributes to contact-dependent signaling and tissue patterning, and altered PTPσ activity has been associated in the literature with neurodevelopmental phenotypes, impaired regeneration, and dysregulated cell–cell communication. These features make Ptprs a useful target for mechanistic studies of phosphatase-controlled signaling in neuronal connectivity and microenvironmental regulation.

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

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

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