TC-PTP CRISPR Activation Plasmid (h): sc-403071-ACT

Human PTPN2 encodes T cell protein tyrosine phosphatase (TC-PTP), a non-receptor PTP that dephosphorylates signaling intermediates to constrain cytokine and growth factor responses. TC-PTP is a key negative regulator of JAK/STAT signaling, modulating phosphorylation of STAT family members and receptor-associated kinases to shape transcriptional programs controlling immune homeostasis, proliferation, and cellular stress responses. Through cross-talk with pathways downstream of receptor tyrosine kinases and inflammatory cues, PTPN2 helps calibrate thresholds for activation and feedback inhibition. Genetic and functional studies link altered PTPN2 activity or expression to dysregulated inflammatory signaling and susceptibility to immune-mediated and metabolic phenotypes, supporting its broad relevance in disease mechanism research.

TC-PTP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PTPN2 expression without altering the underlying DNA sequence.

TC-PTP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PTPN2 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 PTPN2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TC-PTP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PTPN2 locus and enabling the study of TC-PTP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TC-PTP pathway restoration in tumor cells with silenced or reduced PTPN2 expression.

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