SH-PTP2 CRISPR Activation Plasmid (h): sc-400260-ACT

PTPN11 encodes SH-PTP2 (SHP2), a cytosolic protein tyrosine phosphatase containing tandem SH2 domains that couples activated receptor tyrosine kinases and cytokine receptors to downstream signaling. SHP2 is a key positive regulator of RAS–MAPK/ERK and contributes to PI3K–AKT pathway modulation, shaping cell proliferation, survival, migration, and differentiation programs. Through dephosphorylation of docking proteins and feedback nodes, SHP2 influences signal amplitude and duration across growth factor and immune signaling networks. Dysregulated PTPN11 activity and expression are associated with developmental syndromes and multiple cancer contexts, making it a common target for mechanistic studies of oncogenic signaling and resistance circuitry.

SH-PTP2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PTPN11 expression without altering the underlying DNA sequence.

SH-PTP2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PTPN11 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 PTPN11 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SH-PTP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PTPN11 locus and enabling the study of SH-PTP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SH-PTP2 pathway restoration in tumor cells with silenced or reduced PTPN11 expression.

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