PTP1B CRISPR Activation Plasmid (h): sc-400732-ACT

PTPN1 encodes protein tyrosine phosphatase 1B (PTP1B), a non-receptor phosphatase anchored to the cytosolic face of the endoplasmic reticulum that attenuates signaling by dephosphorylating activated receptor tyrosine kinases and associated substrates. PTP1B modulates insulin and leptin receptor pathways and intersects with growth factor–driven cascades including MAPK/ERK and PI3K–AKT, shaping cellular programs such as glucose homeostasis, proliferation, and survival. Through its role in tuning phosphorylation-dependent signaling networks, altered PTPN1/PTP1B activity has been linked to metabolic dysregulation and to oncogenic signaling contexts where receptor and downstream kinase activity are rewired. These properties make PTPN1 a useful target for mechanistic studies of phosphotyrosine signaling, feedback control, and pathway cross-talk in human cell models.

PTP1B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PTPN1 expression without altering the underlying DNA sequence.

PTP1B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PTPN1 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 PTPN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PTP1B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PTPN1 locus and enabling the study of PTP1B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PTP1B pathway restoration in tumor cells with silenced or reduced PTPN1 expression.

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