Fap-1 CRISPR Activation Plasmid (h): sc-401758-ACT

PTPN13 encodes the large non-receptor protein tyrosine phosphatase Fap-1 (also known as PTP-BAS), a multi-PDZ domain scaffold that integrates phosphatase activity with protein–protein interactions at the plasma membrane and cytoskeleton. Fap-1 modulates signaling outputs by dephosphorylating key substrates and organizing receptor-associated complexes, influencing processes such as apoptosis regulation, cell adhesion, and migration. Through interactions with pathways linked to receptor signaling and junctional dynamics, PTPN13 helps shape cellular responses to growth and stress cues. Altered PTPN13/Fap-1 expression or signaling balance has been associated with dysregulated survival and invasive phenotypes reported in diverse disease contexts, making it a useful node for mechanistic studies of signaling control.

Fap-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PTPN13 expression without altering the underlying DNA sequence.

Fap-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PTPN13 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 PTPN13 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Fap-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PTPN13 locus and enabling the study of Fap-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Fap-1 pathway restoration in tumor cells with silenced or reduced PTPN13 expression.

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