Cbp CRISPR Activation Plasmid (h): sc-401415-ACT

PAG1 encodes Csk-binding protein (Cbp/PAG1), a transmembrane adaptor concentrated in lipid rafts that scaffolds signaling complexes to restrain Src family kinase activity through recruitment of Csk. By organizing phosphorylated docking sites, Cbp modulates proximal T cell receptor and B cell receptor signaling, influences downstream MAPK and PI3K pathway flux, and shapes receptor-driven activation thresholds. Altered PAG1 expression or phosphorylation has been linked to dysregulated immune signaling, aberrant proliferation, and changes in cell adhesion and migration, making it relevant to studies of inflammatory signaling and oncogenic kinase networks. In human cells, Cbp serves as a key node connecting membrane microdomain organization to checkpoint-like control of tyrosine kinase signaling.

Cbp CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PAG1 expression without altering the underlying DNA sequence.

Cbp CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PAG1 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 PAG1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cbp expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PAG1 locus and enabling the study of Cbp-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cbp pathway restoration in tumor cells with silenced or reduced PAG1 expression.

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