PP2A-B56-β CRISPR Activation Plasmid (h): sc-403233-ACT

PPP2R5B encodes the PP2A regulatory subunit B56β, a member of the B56 family that directs protein phosphatase 2A holoenzyme assembly and substrate specificity. By targeting PP2A to defined complexes, PP2A-B56β contributes to dynamic control of phosphorylation-dependent signaling, including cell-cycle progression, DNA damage responses, and mitotic checkpoint regulation. PP2A-B56β–guided dephosphorylation also intersects with PI3K/AKT and MAPK pathway nodes through regulation of kinase and scaffold substrates, shaping proliferation and stress-adaptation programs. Dysregulated PP2A-B56β expression or function has been associated with aberrant signaling and genomic instability phenotypes that are frequently studied in cancer biology and other proliferative disorders.

PP2A-B56-β CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PPP2R5B expression without altering the underlying DNA sequence.

PP2A-B56-β CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PPP2R5B 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 PPP2R5B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PP2A-B56-β expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PPP2R5B locus and enabling the study of PP2A-B56-β-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PP2A-B56-β pathway restoration in tumor cells with silenced or reduced PPP2R5B expression.

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