PP2A-Aβ CRISPR Activation Plasmid (h): sc-403081-ACT

PPP2R1B encodes the PP2A-Aβ scaffold subunit, a core component of protein phosphatase 2A complexes that organize catalytic and regulatory subunits to coordinate serine/threonine dephosphorylation. By shaping holoenzyme assembly, PP2A-Aβ influences signal transduction networks controlling cell-cycle progression, DNA damage responses, and cytoskeletal dynamics, with downstream effects on MAPK, AKT, and Wnt/β-catenin-associated phosphorylation states. Altered PP2A scaffold function can perturb phosphatase signaling balance and has been linked to dysregulated proliferation and genomic instability in cancer-relevant contexts. PPP2R1B is therefore frequently studied for its role in maintaining phosphoproteome homeostasis and pathway cross-talk in human cells.

PP2A-Aβ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PPP2R1B expression without altering the underlying DNA sequence.

PP2A-Aβ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PPP2R1B 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 PPP2R1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PP2A-Aβ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PPP2R1B locus and enabling the study of PP2A-Aβ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PP2A-Aβ pathway restoration in tumor cells with silenced or reduced PPP2R1B expression.

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