PP2Cα CRISPR Activation Plasmid (h): sc-403960-ACT

PPM1A encodes the serine/threonine phosphatase PP2Cα, a Mg²⁺/Mn²⁺-dependent PP2C family member that counterbalances stress-activated kinase signaling by dephosphorylating key pathway components. PP2Cα modulates cellular responses to genotoxic and inflammatory stimuli, with reported roles in regulating MAPK cascades, NF-κB–linked signaling outputs, and cell-cycle checkpoint control. By tuning phosphorylation-dependent signaling thresholds, PPM1A influences apoptosis, differentiation, and innate immune programs. Dysregulated PP2Cα activity or expression has been associated with altered stress adaptation and signaling phenotypes observed across multiple disease-relevant contexts, including cancer biology and immune-related pathologies.

PP2Cα CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PPM1A expression without altering the underlying DNA sequence.

PP2Cα CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PPM1A 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 PPM1A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PP2Cα expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PPM1A locus and enabling the study of PP2Cα-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PP2Cα pathway restoration in tumor cells with silenced or reduced PPM1A expression.

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