PP2Cγ CRISPR Activation Plasmid (h): sc-404206-ACT

PPM1G encodes the serine/threonine protein phosphatase PP2Cγ, a PP2C family enzyme that regulates phosphorylation-dependent signaling and nuclear processes. PP2Cγ has been linked to control of gene expression programs through effects on chromatin-associated factors, RNA processing, and cell-cycle–related phosphorylation networks, helping tune stress responses and proliferation. By counterbalancing kinase activity, PPM1G contributes to maintenance of phospho-protein homeostasis that influences genome integrity and transcriptional outputs. Altered regulation of PP2Cγ-associated pathways has been observed in contexts relevant to oncogenic signaling, DNA damage responses, and aberrant cell-state transitions, making it a useful node for mechanistic studies.

PP2Cγ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PPM1G expression without altering the underlying DNA sequence.

PP2Cγ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PPM1G 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 PPM1G transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PP2Cγ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PPM1G locus and enabling the study of PP2Cγ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PP2Cγ pathway restoration in tumor cells with silenced or reduced PPM1G expression.

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