PP1α CRISPR Activation Plasmid (h): sc-400318-ACT

PPP1CA encodes the catalytic subunit of protein phosphatase 1 alpha (PP1α), a major serine/threonine phosphatase that counterbalances kinase signaling to control protein phosphorylation homeostasis. PP1α participates in cell cycle progression, chromosome segregation, glycogen metabolism, and cytoskeletal remodeling through holoenzyme complexes formed with diverse regulatory subunits that dictate substrate specificity and subcellular localization. By modulating phosphorylation states of key signaling and structural proteins, PPP1CA influences pathways governing proliferation, DNA damage responses, and cellular stress adaptation. Dysregulated PP1α activity or targeting has been linked to aberrant phospho-signaling states observed in multiple disease contexts, supporting its use as a mechanistic node in pathway and systems-level studies.

PP1α CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PPP1CA expression without altering the underlying DNA sequence.

PP1α CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PPP1CA 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 PPP1CA transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PP1α expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PPP1CA locus and enabling the study of PP1α-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PP1α pathway restoration in tumor cells with silenced or reduced PPP1CA expression.

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