PGAM5 CRISPR Activation Plasmid (h): sc-401300-ACT

PGAM5 (phosphoglycerate mutase family member 5) is an atypical mitochondrial serine/threonine phosphatase anchored to the outer mitochondrial membrane that integrates mitochondrial quality control with stress signaling. It modulates mitochondrial dynamics and mitophagy by regulating phosphorylation-dependent events on substrates involved in fission–fusion balance and can influence PINK1/Parkin-linked mitochondrial turnover. PGAM5 also interfaces with redox and cell-death signaling, including necroptosis-associated pathways, shaping cellular responses to oxidative stress and metabolic perturbation. Dysregulated PGAM5 activity or expression has been associated with mitochondrial dysfunction phenotypes implicated in neurodegeneration, cardiometabolic stress responses, and cancer biology through altered bioenergetics and stress adaptation programs.

PGAM5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PGAM5 expression without altering the underlying DNA sequence.

PGAM5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PGAM5 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 PGAM5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PGAM5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PGAM5 locus and enabling the study of PGAM5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PGAM5 pathway restoration in tumor cells with silenced or reduced PGAM5 expression.

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