PGAM5 CRISPR Activation Plasmid (m): sc-428371-ACT

Mouse Pgam5 encodes PGAM5, a mitochondrial outer membrane Ser/Thr phosphatase that integrates metabolic status with stress signaling and mitochondrial quality control. PGAM5 participates in mitochondrial dynamics by regulating fission machinery and interfaces with mitophagy pathways, helping shape outcomes of oxidative stress and membrane depolarization. It has been linked to programmed cell death and inflammatory signaling cascades, including modulation of ASK1/JNK and related stress-activated MAPK networks. Dysregulation of PGAM5-associated pathways is relevant to models of neurodegeneration, ischemia-reperfusion injury, and immune-mediated tissue damage, making it a useful node for mechanistic studies of mitochondrial stress responses.

PGAM5 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Pgam5 expression without altering the underlying DNA sequence.

PGAM5 CRISPR Activation Plasmid (m) 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.