PRX V CRISPR Activation Plasmid (h): sc-402681-ACT

Peroxiredoxin 5 (PRDX5; PRX V) is a thioredoxin-dependent peroxidase that detoxifies hydrogen peroxide and organic hydroperoxides, helping maintain cellular redox homeostasis across mitochondria, peroxisomes, and the cytosol. By limiting oxidative damage to lipids, proteins, and DNA, PRX V influences mitochondrial function, stress-adaptive signaling, and redox-sensitive pathways that shape inflammation and cell survival. Altered PRDX5 expression or activity has been linked to oxidative stress phenotypes observed in neurodegeneration, metabolic dysfunction, cardiovascular injury models, and tumor biology, where reactive oxygen species modulate proliferation and apoptosis. As a redox regulator, PRDX5 is frequently studied for its role in controlling reactive oxygen species thresholds that impact innate immune responses and mitochondrial integrity.

PRX V CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRDX5 expression without altering the underlying DNA sequence.

PRX V CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRDX5 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 PRDX5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PRX V expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRDX5 locus and enabling the study of PRX V-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PRX V pathway restoration in tumor cells with silenced or reduced PRDX5 expression.

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