PRX V CRISPR Activation Plasmid (m): sc-424925-ACT

Mouse Prdx5 encodes peroxiredoxin V (PRX V), an atypical 2-Cys peroxiredoxin that reduces hydrogen peroxide and organic hydroperoxides to help maintain cellular redox homeostasis. PRX V supports antioxidant defense across multiple subcellular compartments and modulates peroxide-dependent signaling that influences mitochondrial function, innate immune responses, and apoptosis. By shaping reactive oxygen species flux, PRX V intersects with redox-regulated pathways controlling metabolism, stress adaptation, and inflammatory signaling. Altered PRDX5/PRX V activity has been associated with oxidative-stress phenotypes relevant to neurodegeneration, metabolic dysfunction, and inflammatory injury models, making it useful for mechanistic studies of redox biology.

PRX V CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Prdx5 expression without altering the underlying DNA sequence.

PRX V CRISPR Activation Plasmid (m) 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.