PRX VI CRISPR Activation Plasmid (h): sc-401549-ACT

PRDX6 encodes peroxiredoxin VI (PRX VI), a bifunctional antioxidant enzyme with glutathione peroxidase activity and calcium-independent phospholipase A2 activity that supports detoxification of hydrogen peroxide and repair of oxidized membrane phospholipids. By regulating redox balance and phospholipid turnover, PRX VI influences cellular responses to oxidative stress, lipid peroxidation, and inflammatory signaling, intersecting with pathways that modulate apoptosis, proteostasis, and mitochondrial function. Altered PRDX6 expression and PRX VI activity have been associated with redox dysregulation in cancer biology, metabolic and cardiovascular disorders, and neurodegenerative processes, where oxidative injury and membrane damage are prominent features.

PRX VI CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRDX6 expression without altering the underlying DNA sequence.

PRX VI CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRDX6 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 PRDX6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PRX VI expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRDX6 locus and enabling the study of PRX VI-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PRX VI pathway restoration in tumor cells with silenced or reduced PRDX6 expression.

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