PRX II CRISPR Activation Plasmid (h): sc-401330-ACT

PRDX2 encodes human peroxiredoxin-2 (PRX II), a cytosolic thiol-dependent peroxidase that reduces hydrogen peroxide and organic hydroperoxides to help maintain cellular redox homeostasis. By controlling peroxide tone and thiol signaling, PRX II influences ROS-sensitive pathways that regulate proliferation, apoptosis, and inflammatory responses, including redox modulation of MAPK, NF-κB, and Nrf2-associated programs. PRX II also contributes to antioxidant defense in erythrocytes and other high-oxidative-burden contexts, linking its activity to oxidative stress phenotypes. Altered PRDX2 expression or PRX II function has been associated with redox imbalance observed across cancer biology, neurodegeneration, and cardiovascular and inflammatory disease models, supporting its use in mechanistic studies of oxidative signaling.

PRX II CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRDX2 expression without altering the underlying DNA sequence.

PRX II CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRDX2 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 PRDX2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PRX II expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRDX2 locus and enabling the study of PRX II-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PRX II pathway restoration in tumor cells with silenced or reduced PRDX2 expression.

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