Sulfiredoxin CRISPR Activation Plasmid (h): sc-403023-ACT

SRXN1 encodes sulfiredoxin, an ATP-dependent oxidoreductase that restores hyperoxidized peroxiredoxins (Prx-SO2H) to their active thiol form, sustaining peroxidase activity during oxidative stress. By maintaining peroxiredoxin cycling, sulfiredoxin supports redox homeostasis and influences ROS-sensitive signaling pathways, including Nrf2/ARE-driven antioxidant responses and downstream modulation of MAPK and NF-κB signaling. SRXN1 activity is linked to cellular adaptation under hypoxia, inflammation, and proteotoxic stress, and altered expression has been reported across contexts of cancer biology, neurodegeneration, and metabolic dysfunction where redox imbalance is a key feature. These properties make SRXN1 a useful node for studying thiol redox regulation, oxidative damage repair, and stress-induced transcriptional programs in human cells.

Sulfiredoxin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SRXN1 expression without altering the underlying DNA sequence.

Sulfiredoxin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SRXN1 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 SRXN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Sulfiredoxin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SRXN1 locus and enabling the study of Sulfiredoxin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Sulfiredoxin pathway restoration in tumor cells with silenced or reduced SRXN1 expression.

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