Xanthine Oxidase CRISPR Activation Plasmid (h): sc-401185-ACT

XDH encodes xanthine oxidoreductase, which can function as xanthine oxidase to catalyze the terminal steps of purine catabolism, converting hypoxanthine to xanthine and xanthine to uric acid. This enzyme is a prominent source of reactive oxygen species during electron transfer reactions, linking purine metabolism to cellular redox homeostasis and oxidative stress signaling. XDH activity intersects with inflammatory and hypoxia-responsive pathways and contributes to endothelial and tissue microenvironment changes under metabolic stress. Dysregulated xanthine oxidase activity and altered urate/ROS balance are frequently investigated in contexts such as hyperuricemia-associated biology, cardiometabolic dysfunction, and ischemia-reperfusion models.

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

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

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