PCYOX1 CRISPR Activation Plasmid (h): sc-407075-ACT

PCYOX1 (prenylcysteine oxidase 1) encodes a flavin-dependent oxidase that catalyzes the oxidative degradation of prenylcysteines generated during turnover of prenylated proteins, contributing to clearance of lipid-modified cysteine metabolites. Through this catabolic step, PCYOX1 links protein prenylation dynamics to cellular redox balance and metabolic homeostasis, with downstream effects on oxidative stress signaling and inflammatory responses. Altered PCYOX1 expression and activity have been associated with pathways relevant to cardiometabolic traits, vascular biology, and lipid handling, supporting its study in models of atherosclerosis and related inflammatory states. In human cells, PCYOX1 perturbation can be used to probe how prenylation byproducts shape mitochondrial function, reactive oxygen species production, and stress-adaptive transcriptional programs.

PCYOX1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PCYOX1 expression without altering the underlying DNA sequence.

PCYOX1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PCYOX1 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 PCYOX1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PCYOX1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PCYOX1 locus and enabling the study of PCYOX1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PCYOX1 pathway restoration in tumor cells with silenced or reduced PCYOX1 expression.

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