CYP2J2 CRISPR Activation Plasmid (h): sc-402955-ACT

CYP2J2 encodes a human cytochrome P450 epoxygenase that metabolizes arachidonic acid into epoxyeicosatrienoic acids (EETs), bioactive lipid mediators that influence vascular tone, inflammation, oxidative stress responses, and cellular survival signaling. Through its role in lipid signaling and xenobiotic metabolism, CYP2J2 contributes to endothelial and cardiac physiology and can modulate pathways linked to nitric oxide signaling, inflammatory cytokine networks, and redox homeostasis. Altered CYP2J2 expression or activity has been studied in the context of cardiovascular and metabolic phenotypes, as well as tumor biology where EET-mediated signaling can affect proliferation and migration. These features make CYP2J2 a useful target for mechanistic studies of lipid mediator biology and P450-dependent regulation of cell-state transitions.

CYP2J2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CYP2J2 expression without altering the underlying DNA sequence.

CYP2J2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CYP2J2 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 CYP2J2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CYP2J2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CYP2J2 locus and enabling the study of CYP2J2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CYP2J2 pathway restoration in tumor cells with silenced or reduced CYP2J2 expression.

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