CYP2E1 CRISPR Activation Plasmid (h): sc-401357-ACT

Human CYP2E1 encodes a cytochrome P450 monooxygenase that catalyzes oxidation of small, low–molecular weight substrates, including ethanol, acetone, and diverse xenobiotics, shaping hepatic metabolic capacity. Through NADPH-dependent electron transfer and coupling to redox homeostasis, CYP2E1 activity can elevate reactive oxygen species and lipid peroxidation, linking it to cellular stress responses and inflammatory signaling. CYP2E1 is embedded in drug metabolism and chemical carcinogenesis pathways and is frequently studied in the context of liver injury, steatohepatitis, and toxin-mediated hepatotoxicity. Variation in CYP2E1 expression also influences inter-individual differences in xenobiotic clearance and metabolite formation relevant to pharmacology and environmental exposure biology.

CYP2E1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CYP2E1 expression without altering the underlying DNA sequence.

CYP2E1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CYP2E1 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 CYP2E1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CYP2E1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CYP2E1 locus and enabling the study of CYP2E1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CYP2E1 pathway restoration in tumor cells with silenced or reduced CYP2E1 expression.

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