CYP2E1 CRISPR Activation Plasmid (m): sc-419917-ACT

Mouse Cyp2e1 encodes the cytochrome P450 enzyme CYP2E1, a microsomal monooxygenase that oxidizes small organic substrates including ethanol, acetone, and other low–molecular weight xenobiotics. CYP2E1 activity contributes to phase I metabolism and can elevate reactive oxygen species production, linking it to oxidative stress, lipid peroxidation, and endoplasmic reticulum stress responses in hepatocytes. In the liver, CYP2E1 intersects with pathways governing xenobiotic clearance, redox homeostasis, and inflammatory signaling. Altered Cyp2e1 expression or activity is frequently studied in models of toxicant-induced liver injury, metabolic dysfunction, and alcohol-associated liver pathology to understand how metabolic activation and ROS drive tissue damage.

CYP2E1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Cyp2e1 expression without altering the underlying DNA sequence.

CYP2E1 CRISPR Activation Plasmid (m) 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.