Cyp3a11 CRISPR Activation Plasmid (m): sc-419922-ACT

Mouse Cyp3a11 encodes a cytochrome P450 monooxygenase that catalyzes oxidative metabolism of xenobiotics and endogenous substrates, contributing to hepatic clearance and chemical homeostasis. Cyp3a11 activity participates in phase I drug metabolism and interfaces with nuclear receptor signaling pathways, including PXR and CAR, which coordinate transcriptional responses to exposure and metabolic state. By shaping the biotransformation of compounds and steroids, Cyp3a11 influences pharmacokinetic variability and susceptibility to hepatotoxic injury under metabolic or inflammatory stress. Altered regulation of Cyp3a11 is therefore relevant to studies of drug–drug interactions, liver function, and gene–environment mechanisms impacting toxicology phenotypes in mouse models.

Cyp3a11 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Cyp3a11 expression without altering the underlying DNA sequence.

Cyp3a11 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Cyp3a11 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 Cyp3a11 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cyp3a11 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Cyp3a11 locus and enabling the study of Cyp3a11-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cyp3a11 pathway restoration in tumor cells with silenced or reduced Cyp3a11 expression.

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