CYPOR CRISPR Activation Plasmid (m): sc-422345-ACT

Mouse Por encodes cytochrome P450 oxidoreductase (CYPOR), an ER membrane flavoprotein that transfers electrons from NADPH to microsomal cytochrome P450 enzymes and other redox partners. By enabling P450 catalytic cycles, CYPOR supports oxidative metabolism of endogenous lipids and steroids as well as xenobiotic biotransformation, linking Por activity to hepatic detoxification, redox homeostasis, and ER-associated metabolic networks. Altered CYPOR function perturbs P450-dependent pathways and can shift reactive oxygen species balance, making Por a key variable in models of metabolic stress and pharmacology-related phenotypes. In biomedical research, Por is commonly studied to disentangle P450 electron transfer requirements from substrate-specific effects and to map genotype-to-metabolite relationships across tissues.

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

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

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