CYP4X1 CRISPR Activation Plasmid (m): sc-429886-ACT

Mouse Cyp4x1 encodes CYP4X1, a cytochrome P450 monooxygenase implicated in oxidative metabolism of fatty acids and other lipid-derived substrates. As part of the CYP4 family, CYP4X1 is linked to pathways that regulate lipid mediator homeostasis, redox balance, and membrane lipid remodeling, processes that can influence inflammatory signaling and vascular or neural physiology. Altered cytochrome P450 activity is frequently associated with shifts in oxidative stress responses and bioactive lipid production, making Cyp4x1 a useful target for mechanistic studies of metabolic regulation. In mouse models, modulation of CYP4X1 function can support research into how lipid oxidation pathways shape tissue-specific physiology and disease-relevant phenotypes without implying clinical outcomes.

CYP4X1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Cyp4x1 expression without altering the underlying DNA sequence.

CYP4X1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Cyp4x1 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 Cyp4x1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CYP4X1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Cyp4x1 locus and enabling the study of CYP4X1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CYP4X1 pathway restoration in tumor cells with silenced or reduced Cyp4x1 expression.

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