CYP4X1 CRISPR Activation Plasmid (h2): sc-415272-ACT-2

Human CYP4X1 encodes a cytochrome P450 monooxygenase of the CYP4 family that catalyzes oxidative metabolism of endogenous lipids, contributing to fatty acid hydroxylation and related eicosanoid-derived signaling processes. CYP4X1 activity is linked to membrane lipid homeostasis and redox-dependent regulation of cellular responses, positioning it within pathways that coordinate lipid mediator turnover and tissue-specific metabolic specialization. Dysregulated expression or altered CYP4X1-associated lipid oxidation has been investigated in the context of neurobiology and oncology, where shifts in arachidonic acid–related metabolites can influence inflammation-associated signaling and cellular proliferation programs. Gene editing of CYP4X1 supports mechanistic studies of P450-driven lipid metabolism, substrate mapping and metabolomics, and validation of pathway effects in engineered cell models relevant to human disease biology.

CYP4X1 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous CYP4X1 expression without altering the underlying DNA sequence.

CYP4X1 CRISPR Activation Plasmid (h2) 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.