CYP4F8 CRISPR Activation Plasmid (h): sc-411598-ACT

Human CYP4F8 encodes a cytochrome P450 monooxygenase that contributes to oxidative metabolism of fatty acid–derived mediators, supporting regulation of local lipid signaling and xenobiotic processing. As part of the broader CYP4 family, CYP4F8 is linked to ω-hydroxylation reactions that can influence eicosanoid turnover and inflammatory tone, intersecting with pathways governing epithelial homeostasis and cellular stress responses. Variation in CYP450 activity and expression is frequently investigated in the context of inter-individual differences in metabolism and susceptibility to inflammation-associated phenotypes, making CYP4F8 a useful target in mechanistic studies. In vitro models leveraging CYP4F8 modulation can help clarify how lipid mediator metabolism shapes signaling networks relevant to tissue-specific physiology.

CYP4F8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CYP4F8 expression without altering the underlying DNA sequence.

CYP4F8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CYP4F8 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 CYP4F8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CYP4F8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CYP4F8 locus and enabling the study of CYP4F8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CYP4F8 pathway restoration in tumor cells with silenced or reduced CYP4F8 expression.

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