CYP2W1 CRISPR Activation Plasmid (h): sc-404238-ACT

Human CYP2W1 encodes a cytochrome P450 monooxygenase that catalyzes oxidative metabolism of diverse endogenous substrates and xenobiotics, contributing to cellular redox balance and bioactivation/detoxification chemistry. As an ER-associated heme enzyme, CYP2W1 participates in phase I metabolic processes that intersect with lipid oxidation, reactive oxygen species handling, and downstream conjugation pathways. Its expression is typically low in most adult tissues but is reported to be elevated in certain tumor contexts, making it relevant for studying cancer-associated metabolic reprogramming and atypical P450 expression programs. CYP2W1 activity and regulation are therefore useful for investigating xenobiotic response signaling, metabolic liabilities, and gene–environment interactions in human cell models.

CYP2W1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CYP2W1 expression without altering the underlying DNA sequence.

CYP2W1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CYP2W1 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 CYP2W1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CYP2W1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CYP2W1 locus and enabling the study of CYP2W1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CYP2W1 pathway restoration in tumor cells with silenced or reduced CYP2W1 expression.

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