CYP2A CRISPR Activation Plasmid (h): sc-403450-ACT

CYP2A13 encodes a human cytochrome P450 monooxygenase in the CYP2A subfamily that catalyzes oxidative metabolism of diverse xenobiotics and endogenous substrates. As a component of phase I biotransformation, CYP2A13 supports redox-dependent reactions that shape cellular exposure to inhaled and systemic chemicals, influencing downstream detoxification and oxidative stress pathways. Its expression and activity contribute to inter-individual variability in metabolic capacity, making it relevant to studies of chemical sensitivity and genotype–phenotype relationships in exposure biology. CYP2A13 is therefore widely used as a model for investigating how P450-mediated metabolism intersects with epithelial biology, inflammatory signaling, and carcinogen bioactivation mechanisms in human systems.

CYP2A13 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CYP2A13 expression without altering the underlying DNA sequence.

CYP2A13 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CYP2A13 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 CYP2A13 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CYP2A13 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CYP2A13 locus and enabling the study of CYP2A13-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CYP2A13 pathway restoration in tumor cells with silenced or reduced CYP2A13 expression.

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