Ah Receptor CRISPR Activation Plasmid (m): sc-419054-ACT

Mouse Ahr encodes the aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor that senses xenobiotics and endogenous metabolites to regulate gene expression programs controlling detoxification, barrier function, and immune modulation. Upon activation, AhR translocates to the nucleus, heterodimerizes with ARNT, and binds xenobiotic response elements to induce targets such as Cyp1a1/Cyp1b1, integrating with oxidative stress, inflammatory signaling, and metabolic pathways. AhR activity influences T cell differentiation, macrophage polarization, epithelial homeostasis, and circadian-associated transcriptional networks. Dysregulated AhR signaling has been linked to altered susceptibility to inflammatory disease, toxicant responses, and tumor microenvironment biology in experimental mouse models.

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

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

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