Arnt 1 CRISPR Activation Plasmid (m): sc-419204-ACT

Arnt (aryl hydrocarbon receptor nuclear translocator), also known as ARNT1/HIF-1β, encodes a bHLH-PAS transcription factor that heterodimerizes with partners such as AHR and HIF-1α to coordinate xenobiotic response and oxygen-sensing gene programs. In mouse cells, ARNT integrates environmental and metabolic cues to regulate transcriptional networks controlling angiogenic signaling, glycolytic metabolism, mitochondrial adaptation, and cellular differentiation. Through the AHR/ARNT axis it modulates detoxification pathways (including cytochrome P450 induction), while in the HIF pathway it supports hypoxia-responsive transcriptional regulation. Dysregulated ARNT-dependent signaling is widely used as a mechanistic entry point for studying inflammation, vascular remodeling, and tumor-associated hypoxic adaptation in preclinical models without implying clinical outcomes.

Arnt 1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Arnt expression without altering the underlying DNA sequence.

Arnt 1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Arnt 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 Arnt transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Arnt 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Arnt locus and enabling the study of Arnt 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Arnt 1 pathway restoration in tumor cells with silenced or reduced Arnt expression.

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