Arnt 1 CRISPR Activation Plasmid (h): sc-401039-ACT

Human ARNT (aryl hydrocarbon receptor nuclear translocator; HIF-1β) encodes a bHLH-PAS transcription factor that heterodimerizes with partners such as AHR and HIF-1α/HIF-2α to coordinate stimulus-dependent gene expression programs. ARNT-containing complexes regulate xenobiotic response signaling, cellular adaptation to hypoxia, angiogenic and metabolic transcriptional networks, and broader stress-responsive pathways that influence proliferation and differentiation. Through these roles, ARNT is frequently studied in contexts linking environmental exposures to transcriptional rewiring, and in disease-relevant models where hypoxia signaling and AHR pathway activity contribute to altered cell state and gene expression. Its central position in PAS-family transcriptional circuitry makes ARNT a useful node for dissecting pathway cross-talk and transcriptional control mechanisms.

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

Arnt 1 CRISPR Activation Plasmid (h) 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.