Arnt 2 CRISPR Activation Plasmid (h): sc-403101-ACT

ARNT2 (aryl hydrocarbon receptor nuclear translocator 2) encodes a basic helix–loop–helix PAS transcription factor that heterodimerizes with PAS-domain partners to couple environmental and developmental cues to gene expression programs. As a nuclear translocator, ARNT2 participates in oxygen- and xenobiotic-responsive transcriptional networks, integrating signaling that shapes neuronal differentiation, metabolic adaptation, and cellular stress responses. ARNT2-regulated transcription is linked to pathways governing hypoxia-responsive gene expression and neurodevelopmental patterning, with relevance to disorders involving altered neural gene regulation and stress signaling. Dysregulated ARNT2 activity has also been studied in the context of pathway rewiring in cancer and other diseases where bHLH-PAS transcriptional control influences cell state and survival programs.

Arnt 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ARNT2 expression without altering the underlying DNA sequence.

Arnt 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ARNT2 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 ARNT2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Arnt 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ARNT2 locus and enabling the study of Arnt 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Arnt 2 pathway restoration in tumor cells with silenced or reduced ARNT2 expression.

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