ARX CRISPR Activation Plasmid (h): sc-402055-ACT

ARX (aristaless related homeobox) encodes a homeobox transcription factor that regulates gene expression programs required for neural progenitor proliferation, interneuron differentiation, and regional patterning during forebrain development. Through sequence-specific DNA binding and interaction with co-regulatory complexes, ARX coordinates transcriptional networks that influence neuronal migration and maturation. Dysregulated ARX activity is linked to neurodevelopmental disorders, including X-linked intellectual disability and epilepsy-associated phenotypes, making it a frequent target for studies of developmental gene regulation. In human cell models, ARX provides a tractable node for probing how transcription factor dosage and timing shape lineage decisions.

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

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

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