ARA70 CRISPR Activation Plasmid (m): sc-424185-ACT

Mouse Ncoa4 encodes ARA70, a nuclear receptor coactivator that modulates transcriptional programs by bridging ligand-activated receptors with chromatin and the basal transcription machinery. ARA70 influences hormone-responsive gene expression, including pathways linked to androgen receptor signaling and broader nuclear receptor–dependent metabolic and differentiation processes. NCOA4 is also implicated in iron homeostasis through roles associated with ferritin turnover, connecting transcriptional regulation with cellular stress responses. Dysregulated NCOA4/ARA70 activity has been studied in contexts of altered endocrine signaling, proliferation control, and iron metabolism–associated phenotypes relevant to disease biology.

ARA70 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Ncoa4 expression without altering the underlying DNA sequence.

ARA70 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Ncoa4 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 Ncoa4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ARA70 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Ncoa4 locus and enabling the study of ARA70-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ARA70 pathway restoration in tumor cells with silenced or reduced Ncoa4 expression.

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