Frataxin CRISPR Activation Plasmid (h): sc-401628-ACT

FXN encodes the human mitochondrial protein frataxin, a central regulator of iron–sulfur (Fe–S) cluster biogenesis that supports oxidative phosphorylation, TCA-cycle enzyme function, and redox homeostasis. Frataxin participates in mitochondrial iron handling and coordinates assembly of Fe–S cofactors required for electron transport and multiple metabolic enzymes. Reduced FXN expression is associated with mitochondrial dysfunction, oxidative stress, and altered iron metabolism, linking this pathway to neurodegeneration and cardiometabolic phenotypes. As a result, FXN is widely studied in models that probe mitochondrial quality control, bioenergetic remodeling, and stress-response signaling.

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

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

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