mitochondrial ferritin CRISPR Activation Plasmid (h): sc-401694-ACT

FTMT encodes mitochondrial ferritin, an intramitochondrial iron-storage protein that sequesters ferrous iron and limits iron-catalyzed reactive oxygen species formation. By buffering labile iron within the mitochondrial matrix, FTMT influences iron homeostasis, heme and iron–sulfur cluster metabolism, and broader mitochondrial redox balance. Altered FTMT expression has been linked to cellular vulnerability under oxidative stress and is studied in contexts of neurodegeneration, anemia-related iron dysregulation, and mitochondrial dysfunction. As a mitochondrial iron gatekeeper, FTMT is frequently investigated for its impact on respiration, metabolic remodeling, and stress-response signaling.

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

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

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