Cyclophilin F CRISPR Activation Plasmid (h): sc-402902-ACT

PPIF encodes human cyclophilin F, a mitochondrial peptidyl-prolyl cis-trans isomerase that contributes to protein folding and quality control within the mitochondrial matrix and inner membrane environment. Cyclophilin F participates in regulation of mitochondrial permeability transition and influences reactive oxygen species homeostasis, linking it to cellular stress responses, bioenergetics, and apoptotic signaling. Through these functions, PPIF activity is often studied in contexts of mitochondrial dysfunction, where altered permeability transition and redox imbalance are associated with neurodegeneration, ischemia-reperfusion injury models, and metabolic disease mechanisms. PPIF also serves as a node for dissecting how chaperone-like folding enzymes tune mitochondrial proteostasis and signaling under inflammatory or oxidative stress.

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

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

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