ATP5J CRISPR Activation Plasmid (m): sc-419251-ACT

Atp5j encodes ATP5J (also known as F6), a small accessory subunit of the mitochondrial F1Fo ATP synthase (Complex V) that supports assembly and efficient coupling of proton flux to ATP production during oxidative phosphorylation. By modulating mitochondrial bioenergetics, ATP5J contributes to maintenance of cellular ATP/ADP balance, mitochondrial membrane potential, and metabolic flexibility under changing energetic demands. Perturbations in ATP synthase function and downstream respiratory chain activity are linked to mitochondrial dysfunction phenotypes that impact high-energy tissues and intersect with oxidative stress and apoptosis-related processes. In mouse models, altered mitochondrial ATP production is frequently leveraged to study mechanisms relevant to neuromuscular, cardiac, and metabolic disease biology at the level of mitochondrial pathways.

ATP5J CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Atp5j expression without altering the underlying DNA sequence.

ATP5J CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Atp5j 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 Atp5j transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ATP5J expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Atp5j locus and enabling the study of ATP5J-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ATP5J pathway restoration in tumor cells with silenced or reduced Atp5j expression.

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