ATPIF1 CRISPR Activation Plasmid (h): sc-403038-ACT

ATPIF1 (ATPase inhibitory factor 1) encodes a mitochondrial protein that binds the F1F0-ATP synthase to restrain ATP hydrolysis when mitochondrial membrane potential collapses, thereby helping preserve cellular ATP during energetic stress. By modulating oxidative phosphorylation efficiency and coupling state, ATPIF1 influences mitochondrial bioenergetics, reactive oxygen species balance, and metabolic reprogramming between respiration and glycolysis. Its activity links to mitochondrial quality control and stress-adaptive signaling pathways that shape cell survival and proliferation under hypoxia or nutrient limitation. Dysregulated ATPIF1 expression has been associated with altered mitochondrial function in cancer metabolism and with bioenergetic defects implicated in cardiometabolic and neurodegenerative disease models.

ATPIF1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ATPIF1 expression without altering the underlying DNA sequence.

ATPIF1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ATPIF1 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 ATPIF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ATPIF1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ATPIF1 locus and enabling the study of ATPIF1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ATPIF1 pathway restoration in tumor cells with silenced or reduced ATPIF1 expression.

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