ANT2 CRISPR Activation Plasmid (h): sc-400680-ACT

SLC25A5 encodes adenine nucleotide translocase 2 (ANT2), a mitochondrial inner membrane carrier that exchanges cytosolic ADP for matrix ATP to couple oxidative phosphorylation with cellular energy demand. By regulating adenine nucleotide flux and mitochondrial membrane potential, ANT2 influences bioenergetic homeostasis, redox balance, and susceptibility to mitochondrial permeability transition–associated stress responses. ANT2 activity is linked to metabolic reprogramming and proliferative states, and altered expression has been studied in the context of tumor metabolism and other disorders with mitochondrial dysfunction. As an X-linked solute carrier, SLC25A5 is also relevant to investigations of tissue-specific energy utilization and compensatory signaling among ANT isoforms.

ANT2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC25A5 expression without altering the underlying DNA sequence.

ANT2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC25A5 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 SLC25A5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ANT2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC25A5 locus and enabling the study of ANT2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ANT2 pathway restoration in tumor cells with silenced or reduced SLC25A5 expression.

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