SLC25A3 CRISPR Activation Plasmid (h): sc-404129-ACT

SLC25A3 encodes a mitochondrial inner membrane solute carrier that transports inorganic phosphate into the matrix, supporting oxidative phosphorylation by supplying substrate for ATP synthase. By coupling phosphate import to mitochondrial membrane potential, SLC25A3 helps regulate cellular bioenergetics, reactive oxygen species balance, and metabolic flexibility in energy-demanding tissues. Altered SLC25A3 function has been linked to mitochondrial dysfunction phenotypes affecting muscle and cardiac physiology, making it relevant for studies of mitochondrial myopathies and cardiometabolic stress responses. This gene is also useful for dissecting how phosphate availability intersects with respiratory chain activity and apoptosis-related mitochondrial signaling.

SLC25A3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC25A3 expression without altering the underlying DNA sequence.

SLC25A3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC25A3 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 SLC25A3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SLC25A3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC25A3 locus and enabling the study of SLC25A3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SLC25A3 pathway restoration in tumor cells with silenced or reduced SLC25A3 expression.

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