ARALAR CRISPR Activation Plasmid (h): sc-403673-ACT

SLC25A12 encodes ARALAR, a calcium-regulated mitochondrial aspartate–glutamate carrier (AGC1) that supports the malate–aspartate shuttle and couples cytosolic and mitochondrial redox balance. By exchanging aspartate and glutamate across the inner mitochondrial membrane, ARALAR helps sustain NADH transfer, TCA cycle anaplerosis, and biosynthetic fluxes linked to neuronal energy metabolism and myelin-associated pathways. Its activity integrates mitochondrial function with calcium signaling and amino acid metabolism, influencing cellular responses to energetic stress. Altered SLC25A12 expression or function has been associated with neurodevelopmental and neurometabolic phenotypes and is studied in contexts involving mitochondrial dysfunction and excitatory neurotransmitter handling.

ARALAR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC25A12 expression without altering the underlying DNA sequence.

ARALAR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC25A12 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 SLC25A12 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ARALAR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC25A12 locus and enabling the study of ARALAR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ARALAR pathway restoration in tumor cells with silenced or reduced SLC25A12 expression.

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