SLC41A1 CRISPR Activation Plasmid (h): sc-406258-ACT

SLC41A1 encodes a plasma membrane magnesium transporter that contributes to cellular Mg2+ efflux and overall magnesium homeostasis, a key determinant of enzyme activity, ATP-dependent metabolism, and ion channel regulation. By shaping intracellular Mg2+ availability, SLC41A1 influences signaling processes coupled to nucleotide-binding proteins and phosphorylation networks, and can affect mitochondrial function and cellular stress responses. Altered regulation of magnesium transport has been associated with dysregulated neuronal excitability and metabolic phenotypes, making SLC41A1 a useful node for studying ion homeostasis in disease-relevant contexts. Its expression and activity are therefore frequently examined in models of neurobiology, bioenergetics, and transport physiology.

SLC41A1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC41A1 expression without altering the underlying DNA sequence.

SLC41A1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC41A1 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 SLC41A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SLC41A1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC41A1 locus and enabling the study of SLC41A1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SLC41A1 pathway restoration in tumor cells with silenced or reduced SLC41A1 expression.

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