SLC12A9 CRISPR Activation Plasmid (h): sc-408925-ACT

SLC12A9 encodes a member of the solute carrier family 12 of cation–chloride transport-related proteins, implicated in regulating ionic homeostasis across intracellular membranes and shaping osmotic balance, pH, and cell volume responses. Through effects on chloride and alkali cation handling, SLC12A9 may influence membrane potential and signaling pathways coupled to stress adaptation and organellar function. Altered expression or dysregulated ion transport processes are frequently linked to epithelial and neuronal physiology, with broader relevance to disorders where electrolyte balance and cellular volume control are perturbed. As a result, SLC12A9 is of interest for mechanistic studies connecting ion transport networks to cell-state regulation in human systems.

SLC12A9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC12A9 expression without altering the underlying DNA sequence.

SLC12A9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC12A9 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 SLC12A9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SLC12A9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC12A9 locus and enabling the study of SLC12A9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SLC12A9 pathway restoration in tumor cells with silenced or reduced SLC12A9 expression.

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