SLC12A8 CRISPR Activation Plasmid (m): sc-431364-ACT

Slc12a8 encodes SLC12A8, a member of the solute carrier 12 (SLC12) family of cation-coupled transporters that regulates membrane ion gradients and contributes to epithelial and cellular osmotic homeostasis. Through modulation of coupled Na⁺/K⁺/Cl⁻ flux and related electrochemical driving forces, SLC12A8 can influence cell volume control, transepithelial transport processes, and downstream signaling linked to metabolic and stress-responsive pathways. Altered activity of SLC12 transporters is often associated with perturbations in electrolyte balance and tissue physiology, making Slc12a8 a useful target for investigating transporter-dependent mechanisms in normal and disease-relevant contexts. In mouse models, Slc12a8 regulation can be leveraged to study how ion transport interfaces with barrier function, inflammation, and metabolic adaptation.

SLC12A8 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Slc12a8 expression without altering the underlying DNA sequence.

SLC12A8 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Slc12a8 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 Slc12a8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SLC12A8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Slc12a8 locus and enabling the study of SLC12A8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SLC12A8 pathway restoration in tumor cells with silenced or reduced Slc12a8 expression.

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