NHE-3 CRISPR Activation Plasmid (h): sc-401420-ACT

SLC9A3 encodes the apical Na+/H+ exchanger NHE-3, a major regulator of electroneutral sodium absorption and intracellular pH homeostasis in epithelial cells. NHE-3 activity integrates with ion transport networks that coordinate fluid balance, bicarbonate handling, and microvillar function, and is dynamically controlled by trafficking and phosphorylation downstream of cAMP/PKA, PKC, and NHERF/ezrin-associated scaffolding. In the intestine and kidney, NHE-3 helps couple sodium uptake to nutrient and acid–base transport processes, shaping transepithelial NaCl and water movement. Altered SLC9A3 expression or regulation has been linked to epithelial transport dysfunction relevant to diarrhea/constipation phenotypes, inflammatory signaling contexts, and renal salt-handling disturbances.

NHE-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC9A3 expression without altering the underlying DNA sequence.

NHE-3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC9A3 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 SLC9A3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NHE-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC9A3 locus and enabling the study of NHE-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NHE-3 pathway restoration in tumor cells with silenced or reduced SLC9A3 expression.

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