NHE-2 CRISPR Activation Plasmid (h): sc-404353-ACT

SLC9A2 encodes the Na+/H+ exchanger NHE-2, a plasma membrane antiporter that regulates intracellular pH, cell volume, and sodium absorption by coupling Na+ influx to H+ efflux. NHE-2 activity supports epithelial ion and fluid transport and contributes to maintenance of mucosal barrier function, linking it to pathways governing transepithelial electrolyte movement and acid–base homeostasis. Through pH-dependent control of enzyme activity, cytoskeletal dynamics, and junctional integrity, NHE-2 influences proliferation and stress responses in polarized cells. Dysregulated Na+/H+ exchange has been associated with gastrointestinal and renal transport abnormalities and inflammation-related phenotypes, making SLC9A2 a relevant target for mechanistic studies of epithelial physiology and disease-associated transport remodeling.

NHE-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC9A2 expression without altering the underlying DNA sequence.

NHE-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC9A2 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 SLC9A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NHE-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC9A2 locus and enabling the study of NHE-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NHE-2 pathway restoration in tumor cells with silenced or reduced SLC9A2 expression.

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