NHE-9 CRISPR Activation Plasmid (h): sc-410416-ACT

SLC9A9 encodes the endosomal Na+/H+ exchanger NHE-9, a multi-pass membrane protein that regulates luminal pH and sodium homeostasis in recycling and sorting endosomes. By tuning endosomal acidification, NHE-9 influences receptor trafficking, ligand–receptor dissociation, and the balance between recycling and lysosomal degradation, thereby shaping signaling outputs and membrane protein composition. This function connects SLC9A9 to vesicular transport networks and pH-dependent processes that impact synaptic activity, nutrient transporter regulation, and cellular stress responses. Altered SLC9A9 expression or endosomal pH control has been associated with neurodevelopmental phenotypes and other conditions linked to dysfunctional trafficking and signaling.

NHE-9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC9A9 expression without altering the underlying DNA sequence.

NHE-9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC9A9 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 SLC9A9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NHE-9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC9A9 locus and enabling the study of NHE-9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NHE-9 pathway restoration in tumor cells with silenced or reduced SLC9A9 expression.

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