NHERF-1 CRISPR Activation Plasmid (h): sc-404885-ACT

SLC9A3R1 encodes NHERF-1 (Na+/H+ exchanger regulatory factor 1), a PDZ-domain scaffold that organizes membrane receptors, ion transporters, and signaling enzymes at the apical plasma membrane. NHERF-1 modulates epithelial ion and fluid transport through interactions with NHE3 (SLC9A3), CFTR, and GPCRs, and links these complexes to the actin cytoskeleton via ERM proteins. Through coordination of receptor trafficking and signal propagation, NHERF-1 influences pathways including cAMP/PKA signaling, PI3K–AKT, and Wnt/β-catenin context-dependent responses. Dysregulated NHERF-1 expression or localization has been associated with altered epithelial homeostasis and signaling phenotypes relevant to cancer biology, renal and intestinal transport regulation, and inflammatory microenvironments.

NHERF-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC9A3R1 expression without altering the underlying DNA sequence.

NHERF-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC9A3R1 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 SLC9A3R1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NHERF-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC9A3R1 locus and enabling the study of NHERF-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NHERF-1 pathway restoration in tumor cells with silenced or reduced SLC9A3R1 expression.

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