NBC4 CRISPR Activation Plasmid (h): sc-412900-ACT

Human SLC4A5 encodes the electroneutral sodium bicarbonate cotransporter NBC4, a membrane transporter that mediates Na⁺-coupled HCO₃⁻ flux to support intracellular pH regulation, bicarbonate homeostasis, and epithelial ion transport. By shaping cytosolic buffering capacity and transepithelial bicarbonate movement, NBC4 contributes to processes such as renal and intestinal electrolyte handling and cellular responses to acid–base stress. Altered SLC4A5 activity has been investigated in the context of dysregulated ion transport and pH-dependent signaling that can influence excitability, metabolism, and tissue homeostasis. These properties make SLC4A5 a useful target for studying solute carrier biology, membrane transport energetics, and pH-linked regulatory pathways.

NBC4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC4A5 expression without altering the underlying DNA sequence.

NBC4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC4A5 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 SLC4A5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NBC4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC4A5 locus and enabling the study of NBC4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NBC4 pathway restoration in tumor cells with silenced or reduced SLC4A5 expression.

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