NPT2b CRISPR Activation Plasmid (h): sc-403580-ACT

SLC34A2 encodes the sodium-dependent phosphate transporter NPT2b (NaPi-IIb), a multi-pass membrane protein that mediates inorganic phosphate uptake across epithelial surfaces. NPT2b supports phosphate homeostasis needed for nucleotide synthesis, phospholipid metabolism, and energy-dependent signaling, linking transporter activity to broader metabolic and differentiation programs. In human tissues, SLC34A2 expression is prominent in lung and other epithelia, where regulated phosphate handling influences alveolar and mucosal physiology. Altered SLC34A2 function or expression has been associated with epithelial dysfunction and phosphate-related pathobiology, making it a useful node for studying membrane transport, nutrient sensing, and epithelial remodeling.

NPT2b CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC34A2 expression without altering the underlying DNA sequence.

NPT2b CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC34A2 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 SLC34A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NPT2b expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC34A2 locus and enabling the study of NPT2b-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NPT2b pathway restoration in tumor cells with silenced or reduced SLC34A2 expression.

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