PEPT2 CRISPR Activation Plasmid (h): sc-404322-ACT

SLC15A2 encodes PEPT2, a high-affinity proton-coupled oligopeptide transporter in the solute carrier family that mediates uptake and reabsorption of di- and tripeptides as well as peptide-like metabolites. PEPT2 contributes to cellular nutrient sensing and amino acid homeostasis by linking transmembrane H+ gradients to peptide transport, with prominent roles in epithelial transport processes and barrier-associated metabolism. Through its handling of peptide substrates and peptidomimetic compounds, PEPT2 is relevant to pharmacokinetic and toxicology research, particularly in tissues where transporter expression can shape intracellular exposure profiles. Altered SLC15A2 activity has been investigated in contexts of renal and epithelial dysfunction and in studies of transporter-mediated susceptibility to xenobiotic stress.

PEPT2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC15A2 expression without altering the underlying DNA sequence.

PEPT2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC15A2 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 SLC15A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PEPT2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC15A2 locus and enabling the study of PEPT2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PEPT2 pathway restoration in tumor cells with silenced or reduced SLC15A2 expression.

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