PHT2 CRISPR Activation Plasmid (m): sc-425761-ACT

Mouse Slc15a3 encodes the proton-coupled histidine and oligopeptide transporter PHT2 (SLC15A3), an endolysosomal membrane carrier that supports peptide and histidine flux in acidic intracellular compartments. By regulating substrate availability and luminal pH-dependent transport, PHT2 contributes to lysosome-associated metabolism and peptide handling that can influence antigen processing and innate immune signaling networks. Expression of Slc15a3 is enriched in myeloid lineages and is commonly linked to inflammatory stimulation, positioning it within pathways that intersect with host defense and stress-responsive transcriptional programs. Dysregulated lysosomal transport and immune activation are relevant to models of chronic inflammation and immune-mediated tissue injury, making Slc15a3 a useful node for mechanistic studies.

PHT2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Slc15a3 expression without altering the underlying DNA sequence.

PHT2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Slc15a3 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 Slc15a3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PHT2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Slc15a3 locus and enabling the study of PHT2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PHT2 pathway restoration in tumor cells with silenced or reduced Slc15a3 expression.

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