PSM CRISPR Activation Plasmid (h): sc-401947-ACT

FOLH1 encodes prostate-specific membrane antigen (PSMA/PSM), a type II transmembrane metallopeptidase with glutamate carboxypeptidase activity that modulates extracellular folate metabolism and glutamatergic signaling. In peripheral tissues, FOLH1 contributes to processing folate polyglutamates, supporting one-carbon metabolism and nucleotide biosynthesis, while in the nervous system it functions as NAALADase to hydrolyze N-acetylaspartylglutamate and influence synaptic glutamate availability. PSMA is also expressed on tumor-associated neovasculature and in prostate lineage contexts, making FOLH1 regulation relevant to studies of angiogenic programs, metabolic reprogramming, and cell-surface enzyme biology in cancer-associated microenvironments. These functions connect FOLH1 to pathways governing nutrient utilization, extracellular peptide processing, and signaling-dependent changes in cell state.

PSM CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FOLH1 expression without altering the underlying DNA sequence.

PSM CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FOLH1 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 FOLH1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PSM expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FOLH1 locus and enabling the study of PSM-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PSM pathway restoration in tumor cells with silenced or reduced FOLH1 expression.

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