Presenilin 1 CRISPR Activation Plasmid (h): sc-401227-ACT

PSEN1 encodes presenilin 1, the catalytic core of the γ-secretase complex that mediates intramembrane proteolysis of multiple type I membrane proteins, including NOTCH receptors and amyloid precursor protein (APP). Through regulated cleavage events, presenilin 1 influences Notch signaling, cell fate decisions, and protein trafficking within endosomal and lysosomal compartments. PSEN1 activity also impacts calcium homeostasis and synaptic function via processing-dependent modulation of neuronal signaling pathways. Genetic variation and dysregulated γ-secretase function are strongly associated with neurodegenerative mechanisms relevant to Alzheimer’s disease biology, including altered Aβ peptide generation and downstream cellular stress responses.

Presenilin 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PSEN1 expression without altering the underlying DNA sequence.

Presenilin 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PSEN1 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 PSEN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Presenilin 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PSEN1 locus and enabling the study of Presenilin 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Presenilin 1 pathway restoration in tumor cells with silenced or reduced PSEN1 expression.

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