BACE2 CRISPR Activation Plasmid (h): sc-403147-ACT

BACE2 (beta-site APP-cleaving enzyme 2) is a type I transmembrane aspartyl protease that participates in regulated intramembrane proteolysis and influences trafficking and turnover of select membrane proteins. In human cells, BACE2 has been linked to proteostasis pathways involving endosomal–lysosomal processing and secretory compartment dynamics, with functional connections to amyloid precursor protein family processing and broader substrate-dependent signaling effects. Its expression and activity have been studied in contexts including neurobiology and metabolic regulation, where altered protease-mediated processing can perturb cellular homeostasis. These attributes make BACE2 a useful target for investigating protease-driven network remodeling and pathway crosstalk in disease-relevant model systems.

BACE2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BACE2 expression without altering the underlying DNA sequence.

BACE2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BACE2 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 BACE2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BACE2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BACE2 locus and enabling the study of BACE2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BACE2 pathway restoration in tumor cells with silenced or reduced BACE2 expression.

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