PBR CRISPR Activation Plasmid (h): sc-401624-ACT

Translocator protein TSPO, also known as the peripheral benzodiazepine receptor (PBR), is an outer mitochondrial membrane protein implicated in cholesterol transport, steroidogenesis, and regulation of mitochondrial bioenergetics. TSPO influences reactive oxygen species homeostasis, mitochondrial permeability transition, and apoptotic signaling, linking it to cellular stress responses and inflammatory programs. In immune and glial cells, TSPO expression correlates with activation state and mitochondrial remodeling, making it a widely used molecular readout in neuroinflammation research. Dysregulated TSPO/PBR activity or expression has been associated with altered mitochondrial function in neurological disorders, cancer-associated metabolic reprogramming, and inflammatory pathologies, supporting mechanistic studies of mitochondrial signaling and cell fate.

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

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

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