Prohibitin 2 CRISPR Activation Plasmid (h): sc-401282-ACT

PHB2 encodes prohibitin 2, a conserved scaffold protein enriched in the inner mitochondrial membrane and nucleus that helps maintain mitochondrial cristae architecture, respiratory chain integrity, and mitophagy control. Prohibitin 2 participates in mitochondrial quality control pathways, including OPA1-dependent membrane remodeling and PINK1/Parkin-associated turnover, and can influence cell-cycle and transcriptional programs through interactions with nuclear receptors and chromatin-associated factors. Altered PHB2 expression or localization has been linked to disrupted oxidative phosphorylation, elevated reactive oxygen species signaling, and changes in apoptosis susceptibility. These functions make PHB2 relevant for mechanistic studies of metabolic reprogramming, stress responses, and neurodegeneration- and cancer-associated mitochondrial dysfunction.

Prohibitin 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PHB2 expression without altering the underlying DNA sequence.

Prohibitin 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PHB2 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 PHB2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Prohibitin 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PHB2 locus and enabling the study of Prohibitin 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Prohibitin 2 pathway restoration in tumor cells with silenced or reduced PHB2 expression.

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