CyPB CRISPR Activation Plasmid (h): sc-401888-ACT

PPIB encodes cyclophilin B (CyPB), an endoplasmic reticulum–localized peptidyl-prolyl cis-trans isomerase that accelerates protein folding and supports secretory pathway proteostasis. CyPB participates in ER quality control and stress responses, including coordination of chaperone networks and modulation of calcium-dependent processes that influence cellular homeostasis. Through its isomerase activity and interactions with folding substrates, PPIB links protein maturation to pathways governing extracellular matrix production, inflammatory signaling, and cell survival decisions. Dysregulated PPIB/CyPB activity or expression has been associated with pathological contexts involving aberrant protein handling, immune activation, and tissue remodeling, making it relevant for mechanistic studies in disease models.

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

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

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