FKBP25 CRISPR Activation Plasmid (h): sc-410783-ACT

Human FKBP3 encodes FKBP25, a nuclear FK506-binding immunophilin with peptidyl-prolyl cis–trans isomerase activity that supports protein folding and conformational regulation of nuclear complexes. FKBP25 interacts with chromatin-associated factors and contributes to transcriptional control, DNA damage response, and cell-cycle–linked nuclear processes, including modulation of p53-dependent signaling. Through these roles, FKBP3 is studied in contexts of genome stability, epigenetic regulation, and stress-adaptive transcriptional programs. Altered FKBP3/FKBP25 activity and expression have been investigated in proliferative and inflammation-associated disease biology as correlates of dysregulated nuclear signaling and chromatin function.

FKBP25 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FKBP3 expression without altering the underlying DNA sequence.

FKBP25 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FKBP3 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 FKBP3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FKBP25 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FKBP3 locus and enabling the study of FKBP25-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FKBP25 pathway restoration in tumor cells with silenced or reduced FKBP3 expression.

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