FKBP8 CRISPR Activation Plasmid (h): sc-403075-ACT

FKBP8 (FK506 binding protein 8), also known as FKBP38, is a mitochondrial- and ER-associated immunophilin that functions as a peptidyl-prolyl cis–trans isomerase and scaffolding factor coordinating protein folding and signaling. In human cells, FKBP8 modulates apoptosis and mitochondrial homeostasis through interactions with BCL2 family proteins and participates in proteostasis pathways including chaperone networks and selective protein turnover. It has been linked to regulation of mTOR signaling and cellular stress responses that influence survival decisions under metabolic or proteotoxic stress. Dysregulated FKBP8 expression or function has been studied in contexts involving aberrant apoptosis control, mitochondrial dysfunction, and altered stress-adaptive signaling relevant to cancer biology and neurodegeneration research.

FKBP8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FKBP8 expression without altering the underlying DNA sequence.

FKBP8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FKBP8 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 FKBP8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FKBP8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FKBP8 locus and enabling the study of FKBP8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FKBP8 pathway restoration in tumor cells with silenced or reduced FKBP8 expression.

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