HSP 75 CRISPR Activation Plasmid (h): sc-402640-ACT

TRAP1 (HSP 75) is a mitochondrial HSP90 family chaperone that regulates protein folding and quality control within the mitochondrial matrix, supporting respiratory chain integrity and cellular stress tolerance. By modulating mitochondrial permeability transition, reactive oxygen species homeostasis, and bioenergetic flux, TRAP1 connects chaperone activity to metabolic adaptation and proteostasis pathways. TRAP1-dependent rewiring of mitochondrial function has been linked to altered apoptosis sensitivity, oxidative stress responses, and shifts between oxidative phosphorylation and glycolysis in diverse cellular contexts. Dysregulated TRAP1 expression or activity has been associated with tumor biology, neurodegeneration, and other disorders where mitochondrial dysfunction and proteotoxic stress contribute to pathology.

HSP 75 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TRAP1 expression without altering the underlying DNA sequence.

HSP 75 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TRAP1 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 TRAP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HSP 75 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TRAP1 locus and enabling the study of HSP 75-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HSP 75 pathway restoration in tumor cells with silenced or reduced TRAP1 expression.

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