HSP 90α CRISPR Activation Plasmid (h): sc-400088-ACT

HSP90AA1 encodes the human heat shock protein HSP90α (HSP 90α), an abundant ATP-dependent molecular chaperone that stabilizes and folds a wide range of client proteins, including kinases, steroid hormone receptors, and transcription factors. HSP90α supports proteostasis and cellular stress adaptation through coordination with co-chaperones, regulation of protein maturation, and buffering of misfolded proteins during heat shock and other stressors. Through its client network, HSP90α interfaces with signaling pathways such as MAPK and PI3K–AKT, as well as cell cycle control and DNA damage response processes. Dysregulated HSP90AA1 expression or dependency is frequently studied in contexts including tumor cell signaling rewiring, neurodegenerative protein aggregation, and inflammatory stress responses, making it a central node for mechanistic pathway research.

HSP 90α CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HSP90AA1 expression without altering the underlying DNA sequence.

HSP 90α CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HSP90AA1 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 HSP90AA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HSP 90α expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HSP90AA1 locus and enabling the study of HSP 90α-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HSP 90α pathway restoration in tumor cells with silenced or reduced HSP90AA1 expression.

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