HSP 27 CRISPR Activation Plasmid (h): sc-400285-ACT

HSPB1 encodes heat shock protein 27 (HSP 27), a small heat shock chaperone that stabilizes unfolded proteins and supports proteostasis during cellular stress. HSP 27 modulates actin cytoskeleton dynamics, regulates apoptotic signaling, and contributes to redox and inflammatory responses through stress-activated kinase networks including MAPK/p38 pathways. Through its phosphorylation-dependent oligomerization and interactions with client proteins, HSP 27 influences cell survival, migration, and stress granule biology. Dysregulated HSPB1 expression or function has been associated with cancer cell stress adaptation, neurodegenerative processes, and cardiometabolic injury models, making it a widely used node for pathway and mechanism studies.

HSP 27 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HSPB1 expression without altering the underlying DNA sequence.

HSP 27 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HSPB1 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 HSPB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HSP 27 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HSPB1 locus and enabling the study of HSP 27-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HSP 27 pathway restoration in tumor cells with silenced or reduced HSPB1 expression.

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