HSP 20 CRISPR Activation Plasmid (h): sc-403164-ACT

HSPB6 encodes the small heat shock protein HSP 20, a stress-inducible molecular chaperone that modulates protein folding, cytoskeletal dynamics, and proteostasis. In smooth muscle and cardiovascular cell types, HSP 20 participates in signaling programs linked to cAMP/PKA and nitric oxide–cGMP pathways, with phosphorylation-dependent effects on actin remodeling and contractile tone. HSP 20 also interfaces with apoptosis and oxidative stress responses, influencing cell survival under proteotoxic and inflammatory conditions. Dysregulated HSPB6 expression or signaling has been associated with altered vascular reactivity, cardiomyocyte stress adaptation, and smooth muscle dysfunction, making it relevant for mechanistic studies of cardiovascular and airway biology.

HSP 20 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HSPB6 expression without altering the underlying DNA sequence.

HSP 20 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HSPB6 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 HSPB6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HSP 20 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HSPB6 locus and enabling the study of HSP 20-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HSP 20 pathway restoration in tumor cells with silenced or reduced HSPB6 expression.

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