HSP70-2 CRISPR Activation Plasmid (h): sc-417733-ACT

HSPA1B encodes the inducible chaperone HSP70-2, a central component of the cellular heat shock response that preserves proteostasis during proteotoxic stress. HSP70-2 binds exposed hydrophobic regions on nascent or misfolded proteins to promote refolding, prevent aggregation, and facilitate triage decisions between refolding and degradation through the ubiquitin–proteasome system and autophagy. By coordinating with co-chaperones and heat shock factor signaling, HSP70-2 influences apoptosis, inflammatory signaling, and recovery from oxidative, thermal, and ER-associated stress. Altered HSP70 network activity has been associated with phenotypes relevant to tumor cell survival, neurodegeneration, and stress-induced cytotoxicity, making HSPA1B a useful node for mechanistic studies of stress adaptation.

HSP70-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HSPA1B expression without altering the underlying DNA sequence.

HSP70-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HSPA1B 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 HSPA1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HSP70-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HSPA1B locus and enabling the study of HSP70-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HSP70-2 pathway restoration in tumor cells with silenced or reduced HSPA1B expression.

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