HSP 40 CRISPR Activation Plasmid (h): sc-402810-ACT

DNAJB1 encodes the human HSP 40 (DnaJ homolog subfamily B member 1), a cytosolic co-chaperone that partners with HSP70 to regulate protein folding, refolding, and triage of misfolded substrates. By stimulating HSP70 ATPase activity, HSP 40 supports proteostasis across stress responses, including heat shock and oxidative stress, and interfaces with pathways governing ubiquitin–proteasome turnover and autophagy. DNAJB1 activity influences cellular responses to proteotoxic stress, with research links to neurodegeneration, viral replication, and oncogenic processes where chaperone networks modulate signaling and protein stability. Dysregulation of chaperone/co-chaperone balance is frequently studied for its impact on aggregation-prone proteins and stress-adaptive transcriptional programs.

HSP 40 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DNAJB1 expression without altering the underlying DNA sequence.

HSP 40 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DNAJB1 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 DNAJB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HSP 40 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DNAJB1 locus and enabling the study of HSP 40-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HSP 40 pathway restoration in tumor cells with silenced or reduced DNAJB1 expression.

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