HSP 40-4 CRISPR Activation Plasmid (h): sc-403819-ACT

DNAJA1 encodes the human co-chaperone HSP 40-4 (Hsp40/DnaJ homolog), a J-domain protein that stimulates HSPA/Hsp70 ATPase activity to promote client protein folding, refolding, and triage during proteostasis stress. By coordinating with Hsp70 and other chaperone networks, HSP 40-4 contributes to protein quality control, management of misfolded proteins, and cellular responses to heat shock and oxidative stress, with functional links to ubiquitin–proteasome and autophagy pathways. Altered chaperone capacity and proteostasis imbalance are implicated in cancer cell stress tolerance and in protein misfolding pathologies, making DNAJA1 a useful node for investigating stress-adaptation mechanisms and chaperone-dependent regulation of signaling proteins.

HSP 40-4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DNAJA1 expression without altering the underlying DNA sequence.

HSP 40-4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DNAJA1 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 DNAJA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HSP 40-4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DNAJA1 locus and enabling the study of HSP 40-4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HSP 40-4 pathway restoration in tumor cells with silenced or reduced DNAJA1 expression.

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