DnaJB4 CRISPR Activation Plasmid (h): sc-403159-ACT

Human DNAJB4 encodes DnaJB4, a J-domain co-chaperone in the HSP40 family that regulates HSP70 ATPase activity to promote client protein folding, refolding, and triage for degradation. Through participation in proteostasis networks, DnaJB4 contributes to cytosolic quality control and stress-adaptive responses, influencing the handling of misfolded or aggregation-prone proteins. These functions intersect with pathways controlling protein homeostasis, including chaperone-assisted autophagy and proteasome-linked clearance mechanisms. Dysregulation of chaperone systems is relevant to disorders characterized by impaired proteostasis, including neurodegeneration and myopathies, making DNAJB4 a useful node for mechanistic studies of cellular stress and protein quality control.

DnaJB4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DNAJB4 expression without altering the underlying DNA sequence.

DnaJB4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DNAJB4 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 DNAJB4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DnaJB4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DNAJB4 locus and enabling the study of DnaJB4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DnaJB4 pathway restoration in tumor cells with silenced or reduced DNAJB4 expression.

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