HSBP1 CRISPR Activation Plasmid (h): sc-416835-ACT

Human HSBP1 (heat shock factor binding protein 1) is a small coiled-coil regulator of the heat shock response that binds heat shock factor 1 (HSF1) and modulates HSF1 trimerization and transcriptional output. Through this negative feedback control, HSBP1 helps tune proteostasis programs including induction of chaperones such as HSP70/HSP90 and broader stress-adaptation pathways linked to protein folding, translation, and quality control. Altered HSBP1 activity can shift cellular tolerance to proteotoxic stress and intersect with processes relevant to tumor biology, neurodegeneration, and other conditions characterized by dysregulated stress signaling. These roles make HSBP1 a useful target for studying heat shock pathway dynamics, stress resilience, and transcriptional network buffering in human cells.

HSBP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HSBP1 expression without altering the underlying DNA sequence.

HSBP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HSBP1 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 HSBP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HSBP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HSBP1 locus and enabling the study of HSBP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HSBP1 pathway restoration in tumor cells with silenced or reduced HSBP1 expression.

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