HSF2 CRISPR Activation Plasmid (h): sc-402400-ACT

Human HSF2 (heat shock factor 2) is a sequence-specific transcription factor that coordinates heat shock and proteostasis programs by regulating expression of molecular chaperones and other stress-responsive genes. It participates in cellular pathways controlling protein folding, proteasome function, and adaptation to developmental and environmental stress, with context-dependent interactions with other heat shock factors such as HSF1. HSF2 activity has been linked to differentiation and neurodevelopmental processes, and dysregulation of stress-response transcriptional networks involving HSF2 is studied in conditions characterized by proteotoxic stress, including cancer biology and neurodegeneration models.

HSF2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HSF2 expression without altering the underlying DNA sequence.

HSF2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HSF2 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 HSF2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HSF2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HSF2 locus and enabling the study of HSF2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HSF2 pathway restoration in tumor cells with silenced or reduced HSF2 expression.

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