PAR4 CRISPR Activation Plasmid (h): sc-401142-ACT

PAWR encodes the pro-apoptotic protein PAR4 (prostate apoptosis response-4), a leucine zipper–containing regulator of cell fate that modulates transcriptional and stress-response networks. PAR4 participates in intrinsic apoptosis and growth control by influencing NF-κB signaling, PKC/AKT survival pathways, and caspase-dependent cell death programs, with additional roles in ER stress and proteostasis. Altered PAWR/PAR4 activity has been associated with dysregulated apoptosis, epithelial and neuronal homeostasis, and context-dependent links to tumor biology and neurodegenerative processes. These functions make PAWR a useful node for studying pathway crosstalk between survival signaling, transcriptional regulation, and programmed cell death.

PAR4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PAWR expression without altering the underlying DNA sequence.

PAR4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PAWR 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 PAWR transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PAR4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PAWR locus and enabling the study of PAR4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PAR4 pathway restoration in tumor cells with silenced or reduced PAWR expression.

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