PIDD CRISPR Activation Plasmid (h): sc-403439-ACT

PIDD1 encodes the p53-induced death domain protein (PIDD), a cytosolic adaptor that integrates genotoxic and cellular stress signals within apoptotic and inflammatory signaling networks. PIDD participates in assembly of multiprotein complexes such as the PIDDosome, influencing caspase-2 activation, DNA damage responses, and cell fate decisions including apoptosis and cell-cycle checkpoint control. Through its death domain–mediated protein interactions, PIDD can modulate NF-κB–related pathways and stress-responsive transcriptional programs. Dysregulated PIDD1/PIDD signaling has been investigated in the context of altered apoptosis sensitivity and genome stability phenotypes relevant to cancer biology and other disorders of stress signaling.

PIDD CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PIDD1 expression without altering the underlying DNA sequence.

PIDD CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PIDD1 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 PIDD1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PIDD expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PIDD1 locus and enabling the study of PIDD-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PIDD pathway restoration in tumor cells with silenced or reduced PIDD1 expression.

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