ZBP1 CRISPR Activation Plasmid (m): sc-425482-ACT

Mouse Zbp1 encodes Z-DNA/RNA-binding protein 1 (ZBP1), an interferon-inducible cytosolic nucleic acid sensor that detects Z-form nucleic acids generated during viral infection or cellular stress. Upon activation, ZBP1 engages RHIM-dependent signaling with RIPK1/RIPK3 to coordinate inflammatory transcriptional programs and cell death pathways, including necroptosis and apoptosis, with downstream effects on innate immune gene expression. This axis intersects with type I interferon signaling, NF-κB activation, and inflammasome-associated responses that shape antiviral defense and tissue inflammation. Dysregulated ZBP1 activity has been implicated in inflammatory and autoinflammatory phenotypes, infection susceptibility, and immune-driven pathology, making it a useful node for mechanistic studies of innate immune sensing and regulated cell death.

ZBP1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Zbp1 expression without altering the underlying DNA sequence.

ZBP1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Zbp1 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 Zbp1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZBP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Zbp1 locus and enabling the study of ZBP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZBP1 pathway restoration in tumor cells with silenced or reduced Zbp1 expression.

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