caspase-1 CRISPR Activation Plasmid (m): sc-419461-ACT

Mouse Casp1 encodes caspase-1, an inflammatory cysteine protease that is activated downstream of canonical inflammasomes such as NLRP3, NLRC4, and AIM2. Upon activation, caspase-1 cleaves pro–IL-1β and pro–IL-18 into their mature cytokines and processes gasdermin D to initiate pyroptotic cell death, linking pathogen sensing and sterile danger signals to inflammatory output. These pathways shape innate immune responses in myeloid and barrier tissues and are widely studied in contexts including infection, autoinflammatory phenotypes, metabolic inflammation, and neuroinflammation. Caspase-1 signaling intersects with NF-κB priming, cytokine networks, and cell death programs, making Casp1 a useful node for dissecting immunometabolic and stress-response mechanisms in mouse models.

caspase-1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Casp1 expression without altering the underlying DNA sequence.

caspase-1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Casp1 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 Casp1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous caspase-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Casp1 locus and enabling the study of caspase-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of caspase-1 pathway restoration in tumor cells with silenced or reduced Casp1 expression.

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