Cryopyrin/NALP3/NLRP3 CRISPR Activation Plasmid (m): sc-432122-ACT

Mouse Nlrp3 encodes cryopyrin (NALP3/NLRP3), a cytosolic pattern-recognition receptor that nucleates the NLRP3 inflammasome in response to diverse danger signals such as ionic flux, mitochondrial stress, and lysosomal damage. Inflammasome assembly promotes ASC-dependent caspase-1 activation, leading to maturation and release of IL-1β and IL-18 and induction of pyroptotic cell death. NLRP3 signaling integrates innate immune sensing with NF-κB priming and downstream cytokine networks that shape myeloid activation, tissue inflammation, and host defense. Dysregulated NLRP3 activity is widely used as a mechanistic axis for modeling autoinflammatory phenotypes and inflammatory contributions to metabolic, neuroinflammatory, and cardiovascular disease processes in mice.

Cryopyrin/NALP3/NLRP3 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Nlrp3 expression without altering the underlying DNA sequence.

Cryopyrin/NALP3/NLRP3 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Nlrp3 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 Nlrp3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cryopyrin/NALP3/NLRP3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Nlrp3 locus and enabling the study of Cryopyrin/NALP3/NLRP3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cryopyrin/NALP3/NLRP3 pathway restoration in tumor cells with silenced or reduced Nlrp3 expression.

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