NALP6 CRISPR Activation Plasmid (h): sc-404548-ACT

NLRP6 (NALP6) is a cytosolic NOD-like receptor that functions as a pattern-recognition sensor and scaffold for inflammasome assembly, linking microbial and danger signals to innate immune transcriptional programs. It modulates caspase-1 activation, IL-1 family cytokine processing, and downstream NF-κB and MAPK signaling, thereby influencing epithelial barrier integrity and mucosal immune homeostasis. NALP6 activity has been connected to regulation of host–microbiome interactions and inflammatory responses in gastrointestinal tissues, and dysregulation is studied in the context of chronic inflammation and tumor-associated immune microenvironments. As a result, NLRP6 is commonly investigated in pathways controlling pyroptosis, cytokine maturation, and innate immune crosstalk with epithelial and stromal cells.

NALP6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NLRP6 expression without altering the underlying DNA sequence.

NALP6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NLRP6 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 NLRP6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NALP6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NLRP6 locus and enabling the study of NALP6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NALP6 pathway restoration in tumor cells with silenced or reduced NLRP6 expression.

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