NALP7 CRISPR Activation Plasmid (h): sc-403262-ACT

NLRP7 encodes NALP7, a cytosolic NOD-like receptor that participates in innate immune sensing and regulation of inflammasome-associated signaling. Through interactions that influence inflammasome assembly, NALP7 can modulate downstream caspase-1 activity and IL-1 family cytokine processing, linking it to inflammatory and stress-response pathways. NLRP7 expression and sequence variation have been associated with reproductive and placental pathobiology, including recurrent hydatidiform mole and related disorders of early embryonic development. These features make NLRP7 a useful target for dissecting innate immune regulation in trophoblast biology, macrophage-like lineages, and other contexts where inflammasome tuning impacts cellular homeostasis.

NALP7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NLRP7 expression without altering the underlying DNA sequence.

NALP7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NLRP7 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 NLRP7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NALP7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NLRP7 locus and enabling the study of NALP7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NALP7 pathway restoration in tumor cells with silenced or reduced NLRP7 expression.

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