NALP9A CRISPR Activation Plasmid (m): sc-433227-ACT

Mouse Nlrp9a encodes NALP9A, a member of the NOD-like receptor (NLR) family implicated in innate immune sensing and inflammasome-related signaling. Like other pyrin domain–containing NLR proteins, NALP9A is associated with pathways that regulate caspase activation, inflammatory cytokine processing, and stress-responsive cell death programs. Nlrp9a expression has been linked to epithelial and reproductive tissues, suggesting roles in barrier-associated immune surveillance and developmental context–dependent inflammation. Dysregulation of NLR-mediated signaling is relevant to studies of autoinflammation and mucosal immune homeostasis, providing a framework for interrogating how Nlrp9a influences inflammatory phenotypes in mouse models.

NALP9A CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Nlrp9a expression without altering the underlying DNA sequence.

NALP9A CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Nlrp9a 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 Nlrp9a transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NALP9A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Nlrp9a locus and enabling the study of NALP9A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NALP9A pathway restoration in tumor cells with silenced or reduced Nlrp9a expression.

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