NLRC5 CRISPR Activation Plasmid (h): sc-410189-ACT

NLRC5 (NOD-like receptor family CARD domain containing 5) is a cytosolic innate immune regulator best known as a transcriptional activator of MHC class I antigen presentation. It coordinates expression of HLA class I heavy chains, β2-microglobulin, TAP1/2, and immunoproteasome components, thereby linking inflammatory signaling to antigen processing and presentation. Through modulation of interferon-responsive programs and NF-κB–associated pathways, NLRC5 influences immune recognition, cytokine tone, and cellular responses to stress. Dysregulated NLRC5 expression has been associated with altered tumor immunogenicity, susceptibility to viral infection, and autoimmune phenotypes, making it a relevant node for studying immune evasion and antigen presentation defects.

NLRC5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NLRC5 expression without altering the underlying DNA sequence.

NLRC5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NLRC5 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 NLRC5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NLRC5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NLRC5 locus and enabling the study of NLRC5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NLRC5 pathway restoration in tumor cells with silenced or reduced NLRC5 expression.

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