NKp44 CRISPR Activation Plasmid (h): sc-407295-ACT

Human NCR2 encodes NKp44, an activating natural killer (NK) cell receptor expressed on activated NK cells and certain innate lymphoid cells that helps integrate immune surveillance signals at the cell surface. NKp44 engages diverse ligands on stressed, infected, or transformed cells and transduces activation through adapter-dependent signaling, contributing to cytotoxic granule release and cytokine production. This receptor participates in pathways controlling innate immune recognition, immunoreceptor tyrosine-based signaling, and regulation of inflammatory responses. Dysregulated NCR2/NKp44 activity and expression patterns have been associated with altered NK cell function in contexts including viral infection, tumor immunology, and chronic inflammatory disease mechanisms.

NKp44 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NCR2 expression without altering the underlying DNA sequence.

NKp44 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NCR2 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 NCR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NKp44 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NCR2 locus and enabling the study of NKp44-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NKp44 pathway restoration in tumor cells with silenced or reduced NCR2 expression.

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