NKp30 CRISPR Activation Plasmid (h): sc-403117-ACT

NCR3 encodes NKp30, an activating receptor expressed on natural killer (NK) cells and subsets of innate-like T cells that contributes to immune surveillance through recognition of stress-induced ligands on target cells. NKp30 engagement triggers ITAM-dependent signaling via associated adaptor molecules, promoting downstream pathways such as SYK/ZAP70 activation, calcium flux, MAPK signaling, cytotoxic granule exocytosis, and cytokine production. Altered NCR3/NKp30 expression or function has been linked to differences in NK-cell responsiveness within tumor microenvironments and during viral infection, and it is frequently studied in the context of immune evasion and inflammation. As a surface receptor central to innate effector activation, NKp30 provides a tractable node for dissecting signaling thresholds and receptor–ligand biology in human immune cells.

NKp30 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NCR3 expression without altering the underlying DNA sequence.

NKp30 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NCR3 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 NCR3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NKp30 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NCR3 locus and enabling the study of NKp30-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NKp30 pathway restoration in tumor cells with silenced or reduced NCR3 expression.

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