IFN-γRβ CRISPR Activation Plasmid (h): sc-402794-ACT

IFNGR2 encodes interferon gamma receptor 2 (IFN-γRβ), the accessory signaling subunit of the IFN-γ receptor complex that partners with IFNGR1 to transmit type II interferon cues. Ligand engagement promotes receptor-associated JAK1/JAK2 activation, STAT1 phosphorylation, and transcriptional programs that shape antigen processing and presentation, macrophage activation, and antimicrobial defenses. Through this pathway, IFNGR2 influences inflammatory signaling cross-talk with IRF and NF-κB networks and contributes to cell-intrinsic responses to immune cytokines. Perturbation of IFN-γ receptor signaling is relevant to studies of immune dysregulation, altered host–pathogen interactions, and mechanisms that modulate tumor-immune surveillance in human model systems.

IFN-γRβ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IFNGR2 expression without altering the underlying DNA sequence.

IFN-γRβ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IFNGR2 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 IFNGR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IFN-γRβ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IFNGR2 locus and enabling the study of IFN-γRβ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IFN-γRβ pathway restoration in tumor cells with silenced or reduced IFNGR2 expression.

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