IFN-γRα CRISPR Activation Plasmid (h): sc-401191-ACT

IFNGR1 encodes the human interferon-γ receptor alpha chain (IFN-γRα), the ligand-binding subunit that initiates interferon-γ signaling at the cell surface. Upon IFN-γ engagement, IFN-γRα cooperates with IFNGR2 to activate JAK1/JAK2 and downstream STAT1, driving transcriptional programs that regulate antigen presentation, macrophage activation, and antimicrobial and inflammatory responses. This pathway interfaces with IRF/ISG networks and shapes cytokine crosstalk that influences immune surveillance and tissue inflammation. Dysregulated IFNGR1 signaling has been linked to altered host defense and immune-mediated pathology, making it relevant for mechanistic studies of infection biology, inflammation, and immuno-oncology signaling.

IFN-γRα CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IFNGR1 expression without altering the underlying DNA sequence.

IFN-γRα CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IFNGR1 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 IFNGR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IFN-γRα expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IFNGR1 locus and enabling the study of IFN-γRα-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IFN-γRα pathway restoration in tumor cells with silenced or reduced IFNGR1 expression.

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